Mardiyyah/TAPT_data_V2_split · Datasets at Hugging Face

PMCID stringclasses 24 values	Title stringclasses 24 values	Sentences stringlengths 2 40.7k
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The mRNAs of MHC class I α-heavy chain and β2 m light chain were detected by RT-qPCR.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	It was shown that decreased expression of HIF-1 and promotion of β2-microglobulin were observed after Endostar treatment.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	In addition, elevated levels of MHC class I α-heavy chain mRNA and protein, as well as downregulation of STAT3 and pSTAT3, were also observed following Endostar treatment.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Endostar inhibited HIF-1 expression in A549 and NCI-H1299 lung cancer cells, upregulated expression of MHC class I α-heavy chain and β2 m light chain, with the upregulation of STAT3 and pSTAT3, suggesting involvement of STAT3 pathway.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	It is important because only in combination with MHC class I on target cells can tumor antigenic peptides be recognized by CD8+ CTLs which destroy target cells.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	However, MHC class I is frequently deficient in cancer cells.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	For the last several decades, lung cancer has long been the most common cancer worldwide and the leading cause of death from cancer with poor survival and high fatality rate of this disease.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The individual risk for development of lung cancer arises from several factors, including smoking and age, radon exposure, asbestos, and other harmful substance exposure, socioeconomic deprivation, earlier diagnosis of a malignant tumor, earlier diagnosis of respiratory conditions, such as chronic obstructive pulmonary disease (COPD), family history, as well as particular rare hereditary disorders, such as Li Fraumeni syndrome and the recently described non-syndromic association with germline EGFR mutation.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Besides, inefficient immunosurveillance, which was caused by immune evasion strategy used by cancer including lung cancer, plays an important role.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Cancer cells can be eliminated by the host immune system because, even though they are self origined, the cancer cells differ from their normal counterparts in terms of their biological behaviors, antigenic characteristics, and biochemical characteristics.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Due to an inefficient DNA damage repair system, tumor-specific neoantigens arise and are expressed in cancer cells.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The neoantigens bind to the human leukocyte antigen (HLA) system class I, which are the major histocompatibility class I (MHC-I) molecules in humans.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The neoantigens binding to HLA-I are presented for CTLs.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	CTLs eliminate cancer cells through two main mechanisms: direct perforin-dependent destruction and secretion of inflammatory cytokines, such as tumor necrosis factor (TNF) and interferon (IFN) alpha (INF-α), which increase tumor immune sensitivity.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Despite the host immune system’s capability）to recognize and destroy cancer cells, the host immune system frequently fails to control cancer growth because cancer cells have acquired the capability to evade recognition and destruction by immune system.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Hypoxia is beneficial for malignant tumors to get rid of immune surveillance.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	In solid tumors, because of uncontrolled growth of cancer cells and disorganized neoangiogenesis, cellular oxygen availability is reduced, leading to intratumoral hypoxic conditions.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	In many types of tumor cells, hypoxia-inducible factor 1 (HIF-1) is highly expressed.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	HIF-1 signaling pathways play an important role in metabolic adaptation to hypoxia stress.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	HIF-1 signaling also induces immune checkpoint molecules and immunosuppressive factors to express, which suppresses innate and adaptive immune systems in order to escape from immune attack.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	MHC class I molecules, which are necessary for antigen-presentation, are also down-regulated by HIF-1, thus limiting T cells recognizing tumor cells.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Human cancers of different histological origins have shown altered or complete loss of MHC class I, which subsequently affects the final outcome of immunotherapy.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Thus, there is a clear need to improve tumor immunogenicity.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The high expression of HIF-1 in tumor cells and its role in MHC-I down-regulation make it an important target in cancer immunotherapy.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Exploring drugs that inhibit HIF-1 and upregulate MHC-I is an important objective.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Endostar may be an important candidate of such drugs.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Endostar is a human recombinant endostatin which is an attractive anti-angiogenesis protein.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	It has been approved by the State Food and Drug Administration of China (CFDA) because of its clinical application on non-small cell lung cancer (NSCLC).
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Nevertheless, besides its anti-angiogenesis, it remains unclear whether Endostar also inhibits HIF-1 expression by upregulating MHC class I expression in cancer cells, which is helpful for immunotherapies, including PD-1/PD-L1 inhibitor, to induce effective responses of cell-mediated immunity against cancers such as lung cancer.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	This study was to demonstrate this effect of Endostar using A549 and NCI-H1299 lung cancer cells.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Human A549 and NCI-H1299 lung cancer cells were maintained in standard cell culture conditions.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The cells whose density were 9 × 10/well were seeded in 6-well culture plates for 24 hours culture to settle.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The temperature was 37 °C.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The atmosphere was humidified and contained 5% CO2 in F-12K (A549 cells) or RPMI-1640 (NCI-H1299 cells) medium supplemented with 10% FBS.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Then, the cells were precultured for 24 h in the medium containing 1% FBS, followed by the addition of Endostar (a Modified Recombinant Human Endostatin expressed and purified in E. coli, which was purchased from Simcere Pharmaceutical Research Co., Ltd., Nanjing, China) into the wells to the indicated final concentration (6.25 μg/ml, 12.5 μg/ml, and 25 μg/ml with 0 μg/ml as control) for a further 24 h incubation.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Cells were collected for analysis.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	All experiments were individually carried out three times.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	This study was approved by the Ethics Committee of the Affiliated Hospital of Chengde Medical University in biomedicine (CYFYLL2021091).
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	To overexpress HIF-1, the cells were transiently transfected with Human HIF1A ORF mammalian expression plasmid (Sino Biological Inc., Beijing, China) using Lipofectamine™ LTX Reagent with PLUS™ Reagent (Invitrogen, CA, USA), according to the manufacturer’s instructions.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Forty-eight hours after transfection, the cells were harvested for assays.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The Endostar treated and untreated cells grown on glass slides were washed and fixed using cold methanol for 5 min.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Next, in order to inhibit the endogenous peroxidase activity, cells were treated with 3% hydrogen peroxide in methanol.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Then, in order to avoid nonspecific binding, cells were blocked with 10% normal serum.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After overnight incubation at 4 °C with goat polyclonal primary antibody against HIF-1 (Affinity biosciences, Suzhou, China) (at a 1:100 dilution), the cells were hybridized by corresponding secondary antibody for 1 h, and finalized with a diaminobenzidine solution to detect the target antigen.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The nucleus of cells was counterstained by hematoxylin before mounting.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	A light microscope was used to examine the slides.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After extraction of the total cellular proteins from the Endostar treated and untreated cells grown in 6-well culture plates by lysis with radioimmunoprecipitation assay (RIPA) buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China) and quantification with BCA Protein Assay Kit (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China), 30 μg of the proteins, diluted in loading buffer and denaturized for 5 minutes at the temperature of 100 °C following keeping on ice for 10 minutes, were subjected to 10% (for HIF1, MHC-1, STAT3, and pSTAT3) or 12% (for β2 m) sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The proteins were then electrotransferred onto polyvinylidene fluoride (PVDF) membrane, blocked for 1 hour with 5% skimmed milk at room temperature, before overnight probe at the temperature of 4 °C using relevant primary antibodies, polyclonal rabbit anti-HIF-1 antibody (Affinity biosciences, Suzhou, China), recombinat monoconal rabbit anti-β2 m antibody (HuaBio Inc., Cambridge, USA), recombinat monoconal rabbit anti-HLA-ABC antibody (HuaBio Inc., Cambridge, USA), recombinat monoconal rabbit anti-STAT3 and anti-pSTAT3 antibody (HuaBio Inc., Cambridge, USA), and mouse monoclonal anti-β-actin antibody (Proteintech, Rosemont, Illinois, USA) following 2-h incubation with horseradish peroxidase-conjugated secondary antibodies.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The protein bands were visualized using enhanced chemiluminescence (ELC) detection system (Amersham, Arlington Heights, IL, USA).
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The expression of β-actin serves as an endogenous control.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The acquired images were quantified with normalizative to β-actin using ImageJ software (Rasband, W.S., ImageJ, U.S. National Institutes of Health, Bethesda, Maryland, USA).
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Total RNAs in Endostar treated and untreated cells grown in 6-well culture were extracted by using Superbrilliant® 6 min High-quality RNA Extraction Kit (Zhongshi Gene Technology, Tianjin, China) under Rnase free condition according to the protocol of manufacturer.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Fast Quant RTkit (Tiangen Biotech Co., Ltd., Beijing, China) was utilized to synthesize cDNAs.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Quantitative real-time PCR (qPCR) was conducted on Roche Cobas z 480 Real-Time PCR Detection System (Roche, Basel, Switzerland) using SuperReal PreMix Plus kit (Tiangen Biotech Co., Ltd., Beijing, China), and the responsible primers, β2 m (Invitrogen, Carlsbad, USA), HLA-ABC α heavy chain (Invitrogen, Carlsbad, USA), and β-actin (Thermo Fisher Scientific Inc., Waltham, Massachusetts, USA), were used.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The expression of β-actin mRNA as a control was used for normalization of the mRNAs expression with 2 method to calculate the fold changes of the mRNAs.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The data were described as mean ± standard deviation (SD) and the statistical significance was determined by one-way ANOVA before LSD test for multiple comparisons and independent-sample t-test for comparison between the two groups.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The level of p < .05 was used to confirm that the differences were statistically significant.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	It was shown with western blot assays that, in A549 lung cancer cells, treatment with Endostar for 24 h decreased the levels of HIF-1 protein in the groups treated with 25 μg/ml Endostar with statistical significance compared with the control group (p = .015), 6.25 μg/ml group (p = .014) and 12.5 μg/ml group (p = .005), and in NCI-H1299 lung cancer cells, treatment with Endostar decreased the levels of HIF-1 protein in the groups treated with 25 μg/ml Endostar with statistical significance compared with the control group (p = .004) and 12.5 μg/ml group (p = .047) (Figure 1 in detail), which was similar to immunocytochemistry showing the reduction of HIF-1 protein in Endostar treated A549 and NCI-H1299 lung cancer cells compared with the control cells (Figure 2).
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Figure 1.Inhibition by Endostar on HIF-1 in A549 and NCI-H1299 lung cancer cells measured by Western blot assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After treatment with Endostar for 24 h, the HIF-1 in A549 and NCI-H1299 lung cancer cells was measured by Western blot assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Error bars indicate the SD.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Asterisks indicate p < .05, significantly different compared with the control; Hashtags indicate p < .05, significantly different compared with the group treated with 6.25 μg/ml Endostar; ampersands indicate p < .05, significantly different compared with the group treated with 12.5 μg/ml Endostar; one-way analysis of variance followed by LSD test.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	A bar graph (labeled with “HIF-1 in A549 cells”) with four bars, the means of the arbitrary units of 1.03, 1.03, 1.06, and 0.88 with asterisk, hashtag, and ampersand being over each bar, which represent the control group, the 6.25µg/ml Endostar group, the 12.5µg/ml Endostar group, and the 25µg/ml Endostar group, respectively, displays the HIF-1 protein expression levels of three independent experiments after treatment of A549 lung cancer cells with Endostar for 24h; a bar graph (labeled with “HIF-1 in NCI-H1299 cells”) with four bars, the means of the arbitrary units of 1.34, 1.00, 1.05, and 0.64 with asterisk and ampersand being over each bar, which represent the control group, the 6.25µg/ml Endostar group, the 12.5µg/ml Endostar group, and the 25µg/ml Endostar group, respectively, displays the HIF-1 protein expression levels of three independent experiments after treatment of NCI-H1299 lung cancer cells with Endostar for 24 h. Corresponding protein bands with Western blot were representatively shown under each bar graph.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Figure 2.Inhibition by Endostar on HIF-1 in A549 and NCI-H1299 lung cancer cells measured by immunocytochemistry.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After treatment with Endostar for 24 h, the HIF-1 in A549 and NCI-H1299 lung cancer cells was measured by immunocytochemistry.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Photographs were taken under microscope (400× magnification).A picture (entitled A549 Cells) with four photographs of A549 lung cancer cells taken under microscope after immunocytochemistry, the Endostar concentration of control, 6.25µg/ml, 12.5µg/ml, and 25µg/ml being under each photograph, respectively.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The intensity of tan color in the cytoplasm, which is lighter and lighter with the increase of the Endostar concentration, displays the levels of HIF-1 after treatment with Endostar for 24 h; a picture (entitled NCI-H1299 Cells) with four photographs of NCI-H1299 lung cancer cells taken under microscope after immunocytochemistry, the Endostar concentration of control, 6.25µg/ml, 12.5µg/ml, and 25µg/ml being under each photograph, respectively.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The intensity of tan color in the cytoplasm, which is lighter and lighter with the increase of the Endostar concentration, displays the levels of HIF-1 after treatment with Endostar for 24 h. Inhibition by Endostar on HIF-1 in A549 and NCI-H1299 lung cancer cells measured by Western blot assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After treatment with Endostar for 24 h, the HIF-1 in A549 and NCI-H1299 lung cancer cells was measured by Western blot assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Error bars indicate the SD.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Asterisks indicate p < .05, significantly different compared with the control; Hashtags indicate p < .05, significantly different compared with the group treated with 6.25 μg/ml Endostar; ampersands indicate p < .05, significantly different compared with the group treated with 12.5 μg/ml Endostar; one-way analysis of variance followed by LSD test.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Inhibition by Endostar on HIF-1 in A549 and NCI-H1299 lung cancer cells measured by immunocytochemistry.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After treatment with Endostar for 24 h, the HIF-1 in A549 and NCI-H1299 lung cancer cells was measured by immunocytochemistry.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Photographs were taken under microscope (400× magnification).
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Beta 2-microglobulin (β2 m) is the invariable light chain of the MHC class I heterodimer molecules and associated with MHC class I down-regulation which is possibly caused by high level of HIF-1.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	It was shown in this study that with the reduction of the HIF-1 level, unlike the β2 m mRNA levels without statistically significant differences among the Endostar treated and untreated groups (p = .098) according to RT-qPCR assay (Figure 3 in detail), the higher relative levels of β2 m protein were found in the A549 cells in the group which were treated with 25 μg/ml Endostar with statistical significance compared with the control group, 6.25 μg/ml group, and 12.5 μg/ml group (p = .017, p = .003, p = .026, respectively), indicating that the effect of Endostar on the β2 m expression in A549 cells may be mRNA independent, and in the NCI-H1299 cells in the groups which were treated with 6.25 μg/ml, 12.5 μg/ml, and 25 μg/ml (p = .000, p = .000, p = .000, respectively) Endostar, with statistical significance compared with the control group (Figure 4 in detail).
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Figure 3.Effects of Endostar on β2-microglobulin mRNA in A549 lung cancer cells measured by RT-qPCR assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After treatment with Endostar for 24 h, the β2-microglobulin mRNA in A549 lung cancer cells was measured by RT-qPCR assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Error bars indicate the SD.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	No significant differences among the groups were found (p > .05); one-way analysis of variance.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	A bar graph (labeled with “β2m mRNA in A549 cells”) with four bars, the means of 2−ΔΔCt values of 1.00, 1.01, 0.92, and 0.98 being over each bar, which represent the control group, the 6.25µg/ml Endostar group, the 12.5µg/ml Endostar group, and the 25µg/ml Endostar group, respectively, displays the β2-microglobulin mRNA levels of three independent experiments after treatment of A549 lung cancer cells with Endostar for 24 h. Figure 4.Upregulation by Endostar on β2-microglobulin protein in A549 and NCI-H1299 lung cancer cells measured by Western blot assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After treatment with Endostar for 24 h, the β2-microglobulin protein in A549 and NCI-H1299 lung cancer cells was measured by Western blot assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Error bars indicate the SD.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Asterisks indicate p < .05, significantly different compared with the control; Hashtags indicate p < .05, significantly different compared with the group treated with 6.25 μg/ml Endostar; ampersands indicate p < .05, significantly different compared with the group treated with 12.5 μg/ml Endostar; one-way analysis of variance followed by LSD test.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	A bar graph (labeled with “β2m in A549 cells”) with four bars, the means of the arbitrary units of 0.96, 0.86, 0.98, and 1.19 with asterisk, hashtag, and ampersand being over each bar, which represent the control group, the 6.25µg/ml Endostar group, the 12.5µg/ml Endostar group, and the 25µg/ml Endostar group, respectively, displays the β2-microglobulin protein expression levels of three independent experiments after treatment of A549 lung cancer cells with Endostar for 24 h; a bar graph (labeled with “β2m in NCI-H1299 cells”) with four bars, the means of the arbitrary units of 0.68, 1.08 with asterisk, 1.05 with asterisk, and 1.18 with asterisk being over each bar, which represent the control group, the 6.25µg/ml Endostar group, the 12.5µg/ml Endostar group, and the 25µg/ml Endostar group, respectively, displays the β2-microglobulin protein expression levels of three independent experiments after treatment of NCI-H1299 lung cancer cells with Endostar for 24 h. Corresponding protein bands with Western blot were representatively shown under each bar graph.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Effects of Endostar on β2-microglobulin mRNA in A549 lung cancer cells measured by RT-qPCR assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After treatment with Endostar for 24 h, the β2-microglobulin mRNA in A549 lung cancer cells was measured by RT-qPCR assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Error bars indicate the SD.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	No significant differences among the groups were found (p > .05); one-way analysis of variance.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Upregulation by Endostar on β2-microglobulin protein in A549 and NCI-H1299 lung cancer cells measured by Western blot assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After treatment with Endostar for 24 h, the β2-microglobulin protein in A549 and NCI-H1299 lung cancer cells was measured by Western blot assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Error bars indicate the SD.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Asterisks indicate p < .05, significantly different compared with the control; Hashtags indicate p < .05, significantly different compared with the group treated with 6.25 μg/ml Endostar; ampersands indicate p < .05, significantly different compared with the group treated with 12.5 μg/ml Endostar; one-way analysis of variance followed by LSD test.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The loss of MHC class I expression was probably caused by high level of HIF-1.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After a treatment of A549 lung cancer cells with Endostar for 24 hours, the relative levels of MHC-I α heavy chain mRNA demonstrated a slight enhancement without statistical significance (p = .840) in the groups treated with 6.25 μg/ml, 12.5 μg/ml, and 25 μg/ml Endostar compared with the control group (Figure 5 in detail).
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	The relative levels of MHC-I protein α heavy chain in the A549 cells in the groups treated with 6.25 μg/ml, 12.5 μg/ml, and 25 μg/ml Endostar were statistically enhanced (p = .048, p = .007, p = .002, respectively) compared with the control group, and in the NCI-H1299 cells in the groups treated with 6.25 μg/ml, 12.5 μg/ml and 25 μg/ml Endostar were statistically enhanced as well (p = .008, p = .007, p = .005, respectively) compared with the control group (Figure 6 in detail).
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Figure 5.Effects of Endostar on the mRNA of MHC class I α-heavy chain in A549 lung cancer cells measured by RT-qPCR assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	After treatment with Endostar for 24 h, the mRNA of MHC class I α-heavy chain in A549 lung cancer cells was measured by RT-qPCR assay.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Error bars indicate the SD.
PMC12101583	Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.	Slight enhancement without statistical significance were found among the groups with Endostar treatment (p > .05); one-way analysis of variance.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The mRNAs of MHC class I α-heavy chain and β2 m light chain were detected by RT-qPCR.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

It was shown that decreased expression of HIF-1 and promotion of β2-microglobulin were observed after Endostar treatment.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

In addition, elevated levels of MHC class I α-heavy chain mRNA and protein, as well as downregulation of STAT3 and pSTAT3, were also observed following Endostar treatment.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Endostar inhibited HIF-1 expression in A549 and NCI-H1299 lung cancer cells, upregulated expression of MHC class I α-heavy chain and β2 m light chain, with the upregulation of STAT3 and pSTAT3, suggesting involvement of STAT3 pathway.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

It is important because only in combination with MHC class I on target cells can tumor antigenic peptides be recognized by CD8+ CTLs which destroy target cells.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

However, MHC class I is frequently deficient in cancer cells.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

For the last several decades, lung cancer has long been the most common cancer worldwide and the leading cause of death from cancer with poor survival and high fatality rate of this disease.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The individual risk for development of lung cancer arises from several factors, including smoking and age, radon exposure, asbestos, and other harmful substance exposure, socioeconomic deprivation, earlier diagnosis of a malignant tumor, earlier diagnosis of respiratory conditions, such as chronic obstructive pulmonary disease (COPD), family history, as well as particular rare hereditary disorders, such as Li Fraumeni syndrome and the recently described non-syndromic association with germline EGFR mutation.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Besides, inefficient immunosurveillance, which was caused by immune evasion strategy used by cancer including lung cancer, plays an important role.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Cancer cells can be eliminated by the host immune system because, even though they are self origined, the cancer cells differ from their normal counterparts in terms of their biological behaviors, antigenic characteristics, and biochemical characteristics.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Due to an inefficient DNA damage repair system, tumor-specific neoantigens arise and are expressed in cancer cells.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The neoantigens bind to the human leukocyte antigen (HLA) system class I, which are the major histocompatibility class I (MHC-I) molecules in humans.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The neoantigens binding to HLA-I are presented for CTLs.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

CTLs eliminate cancer cells through two main mechanisms: direct perforin-dependent destruction and secretion of inflammatory cytokines, such as tumor necrosis factor (TNF) and interferon (IFN) alpha (INF-α), which increase tumor immune sensitivity.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Despite the host immune system’s capability）to recognize and destroy cancer cells, the host immune system frequently fails to control cancer growth because cancer cells have acquired the capability to evade recognition and destruction by immune system.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Hypoxia is beneficial for malignant tumors to get rid of immune surveillance.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

In solid tumors, because of uncontrolled growth of cancer cells and disorganized neoangiogenesis, cellular oxygen availability is reduced, leading to intratumoral hypoxic conditions.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

In many types of tumor cells, hypoxia-inducible factor 1 (HIF-1) is highly expressed.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

HIF-1 signaling pathways play an important role in metabolic adaptation to hypoxia stress.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

HIF-1 signaling also induces immune checkpoint molecules and immunosuppressive factors to express, which suppresses innate and adaptive immune systems in order to escape from immune attack.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

MHC class I molecules, which are necessary for antigen-presentation, are also down-regulated by HIF-1, thus limiting T cells recognizing tumor cells.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Human cancers of different histological origins have shown altered or complete loss of MHC class I, which subsequently affects the final outcome of immunotherapy.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Thus, there is a clear need to improve tumor immunogenicity.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The high expression of HIF-1 in tumor cells and its role in MHC-I down-regulation make it an important target in cancer immunotherapy.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Exploring drugs that inhibit HIF-1 and upregulate MHC-I is an important objective.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Endostar may be an important candidate of such drugs.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Endostar is a human recombinant endostatin which is an attractive anti-angiogenesis protein.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

It has been approved by the State Food and Drug Administration of China (CFDA) because of its clinical application on non-small cell lung cancer (NSCLC).

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Nevertheless, besides its anti-angiogenesis, it remains unclear whether Endostar also inhibits HIF-1 expression by upregulating MHC class I expression in cancer cells, which is helpful for immunotherapies, including PD-1/PD-L1 inhibitor, to induce effective responses of cell-mediated immunity against cancers such as lung cancer.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

This study was to demonstrate this effect of Endostar using A549 and NCI-H1299 lung cancer cells.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Human A549 and NCI-H1299 lung cancer cells were maintained in standard cell culture conditions.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The cells whose density were 9 × 10/well were seeded in 6-well culture plates for 24 hours culture to settle.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The temperature was 37 °C.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The atmosphere was humidified and contained 5% CO2 in F-12K (A549 cells) or RPMI-1640 (NCI-H1299 cells) medium supplemented with 10% FBS.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Then, the cells were precultured for 24 h in the medium containing 1% FBS, followed by the addition of Endostar (a Modified Recombinant Human Endostatin expressed and purified in E. coli, which was purchased from Simcere Pharmaceutical Research Co., Ltd., Nanjing, China) into the wells to the indicated final concentration (6.25 μg/ml, 12.5 μg/ml, and 25 μg/ml with 0 μg/ml as control) for a further 24 h incubation.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Cells were collected for analysis.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

All experiments were individually carried out three times.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

This study was approved by the Ethics Committee of the Affiliated Hospital of Chengde Medical University in biomedicine (CYFYLL2021091).

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

To overexpress HIF-1, the cells were transiently transfected with Human HIF1A ORF mammalian expression plasmid (Sino Biological Inc., Beijing, China) using Lipofectamine™ LTX Reagent with PLUS™ Reagent (Invitrogen, CA, USA), according to the manufacturer’s instructions.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Forty-eight hours after transfection, the cells were harvested for assays.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The Endostar treated and untreated cells grown on glass slides were washed and fixed using cold methanol for 5 min.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Next, in order to inhibit the endogenous peroxidase activity, cells were treated with 3% hydrogen peroxide in methanol.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Then, in order to avoid nonspecific binding, cells were blocked with 10% normal serum.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After overnight incubation at 4 °C with goat polyclonal primary antibody against HIF-1 (Affinity biosciences, Suzhou, China) (at a 1:100 dilution), the cells were hybridized by corresponding secondary antibody for 1 h, and finalized with a diaminobenzidine solution to detect the target antigen.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The nucleus of cells was counterstained by hematoxylin before mounting.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

A light microscope was used to examine the slides.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After extraction of the total cellular proteins from the Endostar treated and untreated cells grown in 6-well culture plates by lysis with radioimmunoprecipitation assay (RIPA) buffer (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China) and quantification with BCA Protein Assay Kit (Beijing Solarbio Science & Technology Co., Ltd., Beijing, China), 30 μg of the proteins, diluted in loading buffer and denaturized for 5 minutes at the temperature of 100 °C following keeping on ice for 10 minutes, were subjected to 10% (for HIF1, MHC-1, STAT3, and pSTAT3) or 12% (for β2 m) sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The proteins were then electrotransferred onto polyvinylidene fluoride (PVDF) membrane, blocked for 1 hour with 5% skimmed milk at room temperature, before overnight probe at the temperature of 4 °C using relevant primary antibodies, polyclonal rabbit anti-HIF-1 antibody (Affinity biosciences, Suzhou, China), recombinat monoconal rabbit anti-β2 m antibody (HuaBio Inc., Cambridge, USA), recombinat monoconal rabbit anti-HLA-ABC antibody (HuaBio Inc., Cambridge, USA), recombinat monoconal rabbit anti-STAT3 and anti-pSTAT3 antibody (HuaBio Inc., Cambridge, USA), and mouse monoclonal anti-β-actin antibody (Proteintech, Rosemont, Illinois, USA) following 2-h incubation with horseradish peroxidase-conjugated secondary antibodies.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The protein bands were visualized using enhanced chemiluminescence (ELC) detection system (Amersham, Arlington Heights, IL, USA).

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The expression of β-actin serves as an endogenous control.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The acquired images were quantified with normalizative to β-actin using ImageJ software (Rasband, W.S., ImageJ, U.S. National Institutes of Health, Bethesda, Maryland, USA).

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Total RNAs in Endostar treated and untreated cells grown in 6-well culture were extracted by using Superbrilliant® 6 min High-quality RNA Extraction Kit (Zhongshi Gene Technology, Tianjin, China) under Rnase free condition according to the protocol of manufacturer.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Fast Quant RTkit (Tiangen Biotech Co., Ltd., Beijing, China) was utilized to synthesize cDNAs.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Quantitative real-time PCR (qPCR) was conducted on Roche Cobas z 480 Real-Time PCR Detection System (Roche, Basel, Switzerland) using SuperReal PreMix Plus kit (Tiangen Biotech Co., Ltd., Beijing, China), and the responsible primers, β2 m (Invitrogen, Carlsbad, USA), HLA-ABC α heavy chain (Invitrogen, Carlsbad, USA), and β-actin (Thermo Fisher Scientific Inc., Waltham, Massachusetts, USA), were used.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The expression of β-actin mRNA as a control was used for normalization of the mRNAs expression with 2 method to calculate the fold changes of the mRNAs.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The data were described as mean ± standard deviation (SD) and the statistical significance was determined by one-way ANOVA before LSD test for multiple comparisons and independent-sample t-test for comparison between the two groups.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The level of p < .05 was used to confirm that the differences were statistically significant.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

It was shown with western blot assays that, in A549 lung cancer cells, treatment with Endostar for 24 h decreased the levels of HIF-1 protein in the groups treated with 25 μg/ml Endostar with statistical significance compared with the control group (p = .015), 6.25 μg/ml group (p = .014) and 12.5 μg/ml group (p = .005), and in NCI-H1299 lung cancer cells, treatment with Endostar decreased the levels of HIF-1 protein in the groups treated with 25 μg/ml Endostar with statistical significance compared with the control group (p = .004) and 12.5 μg/ml group (p = .047) (Figure 1 in detail), which was similar to immunocytochemistry showing the reduction of HIF-1 protein in Endostar treated A549 and NCI-H1299 lung cancer cells compared with the control cells (Figure 2).

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Figure 1.Inhibition by Endostar on HIF-1 in A549 and NCI-H1299 lung cancer cells measured by Western blot assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After treatment with Endostar for 24 h, the HIF-1 in A549 and NCI-H1299 lung cancer cells was measured by Western blot assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Error bars indicate the SD.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Asterisks indicate p < .05, significantly different compared with the control; Hashtags indicate p < .05, significantly different compared with the group treated with 6.25 μg/ml Endostar; ampersands indicate p < .05, significantly different compared with the group treated with 12.5 μg/ml Endostar; one-way analysis of variance followed by LSD test.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

A bar graph (labeled with “HIF-1 in A549 cells”) with four bars, the means of the arbitrary units of 1.03, 1.03, 1.06, and 0.88 with asterisk, hashtag, and ampersand being over each bar, which represent the control group, the 6.25µg/ml Endostar group, the 12.5µg/ml Endostar group, and the 25µg/ml Endostar group, respectively, displays the HIF-1 protein expression levels of three independent experiments after treatment of A549 lung cancer cells with Endostar for 24h; a bar graph (labeled with “HIF-1 in NCI-H1299 cells”) with four bars, the means of the arbitrary units of 1.34, 1.00, 1.05, and 0.64 with asterisk and ampersand being over each bar, which represent the control group, the 6.25µg/ml Endostar group, the 12.5µg/ml Endostar group, and the 25µg/ml Endostar group, respectively, displays the HIF-1 protein expression levels of three independent experiments after treatment of NCI-H1299 lung cancer cells with Endostar for 24 h. Corresponding protein bands with Western blot were representatively shown under each bar graph.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Figure 2.Inhibition by Endostar on HIF-1 in A549 and NCI-H1299 lung cancer cells measured by immunocytochemistry.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After treatment with Endostar for 24 h, the HIF-1 in A549 and NCI-H1299 lung cancer cells was measured by immunocytochemistry.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Photographs were taken under microscope (400× magnification).A picture (entitled A549 Cells) with four photographs of A549 lung cancer cells taken under microscope after immunocytochemistry, the Endostar concentration of control, 6.25µg/ml, 12.5µg/ml, and 25µg/ml being under each photograph, respectively.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The intensity of tan color in the cytoplasm, which is lighter and lighter with the increase of the Endostar concentration, displays the levels of HIF-1 after treatment with Endostar for 24 h; a picture (entitled NCI-H1299 Cells) with four photographs of NCI-H1299 lung cancer cells taken under microscope after immunocytochemistry, the Endostar concentration of control, 6.25µg/ml, 12.5µg/ml, and 25µg/ml being under each photograph, respectively.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The intensity of tan color in the cytoplasm, which is lighter and lighter with the increase of the Endostar concentration, displays the levels of HIF-1 after treatment with Endostar for 24 h. Inhibition by Endostar on HIF-1 in A549 and NCI-H1299 lung cancer cells measured by Western blot assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After treatment with Endostar for 24 h, the HIF-1 in A549 and NCI-H1299 lung cancer cells was measured by Western blot assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Error bars indicate the SD.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Asterisks indicate p < .05, significantly different compared with the control; Hashtags indicate p < .05, significantly different compared with the group treated with 6.25 μg/ml Endostar; ampersands indicate p < .05, significantly different compared with the group treated with 12.5 μg/ml Endostar; one-way analysis of variance followed by LSD test.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Inhibition by Endostar on HIF-1 in A549 and NCI-H1299 lung cancer cells measured by immunocytochemistry.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After treatment with Endostar for 24 h, the HIF-1 in A549 and NCI-H1299 lung cancer cells was measured by immunocytochemistry.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Photographs were taken under microscope (400× magnification).

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Beta 2-microglobulin (β2 m) is the invariable light chain of the MHC class I heterodimer molecules and associated with MHC class I down-regulation which is possibly caused by high level of HIF-1.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

It was shown in this study that with the reduction of the HIF-1 level, unlike the β2 m mRNA levels without statistically significant differences among the Endostar treated and untreated groups (p = .098) according to RT-qPCR assay (Figure 3 in detail), the higher relative levels of β2 m protein were found in the A549 cells in the group which were treated with 25 μg/ml Endostar with statistical significance compared with the control group, 6.25 μg/ml group, and 12.5 μg/ml group (p = .017, p = .003, p = .026, respectively), indicating that the effect of Endostar on the β2 m expression in A549 cells may be mRNA independent, and in the NCI-H1299 cells in the groups which were treated with 6.25 μg/ml, 12.5 μg/ml, and 25 μg/ml (p = .000, p = .000, p = .000, respectively) Endostar, with statistical significance compared with the control group (Figure 4 in detail).

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Figure 3.Effects of Endostar on β2-microglobulin mRNA in A549 lung cancer cells measured by RT-qPCR assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After treatment with Endostar for 24 h, the β2-microglobulin mRNA in A549 lung cancer cells was measured by RT-qPCR assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Error bars indicate the SD.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

No significant differences among the groups were found (p > .05); one-way analysis of variance.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

A bar graph (labeled with “β2m mRNA in A549 cells”) with four bars, the means of 2−ΔΔCt values of 1.00, 1.01, 0.92, and 0.98 being over each bar, which represent the control group, the 6.25µg/ml Endostar group, the 12.5µg/ml Endostar group, and the 25µg/ml Endostar group, respectively, displays the β2-microglobulin mRNA levels of three independent experiments after treatment of A549 lung cancer cells with Endostar for 24 h. Figure 4.Upregulation by Endostar on β2-microglobulin protein in A549 and NCI-H1299 lung cancer cells measured by Western blot assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After treatment with Endostar for 24 h, the β2-microglobulin protein in A549 and NCI-H1299 lung cancer cells was measured by Western blot assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Error bars indicate the SD.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Asterisks indicate p < .05, significantly different compared with the control; Hashtags indicate p < .05, significantly different compared with the group treated with 6.25 μg/ml Endostar; ampersands indicate p < .05, significantly different compared with the group treated with 12.5 μg/ml Endostar; one-way analysis of variance followed by LSD test.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

A bar graph (labeled with “β2m in A549 cells”) with four bars, the means of the arbitrary units of 0.96, 0.86, 0.98, and 1.19 with asterisk, hashtag, and ampersand being over each bar, which represent the control group, the 6.25µg/ml Endostar group, the 12.5µg/ml Endostar group, and the 25µg/ml Endostar group, respectively, displays the β2-microglobulin protein expression levels of three independent experiments after treatment of A549 lung cancer cells with Endostar for 24 h; a bar graph (labeled with “β2m in NCI-H1299 cells”) with four bars, the means of the arbitrary units of 0.68, 1.08 with asterisk, 1.05 with asterisk, and 1.18 with asterisk being over each bar, which represent the control group, the 6.25µg/ml Endostar group, the 12.5µg/ml Endostar group, and the 25µg/ml Endostar group, respectively, displays the β2-microglobulin protein expression levels of three independent experiments after treatment of NCI-H1299 lung cancer cells with Endostar for 24 h. Corresponding protein bands with Western blot were representatively shown under each bar graph.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Effects of Endostar on β2-microglobulin mRNA in A549 lung cancer cells measured by RT-qPCR assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After treatment with Endostar for 24 h, the β2-microglobulin mRNA in A549 lung cancer cells was measured by RT-qPCR assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Error bars indicate the SD.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

No significant differences among the groups were found (p > .05); one-way analysis of variance.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Upregulation by Endostar on β2-microglobulin protein in A549 and NCI-H1299 lung cancer cells measured by Western blot assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After treatment with Endostar for 24 h, the β2-microglobulin protein in A549 and NCI-H1299 lung cancer cells was measured by Western blot assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Error bars indicate the SD.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Asterisks indicate p < .05, significantly different compared with the control; Hashtags indicate p < .05, significantly different compared with the group treated with 6.25 μg/ml Endostar; ampersands indicate p < .05, significantly different compared with the group treated with 12.5 μg/ml Endostar; one-way analysis of variance followed by LSD test.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The loss of MHC class I expression was probably caused by high level of HIF-1.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After a treatment of A549 lung cancer cells with Endostar for 24 hours, the relative levels of MHC-I α heavy chain mRNA demonstrated a slight enhancement without statistical significance (p = .840) in the groups treated with 6.25 μg/ml, 12.5 μg/ml, and 25 μg/ml Endostar compared with the control group (Figure 5 in detail).

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

The relative levels of MHC-I protein α heavy chain in the A549 cells in the groups treated with 6.25 μg/ml, 12.5 μg/ml, and 25 μg/ml Endostar were statistically enhanced (p = .048, p = .007, p = .002, respectively) compared with the control group, and in the NCI-H1299 cells in the groups treated with 6.25 μg/ml, 12.5 μg/ml and 25 μg/ml Endostar were statistically enhanced as well (p = .008, p = .007, p = .005, respectively) compared with the control group (Figure 6 in detail).

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Figure 5.Effects of Endostar on the mRNA of MHC class I α-heavy chain in A549 lung cancer cells measured by RT-qPCR assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

After treatment with Endostar for 24 h, the mRNA of MHC class I α-heavy chain in A549 lung cancer cells was measured by RT-qPCR assay.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Error bars indicate the SD.

PMC12101583

Inhibitory effect of Endostar on HIF-1 with upregulation of MHC-I in lung cancer cells.

Slight enhancement without statistical significance were found among the groups with Endostar treatment (p > .05); one-way analysis of variance.