DUSP6沉默对人子宫内膜癌Ishikawa细胞凋亡和迁移的影响及机制研究

Effect of DUSP6 silencing on apoptosis and migration of human endometrial cancer Ishikawa cells and its mechanism

目的:探讨双特异性磷酸酶6(DUSP6)沉默对人子宫内膜癌Ishikawa细胞凋亡和迁移的影响及机制。方法:Ishikawa细胞随机分为空白对照组(转染脂质体试剂)、过表达组(转染DUSP6)、过表达NC组(转染DUSP6-NC)、沉默组(转染siDUSP6)、沉默NC组(转染siDUSP6-NC)。RT-qPCR检测细胞中DUSP6 mRNA水平;MTT法检测细胞增殖情况;流式细胞术检测细胞凋亡率;划痕实验检测细胞迁移能力;Western blot检测细胞中DUSP6、细胞外信号调节激酶1/2(ERK1/2)、p-ERK1/2蛋白表达。结果:与空白对照组和过表达NC组比较,过表达组DUSP6 mRNA相对表达量升高,MMT法检测24、48、72 h细胞增殖能力下降,细胞凋亡率升高,愈合率降低,DUSP6蛋白相对表达量升高,p-ERK1/2蛋白相对表达量降低(P<0.05);与空白对照组和沉默NC组比较,沉默组DUSP6 mRNA相对表达量降低,MTT法24、48、72 h增殖能力增加,细胞凋亡率降低,愈合率升高,DUSP6蛋白相对表达量降低,p-ERK1/2蛋白相对表达量升高(P<0.05);过表达组DUSP6 mRNA及蛋白相对表达量高于沉默组,24、48、72 h细胞增殖能力小于沉默组,细胞凋亡率高于沉默组,愈合率低于沉默组,p-ERK1/2蛋白相对表达量低于沉默组(P<0.05)。结论:DUSP6沉默可促进人子宫内膜癌Ishikawa细胞增殖和迁移,抑制细胞凋亡,可能是通过调控ERK1/2通路发挥作用。

Objective:To investigate the effect and mechanism of dual specificity phosphatase 6(DUSP6)silencing on the apoptosis and migration of human endometrial cancer Ishikawa cells.Methods:Ishikawa cells were randomly divided into blank control group(transfection liposome reagent),overexpression group(transfection DUSP6),overexpression NC group(transfection DUSP6-NC),silent group(transfection siDUSP6),silent NC Group(transfection siDUSP6-NC). RT-qPCR was used to detect DUSP6 mRNA level in cells,MTT method was used to detect cell proliferation,flow cytometry was used to detect cell apoptosis rates,scratch test was used to detect cell migration ability,Western blot was used to detect DUSP6 and extracellular signal-regulated kinase1/2 in cells(ERK1/2),p-ERK1/2 protein expression.Results:Compared with the blank control group and the overexpression NC group,the relative expression of DUSP6 mRNA increased,the MTT method 24,48,72 h cell multiplication capacity decreased,the apoptosis rate increased,the healing rate decreased,and the relative expression level DUSP6 protein increased,the relative expression level of p-ERK1/2 protein decreased in the overexpression group increased(P<0.05). Compared with the blank control group and the silent NC group,the silence group DUSP6 mRNA relative expression level decreased,and 24,48,72 h cell multiplication capacity increased,cell apoptosis rate decreased,healing rate increased,the relative expression of DUSP6 protein decreased,and the relative expression of p-ERK1/2 protein increased(P<0.05). The relative expression of DUSP6 mRNA and protein in the overexpression group was higher than that of silence group,the 24,48 and 72 h multiplication capacity of the MTT method was lower than the silent group,the apoptosis rate was higher than that of the silent group,the healing rate was lower than that of the silent group,and the relative expression of p-ERK1/2 protein was lower than that of the silent group(P<0.05).Conclusion:Silencing DUSP6 can promote the proliferation and migration of human endometrial cancer Ishikawa cells,and inhibit cell apoptosis,possibly by regulating the ERK1/2 pathway.