MicroRNA-21 modulates chemosensitivity of breast cancer cells to doxorubicin by targeting PTEN.

Ovexpression of microRNA-21 (miR-21) is found in various human cancers. Our aim is to investigate the association of miR-21 expression with the sensitivity of breast cancer cells to doxorubicin (ADR). The half maximal inhibitory concentration (IC(50)) value of ADR in resistant MCF-7/ADR or parental MCF-7 cells was determined by MTT assay. TaqMan RT-PCR or Western blot assay was performed to detect the expression of mature miR-21 and tumor suppressor gene (PTEN) protein. MCF-7 or MCF-7/ADR cell line was transfected with miR-21mimic or inhibitor. The IC(50) value of ADR was determined. Flow cytometry and TUNEL assays were performed to analyze apoptosis. The activity of caspase-3 was analyzed. The IC(50) of ADR in MCF-7 and MCF-7/ADR cells was 0.21 ± 0.05 and 16.5 ± 0.08 μmol/L, respectively. We showed that upregulation of miR-21 in MCF-7/ADR cells was concurrent with downregulation of PTEN protein. MiR-21 mimic or inhibitor could obviously affect the sensitivity of breast cancer cells to ADR. Moreover, miR-21 inhibitor could enhance caspase-3-dependent apoptosis in MCF-7/ADR cells. Overexpression of PTEN could mimic the same effects of miR-21 inhibitor in MCF-7/ADR cells and PTEN-siRNA could increase the resistance of MCF-7 cells to ADR. MiR-21 inhibitor could increase PTEN protein expression and the luciferase activity of a PTEN 3' untranslated region-based reporter construct in MCF-7/ADR cells. PTEN-siRNA could partially reverse the increased chemosensitivity of MCF-7/ADR cells induced by miR-21 inhibitor. Dysregulation of miR-21 plays critical roles in the ADR resistance of breast cancer, at least in part via targeting PTEN.

Wang ZX ，Lu BB ，Wang H ，Cheng ZX ，Yin YM ... -  《-》

miR-222 induces Adriamycin resistance in breast cancer through PTEN/Akt/p27(kip1) pathway.

Wang DD ，Yang SJ ，Chen X ，Shen HY ，Luo LJ ，Zhang XH ，Zhong SL ，Zhao JH ，Tang JH ... -  《-》

MiR-222 promotes drug-resistance of breast cancer cells to adriamycin via modulation of PTEN/Akt/FOXO1 pathway.

Acquisition of resistance to adriamycin (ADR) is one of the most important clinical obstacles in the treatment of breast cancer, but the molecular mechanisms underlying sensitivity to ADR remain elusive. In our previous study, through miRNA microarray and experiments, we have emphasized that miR-222 could promote the ADR-resistance in breast cancer cells. The aim of this study was to explore the possible mechanism by which miR-222 affects sensitivity to ADR. Through pathway enrichment analyses for miR-222, we found that PTEN/Akt/FOXO1 signaling pathway may be of importance. RT-qPCR analyses and western blot assays confirmed the relationship between miR-222 expression and target genes. Immunofluorescence further visually displayed the location of FOXO1. When blocking PTEN/Akt/FOXO1 signaling pathway, we demonstrated the effects of miR-222-mediated ADR resistance by MTT and apoptosis assays. RT-qPCR and Western blot results showed that miR-222 expression was negatively correlated with FOXO1 expression. In addition, the subcellular translocation of FOXO1 due to the altered expression of miR-222 was observed from immunofluorescence. Moreover, upregulation of miR-222 expression in MCF-7/S cells is associated with decreased PTEN expression levels and increased phospho-Akt (p-Akt) expression. Conversely in MCF-7/ADR cells, inhibition of miR-222 resulted in increased PTEN expression and decreased p-Akt expression. For further validation, results of the present study also demonstrated that PTEN/Akt/FOXO1 signaling was responsible for the ADR-resistance of breast cancer cells since LY294002, an inhibitor of Akt signaling, partially increased the sensitivity of MCF-7/S cells to ADR. More importantly, we postulated that high expression of miR-222 is closely related to poor overall survival by TCGA database validation. Taken together, these data elucidated that miR-222 mediated ADR-resistance of breast cancer cells partly through regulation of PTEN/Akt/FOXO1 signaling pathway and inhibition of miR-222 may improve the prognosis of breast cancer patients.

Shen H ，Wang D ，Li L ，Yang S ，Chen X ，Zhou S ，Zhong S ，Zhao J ，Tang J ... -  《-》

MicroRNA-34a modulates chemosensitivity of breast cancer cells to adriamycin by targeting Notch1.

MicroRNA-34a (miR-34a) as a tumor suppressor has been reported in many other studies. However, its role in modulating the sensitivity of breast cancer cells to adriamycin (ADR) remains unclear. The aim of this study is to evaluate the role of miR-34a in the sensitivity of breast cancer cells to ADR. The role of miR-34a in breast cancer cells was detected using MTT assay, flow cytometry assay, real-time PCR and Western blot, etc. The association of miR-34a and Notch1 was analyzed by dual-luciferase reporter assay and Notch1-siRNA technology. Real-time PCR assay was performed to test the expression of miR-34a and Notch1 in 38 selective breast cancer tissue samples. Ectopic overexpression of miR-34a could sensitize MCF-7 breast cancer cells to ADR. MiR-34a mimic could inhibit the luciferase activity of the construct containing wild-type 3' UTR of Notch1 in MCF-7/ADR cells. Notch1-siRNA could partially reverse the effect of miR-34a inhibitor in inducing chemoresistance of MCF-7 cells to ADR. Further, there was an inverse association between Notch1 and miR-34a expression in breast cancer. Dysregulation of miR-34a plays critical roles in the acquired ADR resistance of breast cancer, at least in part via targeting Notch1.

Li XJ ，Ji MH ，Zhong SL ，Zha QB ，Xu JJ ，Zhao JH ，Tang JH ... -  《-》

MicroRNA-29a contributes to drug-resistance of breast cancer cells to adriamycin through PTEN/AKT/GSK3β signaling pathway.

Acquisition of resistance to adriamycin (ADR) during the treatment of breast cancer is still a major clinical obstacle. MicroRNAs (miRNAs) are a class of short noncoding RNAs which associate with cancer chemoresistance through regulating gene expression by targeting mRNAs. Our previous microarray found that miR-29a may strongly confer the ADR resistance of breast cancer cells. Here, we aim to explore the possible mechanism by which miR-29a affects sensitivity to ADR. ADR-resistant MCF-7 breast cancer cell subline (MCF-7/ADR) was successfully established in vitro through a stepwise increase of ADR concentrations in the culture based on parental MCF-7 cell lines (MCF-7/S). We used TargetScan (a wide use of target prediction algorithms) in conjunction with pathway enrichment analyses to predict the mRNAs that were most likely to involve in miR-29a-mediated drug resistance in cancers. We confirmed the effects of miR-29a-mediated ADR resistance through MTT and apoptosis assays, and further investigated the activities of two target genes, PTEN and GSK3β, by RT-qPCR analyses and western blot assays. The expression level of miR-29a in MCF-7/ADR cells was remarkablely higher than in MCF-7/S cells. Further MTT and apoptosis assays revealed that transfection of miR-29a inhibitors into MCF-7/ADR cells resulted in prominent reduction of the drug resistance, in contrast, transfection of miR-29a mimics into MCF-7/S cells obviously increased their drug resistance. Through pathway enrichment analyses for miR-29a, we found that PTEN/AKT/GSK3β signaling pathway may be of importance. RT-qPCR and Western blot results showed that downregulation of miR-29a expression in MCF-7/ADR cells increased PTEN expression levels, resulting in decreased phospho-Akt (p-Akt) and phospho-GSK3β (p-GSK3β) expression. Conversely, upregulation of miR-29a expression in MCF-7/S cells is associated with decreasing PTEN expression and increasing p-Akt and p-GSK3β expression. PTEN and GSK3β are targeted by miR-29a, and miR-29a may contribute to ADR resistance through inhibition of the PTEN/AKT/GSK3β pathway in breast cancer cells. Thus, miR-29a may be a potential target for the patients who acquired ADR-resistance during the treatment of breast cancer.

Shen H ，Li L ，Yang S ，Wang D ，Zhong S ，Zhao J ，Tang J ... -  《-》