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miR-148a靶向STAT3对宫颈癌HeLa细胞顺铂化疗敏感度的增强作用

Enhanced Effect of miR-148a Targeting STAT3 on Chemosensitivity of Cervical Cancer HeLa Cells to Cisplatin

  • 摘要:
    目的 探究miR-148a对宫颈癌HeLa细胞顺铂化疗敏感度的影响及相关机制。
    方法 体外培养宫颈癌HeLa细胞,设置顺铂浓度梯度检测IC20值;设置对照组、mimic对照组、miR-148a mimic组、inhibitor对照组和miR-148a inhibitor组,转染后使用qRT-PCR法检测miR-148a和STAT3 mRNA表达;4 μmol/L顺铂处理后,MTT法检测细胞增殖,流式细胞仪检测细胞凋亡及细胞周期分布,Western blot法检测p-STAT3/STAT3、CyclinD1、Bcl-2、Bax、Cleaved caspase-3蛋白表达。
    结果 HeLa细胞的顺铂IC20约为4 μmol/L。与mimic对照组相比,miR-148a mimic组HeLa细胞中miR-148a水平显著升高,STAT3 mRNA水平显著降低(P < 0.05);与inhibitor对照组相比,miR-148a inhibitor组HeLa细胞中miR-148a水平显著降低,STAT3 mRNA水平显著升高(P < 0.05)。顺铂处理后,与mimic对照组相比,miR-148a mimic组HeLa细胞凋亡率、G0/G1期细胞比例、Bax、Cleaved caspase-3蛋白水平显著升高,OD值、S期和G2/M期细胞比例及p-STAT3/STAT3、Cyclin D1、Bcl-2蛋白水平显著降低(P < 0.05);与inhibitor对照组相比,miR-148a inhibitor组HeLa细胞上述指标均朝相反的趋势显著变化(P < 0.05)。
    结论 miR-148a可通过靶向抑制STAT3增强宫颈癌HeLa细胞的顺铂化疗敏感度。

     

    Abstract:
    Objective To investigate the effect of miR-148a on the chemosensitivity of cervical cancer HeLa cells to cisplatin and its related mechanism.
    Methods Cervical cancer HeLa cells were cultured in vitro and the concentration gradient of cisplatin was set to detect IC20 value. Control group, mimic control group, miR-148a mimic group, inhibitor control group and miR-148a inhibitor group were set up. qRT-PCR was used to detect the expression of miR-148a and STAT3 mRNA after transfection. After 4 μmol/L cisplatin treatment, the proliferation of HeLa cells was detected by MTT assay; the apoptosis and cell cycle distribution were detected by flow cytometry; Western blot was used to detect the protein expression of p-STAT3/STAT3, CyclinD1, Bcl-2, Bax and Cleaved caspase-3.
    Results The IC20 of cisplatin on HeLa cells was about 4 μmol/L. Compared with the mimic control group, the level of miR-148a in the miR-148a mimic group was significantly increased, and the level of STAT3 mRNA was significantly decreased (P < 0.05). Compared with the inhibitor control group, the level of miR-148a in HeLa cells in miR-148a inhibitor group was significantly decreased, and the level of STAT3 mRNA was significantly increased (P < 0.05). On the basis of cisplatin treatment, compared with the mimic control group, the apoptosis rate, G0/G1 phase cell ratio, the protein levels of Bax and Cleaved caspase-3 were significantly increased in miR-148a mimic group, while OD value, the proportions of cells in S and G2/M phase, the protein levels of p-STAT3/STAT3, CyclinD1, Bcl-2 were significantly decreased (P < 0.05); compared with the inhibitor control group, the above indicators of HeLa cells in miR-148a inhibitor group changed significantly in the opposite direction (P < 0.05).
    Conclusion MiR-148a could enhance the chemosensitivity of cervical cancer HeLa cells to cisplatin by targetedly inhibiting STAT3.

     

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