【摘要】 目的探究内源性干扰双调蛋白(AREG)的表达对卵巢癌细胞迁移与侵袭及化疗敏感性的影响。方法构建靶向AREG-siRNA干扰质粒并转染卵巢癌细胞COC1,此为AREG转染组,同时设立正常对照组和阴性转染组。采用MTT法检测各组COC1的体外生长抑制率及对顺铂化疗的敏感性,细胞划痕实验及Transwell侵袭实验探索AREG对COC1迁移及侵袭行为的影响,实时荧光定量PCR检测各组细胞中AREG mRNA的表达量,Western blot法检测各组细胞中AREG、表皮生长因子受体(EGFR)及细胞外信号调节激酶(ERK)蛋白表达水平。结果与正常对照组及阴性转染组相比较,AREG转染组细胞的增殖受到抑制,随着作用时间的延长,COC1生长抑制率显著上升(P<0.01),且AREG转染组细胞的侵袭及迁移也被抑制(P<0.05);AREG转染组细胞对顺铂药物的化疗敏感性增强(P<0.05)。与正常对照组及阴性转染组相比,AREG转染组细胞中AREG mRNA的表达量和AREG、EGFR及ERK蛋白表达水平显著下降,差异有统计学意义(P<0.05)。结论 AREG可显著抑制卵巢癌细胞COC1的体外生长、迁移和侵袭,提高卵巢癌细胞的化疗敏感性,推测其可能的作用机制与AREG参与调控EGFR-ERK信号通路相关蛋白的表达有关。更多还原

【Abstract】 Objective:To investigate the effects of the expression of endogenous interfering amphiregulin on migration,invasion and chemosensitivity of ovarian cancer cells.Methods:AREG-siRNA targeting plasmid was constructed and transfected to ovarian cancer cell COC1 in the AREG transfection group. A normal control group and a negative transfection group were set up at the same time. The growth inhibition rate of COC1 in vitro in each group and the sensitivity to cisplatin chemotherapy were detected by MTT assay. The effects of amphiregulin on the migration and invasion of COC1 were investigated by cell scratch assay and Transwell invasion assay. AREG mRNA expression of cells in each group was detected by real-time fluorescence quantitative PCR. Western blot was used to detect the expressions of proteins of AREG,epidermal growth factor receptor(EGFR)and extracellular signal-regulated kinase(ERK)of cells in each group.Results:Compared with the normal control group and the negative transfection group,the proliferation of the cells in the AREG transfection group was significantly inhibited,and the growth inhibition rate of COC1 increased with the prolongation of the action time(P<0.01). The invasion and migration of AREG transfected cells were also significantly inhibited. The sensitivity to cisplatin chemotherapy of AREG transfection group was significantly enhanced(P<0.05). The mRNA expression of AREG mRNA and the protein expression of AREG,EGFR and ERK in AREG transfected group were significantly lower than those in normal control group and negative transfection group(P<0.05).Conclusions:AREG can significantly inhibit the growth,migration and invasion of ovarian cancer cells COC1 in vitro,and significantly improve the sensitivity of ovarian cancer cells to chemosensitivity. Its mechanism may be related to the involvement of AREG in regulating the expression of EGFR-ERK signaling pathway-related proteins.更多还原