南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (01): 125-.

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抑制miR-23a表达增强卵巢癌顺铂敏感性的分子机制

金爱红,周霞平,周凤珍   

  • 出版日期:2015-01-20 发布日期:2015-01-20

Inhibition of microRNA-23a increases cisplatin sensitivity of ovarian cancer cells: the
possible molecular mechanisms

  • Online:2015-01-20 Published:2015-01-20

摘要: 目的分析抑制miR-23a表达后,耐药卵巢癌A2780细胞对化疗药物顺铂的敏感性变化及其分子机制。方法选择顺铂耐
药卵巢癌A2780细胞株为研究样本,并分为两组,其中对照组仅加入顺铂,实验组加入顺铂及antagomir-23a。应用MTT法检测
antagomir-23a抑制miR-23a并经顺铂处理后细胞增殖抑制率;应用流式细胞仪分析细胞周期分布情况;应用Hoechst33258染色
分析细胞凋亡形态学变化;应用Western blot法分析耐药糖蛋白P-gp的表达变化。结果抑制miR-23a并经顺铂处理后,细胞增
殖抑制率显著上升(P<0.01),顺铂中效浓度(IC50)为17.89 μmol/L,比对照组IC50 110.18 μmol/L降低了83.76%(P<0.01),细胞被
阻滞于G0/G1期且凋亡率增加(P<0.01);Hoechst33258染色见细胞核浓缩、染色增强。Western blot检测提示细胞P-gp蛋白表
达随着顺铂浓度加大而逐渐减低(P<0.01)。结论抑制耐药卵巢癌A2780细胞内miR-23a表达后,细胞对顺铂的敏感性显著增
加,这可能miR-23a靶基因负性调控因素得以缓解,并由此引发P-gp蛋白表达受抑有关。

Abstract: Objective To investigate the changes in cisplatin sensitivity of resistant ovarian cancer A2780 cells after inhibition of
miR-23a expression and explore the molecular mechanisms. Methods The drug-resistant ovarian cancer A2780 cells were
exposed to cisplatin alone or in combination with antagomir-23a. The cell inhibition rates after the treatments were detected
using MTT assay, cell cycle changes assessed with flow cytometry, and apoptotic cells observed using Hoechst33258 staining.
The changes in glycoprotein P-gp expression in the cells were detected using Western blotting. Results Inhibition of miR-23a
combined with cisplatin treatment significantly increased the cell inhibition rate (P<0.01) and lowered the IC50 of cisplatin by
83.76% from 110.18 μmol/L in the control group to 17.89 μmol/L (P<0.01). The combined treatments also caused cell cycle arrest
in G0/G1 phase, increased the cell apoptosis rate (P<0.01) and the number of cells stained with Hoechst33258; the cellular
expression of P-gp protein was significantly reduced as the cisplatin doses increased (P<0.01). Conclusion Inhibition of
miR-23a expression increases the sensitivity of A2780 cells to cisplatin possibly by inhibiting the negative regulation by
miR-23a target genes that causes inhibition of P-gp protein expression.