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PMC12172538
Methyltransferase gene-mediated antiproliferative effect of 1,25(OH)2D3 on neuroblastoma cells
In another study, methyltransferase DNMT3B was identified as a factor that stimulates the proliferation of colorectal cancer cells.
PMC12172538
Methyltransferase gene-mediated antiproliferative effect of 1,25(OH)2D3 on neuroblastoma cells
A study has demonstrated that DNMT3A interacts directly with p53, which results in the suppression of p53-mediated transactivation of the p21 gene.
PMC12172538
Methyltransferase gene-mediated antiproliferative effect of 1,25(OH)2D3 on neuroblastoma cells
In this study, it was observed that 1,25(OH)2D3 altered the expression levels of DNMT3A and DNMT3B, but did not cause any change in PTEN expression level in SH-SY5Y cells.
PMC12172538
Methyltransferase gene-mediated antiproliferative effect of 1,25(OH)2D3 on neuroblastoma cells
These results suggest that the methyltransferase effect of DNMT3A and DNMT3B may be effective on other TSGs and different signalling pathways associated with these genes, but not on PTEN.
PMC12172538
Methyltransferase gene-mediated antiproliferative effect of 1,25(OH)2D3 on neuroblastoma cells
Within the limits of this study, the fact that 1,25(OH)2D3 showed antiproliferative, antimigratory activity on neuroblastoma SH-SY5Y cells and decreased the expression levels of genes involved in methylation may be evidence of its anticancer effect.
PMC12172538
Methyltransferase gene-mediated antiproliferative effect of 1,25(OH)2D3 on neuroblastoma cells
In this respect, further and more detailed investigations, both in vitro and in vivo, should be performed to strengthen the evidence of the anticancer effect of 1,25(OH)2D3 in human neuroblastoma.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1 (pleckstrin homology-like domain, family A, member 1) is a multifunctional protein involved in pathophysiological conditions such as cancer.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Our previous research revealed changes in the proteome of PHLDA1-silenced human neuroblastoma IMR-32 cells.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
One of the proteins that was increased in PHLDA1-silenced cells was the efflux pump ABCB1 (ATP binding cassette subfamily B member 1).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
In this study, ABCB1 efflux pump upregulation in PHLDA1-silenced IMR-32 cells led to increased resistance of the cells to xenobiotic agents.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Experiments revealed that xenograft tumors grown in immunocompromised mice from PHLDA1-silenced cells presented multiple changes in comparison with tumors grown from control cells.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Tumors grown from PHLDA1-silenced cells were larger and contained fewer apoptotic cells than control tumors.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The blood vessels within the tumors grown from PHLDA1-silenced cells displayed increased extravasation and the extracellular matrix of the tumors presented increased collagen content in comparison to that of the control tumors.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The results showed that silencing PHLDA1 in vivo promoted human neuroblastoma cell survival, decreased the apoptotic potential of the cells, disrupted angiogenesis in developing tumors, and altered collagen network formation.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1 (pleckstrin homology-like domain, family A, member 1) is a protein that plays a role in many cellular functions, and owing to its ambiguous involvement in various types of cancers, this protein is a frequent subject of research.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1 is involved in apoptosis and is a confirmed target of the P53 protein.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1 participates in the apoptotic process by inhibiting the PI3K/Akt pathway, leading to caspase activation and the induction of cell death in many human cancer cell lines.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1 expression may be reduced in certain types of cancer, potentially contributing to uncontrolled cancer cell growth.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1 might act as a tumor suppressor and is a negative regulator of aurora A kinase in breast cancer, and downregulation of PHLDA1 mRNA expression is associated with poor prognosis in that cancer.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Moreover, the pleckstrin homology domain is known for binding membrane phospholipids, which suggests that PHLDA1 may be involved in cellular signaling.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1 attracted our interest in previous research.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Its expression was upregulated in IMR-32 human neuroblastoma (NB) cells after anti-GD2 14G2a antibody treatment.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Another study revealed that IMR-32 cells in which the PHLDA1 gene was silenced presented increased cellular ATP concentrations and increased mitochondrial potential and were less prone to apoptosis than control cells were.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Moreover, the level of a marker of poor prognosis in neuroblastoma, TRKB (tropomyosin receptor kinase B), was increased in PHLDA1-silenced cells.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
We showed that silencing of PHLDA1 gene led to a significant increase in the expression of AURKA (encoding Aurora A kinase), increased phosphorylation of p-Aurora A at Thr288 and Akt at Thr308, and lower levels of cleaved PARP and caspase-3 as well as lower activity of apoptosis-executing caspase 3 and − 7.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Thus, the overall results suggested a role of PHLDA1 as a pro-apoptotic protein.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
In addition, a significant decrease in the mRNA level of the P21 protein (a product of the CDKN1A gene, which encodes a cell cycle inhibitor), was noted after PHLDA1 silencing.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
By flow cytometry analysis of propidium iodide stained cells, we measured slightly higher percentage of cells in G0/G1 phase for the PHLDA1-silenced clones with down-regulation of PHLDA1 (ca.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
70%) - as compared to WT and Mock1 cells (ca.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
65%) and marginally lower number of cells in S and G2/M phases, as compared to WT and Mock1 cells.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Our most recent study on the IMR-32 human neuroblastoma cell line revealed noticeable changes in the global proteome and phosphoproteome after PHLDA1 silencing, which led to the upregulation of proteins associated with mitochondria.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Among the proteins identified in PHLDA1-silenced cells, ABCB1 was highly upregulated according to mass spectrometry analysis.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
ABCB1 (other names: MDR1, P-GP1) is an efflux pump that exports xenobiotics and other harmful substances from cells.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
It is strongly involved in the multidrug resistance of cancer cells (MDR).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
In human neuroblastoma, multidrug resistance induced by the activity of various ABC transporters often occurs in the tumors of high-risk patients.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
ABCB1 and several other ABC transporters are regulated directly by the MYCN oncogene, which is a major oncogenic driver in neuroblastoma.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
In this study, the inverse correlation between PHLDA1 and ABCB1 found in our previous studies via mass spectrometry was confirmed via RT-qPCR and western blotting.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The increased level of ABCB1 after PHLDA1 silencing led to the induction of xenobiotic resistance in IMR-32 and CHP-134 cells.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Our in vivo experiments revealed that xenograft tumors derived from PHLDA1-silenced cells were larger and more densely packed than control tumor cells, hemorrhagic, and their collagen network was more abundant than that of tumors grown from control cells.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
These findings point to a role for PHLDA1 in modulating neuroblastoma tumor growth dynamics, ECM remodeling and chemoresistance development.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Our previous study involving global mass spectrometry analysis revealed that the ABCB1 protein, encoding an efflux pump, was found as one of the characteristic significantly enhanced proteome compound in PHLDA1-silenced IMR-32 cells (shP).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
To investigate this finding further, RT‒qPCR and western blot analyses were performed (Fig. 1; Supplementary Fig. S1-S3 online).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The level of the ABCB1 transcript was more than 15-fold greater in PHLDA1-silenced cells (shP) than in control cells (shC) (Fig. 1a).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The relative PHLDA1 gene expression levels are presented in Fig. 1b.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
As previously reported, PHLDA1-silenced cells are characterized by a significantly decreased level of the PHLDA1 transcript.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The decreased level of PHLDA1 protein is shown in Fig. 1c (as detected with the anti-PHLDA1 antibody Sc-23866 from Santa Cruz Biotechnology).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The nature of the upper band was verified in a separate experiment with an additional anti-PHLDA1 antibody (ab133654 from Abcam, Supplementary Fig. S4 online).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Therefore, the upper band shown was labeled as an unspecific band.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Moreover, the level of the ABCB1 protein was easily detected in shP cells, although the protein remained undetectable in control shC and WT cells (Fig. 1c).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Fig. 1Gene expression analysis of PHLDA1-silenced IMR-32 cells (shP), control cells (shC) and wild-type cells (WT) revealed significant changes in ABCB1 (a) and PHLDA1 (b) transcript levels.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The ABCB1 mRNA content in shC was set as 1.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The PHLDA1 mRNA content in the WT was set as 1.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The data are shown as the means (± SEM) of 3 (a) and 4 (b) independent experiments.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The samples were run in triplicate.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Statistical significance was calculated with Student’s t test for independent samples (*p < 0.05, exact p = 0.023) (a) and with Kruskal‒Wallis ANOVA followed by Dunn’s test (*p < 0.05, exact p = 0.042) (b).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Western blot analysis of ABCB1 and PHLDA1 in shP, shC and WT IMR-32 cells (c).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The HepG2 cell line was used as a positive control for the ABCB1 protein.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
α-Tubulin was used as a reference.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Representative immunoblots of 3 independent experiments are shown (*- unspecific band).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Gene expression analysis of PHLDA1-silenced IMR-32 cells (shP), control cells (shC) and wild-type cells (WT) revealed significant changes in ABCB1 (a) and PHLDA1 (b) transcript levels.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The ABCB1 mRNA content in shC was set as 1.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The PHLDA1 mRNA content in the WT was set as 1.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The data are shown as the means (± SEM) of 3 (a) and 4 (b) independent experiments.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The samples were run in triplicate.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Statistical significance was calculated with Student’s t test for independent samples (*p < 0.05, exact p = 0.023) (a) and with Kruskal‒Wallis ANOVA followed by Dunn’s test (*p < 0.05, exact p = 0.042) (b).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Western blot analysis of ABCB1 and PHLDA1 in shP, shC and WT IMR-32 cells (c).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The HepG2 cell line was used as a positive control for the ABCB1 protein.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
α-Tubulin was used as a reference.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Representative immunoblots of 3 independent experiments are shown (*- unspecific band).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The level of ABCB1 following PHLDA1 silencing was assessed in another human neuroblastoma cell line, CHP-134.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1 and ABCB1 levels were measured in PHLDA1-silenced (the S6 and S17 clones), control (the Mock3 clone) and WT cells via western blotting.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The upregulation of ABCB1 following the silencing of PHLDA1 was also confirmed in CHP-134 cells.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The levels of ABCB1 in the S6 and S17 PHLDA1-silenced clones were greater than those in the Mock3 and WT clones after 48 h of culture in vitro (Supplementary Fig. S5 online).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The activity of the ABCB1 protein as an efflux pump was examined via two functional assays: a rhodamine 123 accumulation test and a doxorubicin cytotoxicity test.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Rhodamine 123 is a fluorescent dye that is known to be one of the chemicals actively exported by the ABCB1 protein.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
To test ABCB1 protein activity in our model, shP, shC and WT cells were treated with 1 µM rhodamine 123 solution for 4 h, and the fluorescence was measured.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
shC and WT cells displayed high fluorescence intensity because rhodamine 123 accumulated within the cells.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
In contrast, shP cells were nonfluorescent, i.e., they did not accumulate rhodamine 123.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
This means that rhodamine 123 was actively and efficiently pumped out of the shP cells (Fig. 2a; Supplementary Fig. S6 online).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Fig. 2ABCB1 activity in IMR-32 cells with PHLDA1 silencing.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
ABCB1 activity was verified via a rhodamine 123 accumulation assay (a) and visualized via fluorescence microscopy.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Images of PHLDA1-silenced (shP), control (shC), and WT IMR-32 cells are shown in fluorescence mode, and visible light is presented in the upper and lower panels, respectively.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Scale bars – 100 μm.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Fluorescence microphotographs were taken during all three experiments.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Brightfield microphotographs were taken during one experiment (Supplementary Fig. S6 online).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
ABCB1 functional test after doxorubicin treatment (b).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
PHLDA1-silenced (shP) and control (shC) IMR-32 cells were treated with 30 nM doxorubicin (dox) or water (Ø) for 48 h, protein lysates were subsequently obtained, and western blot analysis was performed in three independent experiments.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Representative immunoblots are shown (*- unspecific band).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
An ATP luminescence test was performed to confirm the cytotoxic effect of doxorubicin (c).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The number of control cells treated with water was set to 1.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The samples were run in triplicate.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
The data are shown as the means (± SEM) of three independent experiments.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Statistical significance was assessed by one-way ANOVA, followed by post hoc Tukey’s test (*p < 0.05, exact p = 0.022 for shP water vs. shC water; ***p < 0.001, exact p = 0.00043 for shC water vs. shC doxorubicin; p = 0.000024 for shP doxorubicin vs. shC doxorubicin).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
ABCB1 activity in IMR-32 cells with PHLDA1 silencing.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
ABCB1 activity was verified via a rhodamine 123 accumulation assay (a) and visualized via fluorescence microscopy.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Images of PHLDA1-silenced (shP), control (shC), and WT IMR-32 cells are shown in fluorescence mode, and visible light is presented in the upper and lower panels, respectively.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Scale bars – 100 μm.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Fluorescence microphotographs were taken during all three experiments.
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
Brightfield microphotographs were taken during one experiment (Supplementary Fig. S6 online).
PMC12738729
PHLDA1 silencing in IMR-32 human neuroblastoma cells results in ABCB1 overexpression, augments chemoresistance and leads to increased growth of tumors
ABCB1 functional test after doxorubicin treatment (b).