ΜiR-202靶向调节EGFR介导的PI3K/AKT信号通路在紫杉醇治疗卵巢癌中的机制研究

Research on Mechanism of Targeted Adjustment of the MiR-202 for EGFR Mediated PI3K/AKT Signaling Pathway in the Treatment of Ovarian Cancer with Paclitaxel

[目的]探讨miR-202靶向调节EGFR介导的PI3K/AKT信号通路在紫杉醇治疗卵巢癌中的作用机制。[方法]以人卵巢癌细胞株A2780细胞作为研究对象,分别转染miR-NC、miR-202mimics及shEGFR。采用MTT和Transwell实验检测各组细胞增殖、迁移及侵袭能力。采用qRTPCR检测细胞中miR-202和EGFR mRNA表达量。采用Western blot检测EGFR、p-PI3K、PI3K、pAKT和AKT蛋白水平。[结果] qRT-PCR结果显示,与正常人卵巢上皮细胞HOSEpiC相比,miR-202在卵巢癌细胞A2780/WT及A2780/PTX中明显低表达(P<0.05);EGFR mRNA水平显著性上调(P<0.05)。转染miR-202 mimics后,卵巢癌细胞中miR-202相对表达量显著性高于miR-NC组和shEGFR组(P<0.05);而miR-202 mimics组和shEGFR组EGFR mRNA含量显著性低于miR-NC组(P<0.05)。过表达miR-202及转染shEGFR后,miR-202 mimics组和shEGFR组细胞增殖能力较miR-NC组均显著性降低(P<0.05);迁移细胞和侵袭数目也随之减少(P<0.05);且对紫杉醇的半数抑制浓度IC50降低(P<0.05)。免疫印迹测定结果显示,miR-202 mimics组和shEGFR组中EGFR蛋白、p-PI3K/PI3K及p-AKT/AKT磷酸化水平较miR-NC组明显下调(P<0.05)。[结论] miR-202能靶向调节EGFR表达,并且影响卵巢癌细胞的增殖、迁移及侵袭过程,其作用机制是通过抑制EGFR下游PI3K/AKT信号通路从而发挥抑癌作用,为临床治疗提供了新方向。

[Objective] To explore the mechanism of targeted adjustment of miR-202 for EGFR mediated PI3 K/AKT signaling pathway in the treatment of ovarian cancer with paclitaxel. [Methods] The human ovarian cancer cell lines-A2780 cells were selected as the research objects,and transfected with miR-NC,miR-202 mimics and sh EGFR. MTT and Transwell experiments were used to determine the cell proliferation,migration and invasion ability in different groups. q RT-PCR was employed to detect the miR-202 level and EGFR m RNA expression. Meanwhile,Western blot was used to test EGFR,pPI3 K,PI3 K,p-AKT and AKT protein levels. [Results] q RT-PCR results showed that miR-202 levels in A2780/WT and A2780/PTX clearly decreased compared with normal ovarian epithelial cells HOSEpi C(P<0.05),and EGFR m RNA levels increased significantly(P<0.05). After transfecting with miR-202 mimics,miR-202 expression in ovarian cancer cells was significantly higher than that in miR-NC group and sh EGFR group(P<0.05). However,EGFR m RNA expression in miR-202 mimics and sh EGFR groups were significantly higher than that in miR-NC group(P<0.05). Due to the overexpression of miR-202 and transfection sh EGFR,the cell proliferation ability of miR-202 mimics and sh EGFR groups cell were significantly lower than that in miR-NC group(P<0.05). Moreover,the IC50 of paclitaxel obviously decreased(P<0.05). There were less migration and invasion cells(P<0.05). Western blot results showed that EGFR protein,p-PI3 K/PI3 K and p-AKT/AKT phosphorylation levels in miR-202 mimics and sh EGFR groups were significantly lower than that in miR-NC group(P<0.05). [Conclusion] miR-202 target adjusts the expression of EGFR to affect cell proliferation,migration and attack process of ovarian cancer,which functions mainly via inhibiting EGFR downstream of PI3 K/AKT signaling pathways and thus plays a role in tumor suppressor,which provides a new direction for clinical treatment.