人表皮生长因子受体2对卵巢癌细胞凋亡、侵袭的影响及机制研究

Effect of human epidermal growth factor receptor 2 on the apoptosis and invasion of ovarian cancer cell and its mechanism

目的 探讨人表皮生长因子受体2(HER2)对卵巢癌细胞A2780和SKOV3凋亡、侵袭的影响及机制。方法 收集27例卵巢癌患者的27对卵巢癌组织和癌旁组织作为组织样本,选取卵巢癌细胞(A2780细胞和SKOV3细胞)、卵巢上皮细胞HOSEpiC作为实验细胞。采用实时定量聚合酶链反应(qRT-PCR)、蛋白免疫印迹法、免疫组织化学技术检测HER2、E74样ETS转录因子3(ELF3)、音猬因子(SHH)、GLIS家族锌指蛋白1(GLI1)等表达水平;采用细胞计数试剂盒8(CCK-8)法、流式细胞仪和Transwell小室实验检测A2780细胞和SKOV3细胞的活力、凋亡率和侵袭能力。结果 HER2蛋白和HER2 mRNA在卵巢癌组织中的表达均高于癌旁组织,HER2蛋白在A2780细胞、SKOV3细胞中的表达均高于HOSEPiC细胞,差异有统计学意义(P<0.05)。A2780、SKOV3细胞中,与转染si-NC的细胞相比,转染si-HER2的细胞HER2蛋白表达水平降低,且细胞活力降低、细胞凋亡率增加、侵袭细胞数减少,差异有统计学意义(P<0.05)。A2780、SKOV3细胞中,转染si-HER2的细胞ELF3 mRNA表达均低于转染si-NC的细胞,且转染si-HER2+ELF3的细胞ELF3 mRNA表达均高于转染si-HER2+pcDNA的细胞,差异有统计学意义(P<0.05);与转染si-NC的细胞相比,转染si-HER2的细胞活力降低、细胞凋亡率增加、侵袭细胞数减少,差异有统计学意义(P<0.05);与转染si-HER2+pcDNA的细胞相比,转染si-HER2+ELF3的细胞活力增加、细胞凋亡率降低、侵袭细胞数增加,差异有统计学意义(P<0.05)。A2780、SKOV3细胞中,转染si-HER2的细胞SHH、GLI1蛋白表达均低于转染si-NC的细胞,转染si-HER2+ELF3的细胞SHH、GLI1蛋白表达均高于转染si-HER2+pcDNA的细胞,差异有统计学意义(P<0.05)。结论 HER2沉默可通过调控ELF3/SHH通路促进卵巢癌细胞凋亡和抑制卵巢癌细胞侵袭,靶向抑制HER2或可成为治疗卵巢癌的一种有效策略。

Objective To investigate the effect of human epidermal growth factor receptor 2(HER2)on apoptosis and invasion of ovarian cancer A2780 and SKOV3 cells and its mechanism.Methods A total of 27 pairs of ovarian tissues and paracancerous tissues were collected from 27 ovarian cancer patients.Ovarian cancer cells(A2780 cells and SKOV3 cells)and ovarian epithelial cells HOSEpiC were selected as experimental cells.Real-time quantitative polymerase chain reaction(qRT-PCR),Western blot and immunohistochemical techniques were used to detect the expression levels of HER2,E74-like ETS transcription factor 3(ELF3),sonic hedgehog(SHH)and GLIS family zinc finger protein 1(GLI1);cell viability,apoptotic rate and invasive ability of A2780 and SKOV3 cells were analyzed by cell counting kit-8(CCK-8),flow cytometry and Transwell assay,respectively.Results HER2 protein and HER2 mRNA expression were higher in ovarian cancer tissues than in paracancerous tissues,the expression of HER2 protein in A2780 cells and SKOV3 cells was higher than that in HOSEPiC cells(P<0.05).In A2780 and SKOV3 cells,compared with cells transfected with si-NC,the expression level of HER2 protein was decreased in cells transfected with si-HER2,and the cell viability was decreased,the cell apoptosis rate was increased,and the number of invasive cells was decreased(P<0.05).In A2780 and SKOV3 cells,the expression of ELF3 mRNA in cells transfected with si-HER2 was lower than that in cells transfected with si-NC,and the expression of ELF3 mRNA in cells transfected with si-HER2+ELF3 was higher than that in cells transfected with si-HER2+pcDNA(P<0.05).Compared with cells transfected with si-NC,the viability of cells transfected with si-HER2 decreased,the apoptosis rate increased,and the number of invasive cells decreased(P<0.05).Compared with cells transfected with si-HER2+pcDNA,cells transfected with si-HER2+ELF3 had increased viability,decreased apoptosis rate and increased number of invasion cell(P<0.05).In A2780 and SKOV3 cells,the expression of SHH and GLI1 protein in cells transfected with si-HER2 was lower than that in cells transfected with si-NC,and the expression of SHH and GLI1 protein in cells transfected with si-HER2+ELF3 was higher than that in cells transfected with si-HER2+pcDNA(P<0.05).Conclusion HER2 silencing can promote ovarian cancer cell apoptosis and inhibit cell invasion by regulating the ELF3/SHH pathway,which suggests that inhibition of HER2 expression might be an effective strategy for the treatment of ovarian cancer.