PMCID string | Title string | Sentences string |
|---|---|---|
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | Seven-week-old male BALB/c-nude mice were purchased from Orient Bio (Gyeonggi, South Korea). |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | A total of 1 × 10 GIST430 or GIST430-V654A cells were mixed with an equal volume of Matrigel (BD Biosciences), and each mouse was subcutaneously injected with 0.1 ml of the cell-Matrigel mixture into the flank. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | Tumor volume was measured using a digital Vernier caliper (Mitutoyo; Kanagawa, Japan) three times a week after implantation and was calculated as 1/2 × length × width . |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | When the tumor volume had reached 200–300 mm about 3 weeks, the mice were randomized into five groups (n = 4–6/group) according to cell line and administered an intraperitoneal injection of 2% DMSO in PBS (vehicle), 50 mg kg imatinib, 30 mg kg PF429242, 1 mg kg bortezomib, 30 mg kg PF429242, and 1 mg kg bortezomib. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | Eight injections were performed at three-day intervals, for a total of eight injections. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | Body weight and tumor volume were measured three times a week after drug treatment. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | At the end of the experiments, the animals were sacrificed, and the tumors were isolated, weighed, and stored at −80 °C until further analyses. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | Tumors were paraffin-embedded and 4 μm sections were used for immunohistochemistry. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | The sections were deparaffinized and treated with BLOXALL endogenous blocking solution (Vector Laboratories, CA, USA) to inhibit endogenous peroxidase activity. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | The epitope retrieval was conducted with Retrieve-All Antigen 1 (pH 8), followed by blocking with PBS supplemented with 2% normal horse serum and 3% BSA. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | Sections were incubated with individual primary antibodies at 4 °C overnight and with the appropriate secondary antibody at room temperature for 1 h. Peroxidase activity was revealed using 3,3-diaminobenzidine (Vector Laboratories), and sections were counterstained with hematoxylin (Vector Laboratories). |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | After dehydrogenation with alcohol and xylene, sections were viewed and photographed with an Olympus BX51 microscope (Olympus, Tokyo, Japan) and Mosaic2.1 software. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | The following antibodies were used: Ki-67 (1:100, Abcam, Cambridge, UK), cleaved caspase-3 (1:100, Cell Signaling Technology, MA, USA), and KIT (1:200, Abcam). |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | Ki-67, cleaved caspase-3, and KIT. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | Positive staining in xenografted tumors were quantified using ImageJ software (NIH). |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | Three HPF (high-power fields, at ×400 magnification) were used for each tumor . |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | All data were analyzed using GraphPad Prism 7 (GraphPad Software Inc., La Jolla, CA, USA) and expressed as mean ± SEM. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | A comparison of the means among groups was performed using Mann-Whitney U test for body weight, tumor weight, and tumor volume data, or one-way analysis of variance (ANOVA) followed by Dunnett’s post-hoc test for IHC data. |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | Fisher’s exact test, Student’s t test, and Kaplan–Meier survival analysis were used to analyze the relationship between nuclear ATF6 expression and clinicopathological parameters (SPSS software, SPSS, Inc., Chicago, IL, USA). |
PMC10589262 | Identification of novel pathogenic roles of BLZF1/ATF6 in tumorigenesis of gastrointestinal stromal tumor showing Golgi-localized mutant KIT | Statistical significance was set at P < 0.05. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | •Knockdown of circAPP inhibits GIST proliferation and the Warburg effect.•miR-6838-5p induces GIST proliferation and the Warburg effect.•CircAPP activates GIST proliferation and the Warburg effect by regulating the miR-6838-5p/CDV3 axis. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Knockdown of circAPP inhibits GIST proliferation and the Warburg effect. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | miR-6838-5p induces GIST proliferation and the Warburg effect. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | CircAPP activates GIST proliferation and the Warburg effect by regulating the miR-6838-5p/CDV3 axis. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Gastrointestinal Stromal Tumor (GIST) occurs mainly in the stomach and small intestine but also in the colon and rectum, esophagus, and other organs. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Mutations that activate Tyrosine Kinase (KIT) or platelet-derived growth factor receptor α are reported to be the main molecular mechanism of GIST. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Currently, KIT inhibitor (imatinib) is the first-line chemotherapeutic drug for GIST treatment, but most GIST patients develop imatinib resistance. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Although key aspects of the pathogenesis of GIST have been elucidated, the mechanism of oncogene overexpression and gene expression regulators in GIST remains unclear. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Metabolic reprogramming is the earliest known cancer hallmark, which achieves tumor cell proliferation and survival by altering bioenergetic and biosynthetic pathways, and tumor cells exhibit a high rate of glycolysis, also known as the Warburg effect. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Therefore, exploring the regulatory mechanism of the Warburg effect is likely to develop new strategies for GIST treatment. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | CircRNAs are endogenous non-coding RNAs that differ from traditional linear RNAs in that they have covalently closed structures. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | CircRNA/microRNAs (miRNAs) interaction can regulate gene expression and participate in various physiological and pathological processes. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Furthermore, due to the high stability and conservation of circRNAs and their high abundance in body fluids, circRNAs have been considered potential biomarkers for GIST. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | CircRNA Amyloid Precursor Protein (circAPP) is an abnormally highly expressed circRNA in GIST. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | In the preliminary experiments, it was also determined that circAPP was abnormally highly expressed in GIST. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Therefore, this study focused on exploring the function and potential downstream molecular mechanisms of circAPP in GIST. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | CircRNAs expressed in the cytoplasm contain miRNA binding sites to act as miRNA sponges, preventing miRNAs from interacting with mRNAs in the 3′Untranslated Region (UTR), thereby modifying miRNA downstream mRNAs, thereby affecting a series of cellular activities. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | miRNAs are capable of modifying post-transcriptional gene expression by recognizing and binding to the 3′-UTR of target gene mRNAs. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Dysregulated miRNAs are key to many cancers including GIST, where they act as tumor suppressors or oncogenes. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | For example, miR-4510 promotes GIST cell invasion and metastasis whereas miR-494 suppresses GIST development. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | miR-6838-5p has been confirmed to be involved in osteosarcoma, gastric cancer, and renal cell carcinoma. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | However, no studies have demonstrated its actions in GIST. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Targeting to unearth the mechanism of circAPP in GIST, various studies were included, ultimately discovering the mediatory impacts of circAPP on proliferation, migration, and Warburg effect through miR-6838-5p targeting CDV3 in GIST, and hopefully replenishing the options for molecule-targeted therapy in GIST. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Between April 2013 and July 2015, 52 pairs of GIST tissue samples and adjacent normal tissue (≥ 5 cm from cancer tissue) were obtained from the Affiliated Hospital of Inner Mongolia Minzu University. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Inclusion criteria: 1) All the included subjects met the diagnostic criteria of GIST according to the criteria of “Chinese Expert Consensus on Diagnosis and Treatment of Gastrointestinal Mesenchymal Tumor (2017 Edition)” and “NIH2008 Modified Chinese Consensus 2017 Revised”; 2) Patients with GIST observed by abdominal CT or ultrasound endoscopy with tumor located in the stomach, and those who were diagnosed by pathology; 3) Clinical data of all the study subjects must be accurate and complete; 4) Patients not undergoing any radiotherapy or chemotherapy; 5) Patients with no history of other concomitant malignant tumors. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | With written informed consent, tissue sampling was done with the approval of the Ethics Committee of the Affiliated Hospital of Inner Mongolia Minzu University (n° 201211Y62). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | After diagnosis by a pathologist, tissue specimens were frozen in liquid nitrogen and then stored at -80 °C to determine gene expression. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | All methods in the study were carried out in accordance with the ARRIVE guidelines. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | The human gastric mucosal epithelial cell line GES-1 and the human GIST cell lines GIST-882 and GIST-T1 (Cosmo, Tokyo, Japan) were serially passaged for no more than 6 months. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Cells were cultured in DMEM (Gibco; Thermo Fisher Scientific) in a humidified incubator at 37 °C with 5 % CO2. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | 10 % heat-inactivated fetal bovine serum (Gibco; Thermo Fisher Scientific), 100 U/mL penicillin, and 100 mg/mL streptomycin were supplementary to the culture medium. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | The cell lines were all validated by STR profiles and tested negative for mycoplasma contamination. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Under the guidelines of TRIzol (Invitrogen), total RNA was extracted. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | For reverse transcription, the PrimeScript RT kit (Takara, Shiga, Japan) was employed. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Taking cDNA as a template, RNA quantification was done using specific primers (Table 1) and TB Green qPCR Master Mix (Takara) on a 480 II instrument (Roche Diagnostics, Basel, Switzerland). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Expression levels of circRNA, mRNA, and miRNA were calculated by the 2 method and standardized to GAPDH or U6.Table 1Primers. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Table 1PrimerscircAPPForward: 5′-TGACTGAAGGGAAGTGTGCC-3′Reverse: 5′-CAAACTCACCAATGGCGCTG-3′miR-6838-5pForward: 5′-GCAAGCAGCAGTGGCAAG-3′Reverse: 5′-GCAGGGTCCGAGGTATTC-3′GAPDHForward: 5′- CACCCACTCCTCCACCTTTG-3′Reverse: 5′-CCACCACCCTGTTGCTGTAG-3′U6Forward: 5′-CTCGCTTCGGCAGCACA-3′Reverse: 5′-AACGCTTCACGAATTTGCGT-3′Note: circAPP, circular RNA APP; miR-6838-5p, microRNA-6838-5p; GAPDH, Glyceraldehyde-3-Phosphate Dehydrogenase. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Primers. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Note: circAPP, circular RNA APP; miR-6838-5p, microRNA-6838-5p; GAPDH, Glyceraldehyde-3-Phosphate Dehydrogenase. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | For RNase R treatment, total RNA (2 μg) was cultured with 3 U/μg RNase R (Geneseed Biotech, Guangzhou, China) at 37°C for 10 min. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Then, RNA purification was done with the RNeasy MinElute Cleanup kit (Qiagen, Hilden, Germany). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | For actinomycin D treatment, RNA was treated with 2 mg/mL actinomycin D (Sigma-Aldrich). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Dimethyl sulfoxide was taken as a control. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Finally, RNA expression levels were quantitatively analyzed. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Subcellular localization of circAPP in GIST-882 cells was analyzed. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Nuclear and cytoplasmic fractions were isolated using the PARIS™ kit (Invitrogen), with U6 and 18S rRNA as positive references, respectively. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Small interfering RNA targeting circAPP and CDV3, small interfering RNA negative control (si-circAPP: 5’-ATGGATGTTTGCGAAACTCATCT-3’, si-CDV3, si-NC), overexpression plasmid targeting circAPP and CDV3 and negative control (pcDNA 3.1-circAPP/CDV3, pcDNA 3.1) were supplied by RiboBio (Guangzhou, China), whereas miR-6838-5p mimic (5’-AAGCAGCAGTGGCAAGACTCCT-3’)/inhibitor and mimic/inhibitor NC were by Genepharma (Shanghai, China). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Lipofectamine 2000 (Invitrogen) served for the transfection of cells seeded at 2×10/well on 6-well plates. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Briefly, when cells were 70-80 % confluent, oligonucleotides or plasmids were diluted in Opti-MEM® Medium. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Lipofectamine 2000 was then diluted with Opti-MEM® Medium. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Lipofectamine 2000 was mixed with the diluted oligonucleotide or plasmid and added to each well at 100 μL/well. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | After 48h, the transfection efficiency was determined by RT-qPCR or western blot. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Cell samples from log phase cultures were transferred to 6-well plates at 500 cells/well and cultured for 14 days. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Afterward, samples were fixed in 4 % paraformaldehyde and counted after 0.1 % crystal violet staining. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | GIST-882 and GES-1 cells were incubated with 30 μM EdU (KeyGen Biotech, Jiangsu, China) for 2h and treated with DAPI (Sigma-Aldrich) to stain the nuclear. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Fluorescent images were taken to assess the EdU-positive rate. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Apoptotic cells were assessed with an annexin V-FITC/PI kit (Vazyme, Nanjing, China). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Cells were resuspended and stained with Annexin V-FITC and PI solutions (5 μL each solution) in the dark. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Apoptotic cells were detected on a FACSCanto II flow cytometer (BD Biosciences) to calculate the apoptosis rate. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Glucose consumption, lactate level, and ATP were determined by glucose assay kits (Sigma), lactate colorimetric/fluorometric kits (BioVision, CA, USA), and ATP assay kits (Thermo Fisher Scientific), respectively. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | ATP/ADP was measured using the ApoSENSOR ADP/ATP Ratio Assay Kit (K255-200, BioVision, CA, USA) and luminescence was measured by spectroscopy (Molecular Devices, CA USA). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Cells (10 cells) were seeded onto luminometer plates, incubated with nucleotide release buffer, and added with 1 µL of ATP monitoring enzyme. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Data were recorded at 1 min (Data A) and 10 min (Data B). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | ADP convertase was then added, and values were read (Data C). . |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | NAD/NADH was tested using the EnzyChrom™ NAD/NADH Ratio Assay Kit (E2ND-100, Bioassay Systems, CA, USA). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | A total of 10 cells were homogenized and cultured with 100 mL NAD extraction buffer for NAD assay or 100 mL NADH extraction buffer for NADH assay. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Next, 20 µL of assay buffer was added to and 100 mL of back-extraction buffer was then supplemented. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | The mixture was then centrifuged, and the supernatant was transferred to the working reagent. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | OD565 nm was read at 0 and 15 min. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Total protein from cells and tissues was isolated using Lysis Buffer (Thermo Fisher Scientific) and analyzed for concentration using the BCA Assay Kit (Bio-Rad). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Proteins (30 μg) were separated by 10 % SDS-PAGE and then transferred to PVFD membranes (Millipore, MA, USA). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Membranes were blocked with 5 % skim milk, mixed with primary antibodies GAPDH (60004-1-Ig, Proteintech), CDV3 (LS-C205853, LSBio), HK2 (22029-1-AP, Proteintech), PKM2 (15822-1-AP, Proteintech), followed by the secondary antibody (Sigma-Aldrich). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | To visualize protein bands, a Western blot substrate (Thermo Fisher Scientific) was added, and Image Lab analysis software (Bio-Rad) was employed for reading protein band intensity. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | CircAPP and CDV3-3′UTR wild-type sequences (WT-circAPP, WT-CDV3) or mutant sequences (MUT-circAPP, MUT-CDV3) containing the miR-6838-5p binding site were cloned into the vector psiCHECK-2. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | The above-mentioned reporter gene vector, along with miR-6838-5p mimic or mimic NC was co-transfected into GIST-882 cells (5×10) in the 24-well plates using Lipofectamine 2000 (Invitrogen). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Cells were lysed after 48h, and luciferase activity was evaluated using Dual-Luciferase Assay kit (Promega). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | The RIP assay was performed using the Imprint® RIP Kit (Sigma-Aldrich). |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | GIST-882 cells (1×10) were lysed with RIP buffers and interacted with antibody-coated magnetic beads at 4°C overnight. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | Anti-IgG was the control for Anti-Ago2. |
PMC11269792 | CircRNA amyloid precursor protein by competitive adsorption of microRNA-6838-5p mediates CDV3 expression to enhance malignant behavior and Warburg effect in Gastrointestinal Stromal Tumor | After RNA isolation from the magnetic beads, analysis of RNA expression was carried out by RT-Qpcr. |
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