慢病毒介导HMGN5基因沉默对卵巢癌细胞增殖、迁移、侵袭的影响及其机制研究

Effect of lentivirus mediated HMGN5 gene silencing on proliferation,migration and invasion of ovarian cancer cells and its mechanism

目的观察慢病毒介导的高迁移率族核小体结合域5(HMGN5)基因沉默对卵巢癌细胞增殖、迁移和侵袭的影响,并对其机制进行初步探讨。方法RT qPCR法检测人卵巢上皮细胞株IOSE及4株不同卵巢癌细胞中HMGN5的表达情况,将HMGN5 shRNA慢病毒载体转染至人卵巢癌细胞SKOV3中,RT qPCR和Western blot检测SKOV3细胞中HMGN5 mRNA和蛋白表达情况,MTT法检测沉默HMGN5基因对细胞增殖能力的影响,划痕实验检测沉默HMGN5基因对细胞迁移能力的影响,Tran swell实验检测沉默HMGN5基因对细胞侵袭能力;Western blot检测SKOV3细胞中PCNA、MMP 9、Wnt1和β 连环蛋白(β catenin)蛋白表达水平。结果与卵巢上皮细胞株IOSE相比,HMGN5在卵巢癌细胞株SKOV3、A2780、HO 8910、Caov3中的表达明显升高(P<0.05)。与Blank组和NC组相比,sh HMGN5组SKOV3细胞中HMGN5 mRNA和蛋白表达水平显著降低(P<0.05),SKOV3细胞增殖、迁移和侵袭能力均明显下降(P<0.05),细胞中PCNA、MMP 9、Wnt1和β catenin蛋白表达水平明显下调(P<0.05)。结论沉默HMGN5可能通过阻碍Wnt/β catenin信号通路的激活抑制人卵巢癌SKOV3细胞增殖、迁移和侵袭能力。

Objective To investigate the effects of lentiviral mediated high mobility group nucleosome binding domain 5(HMGN5)gene silencing on proliferation,migration and invasion of ovarian cancer cells,and to explore the mechanism.Methods RT qPCR was used to detect the expression of HMGN5 in human ovarian epithelial cell line IOSE and four different ovarian cancer cells.HMGN5 shRNA lentivirus vector was transfected into human ovarian cancer cell line SKOV3.RT qPCR and Western blot were used to detect the expression of HMGN5 mRNA and protein in SKOV3 cells.MTT method was used to detect the effect of HMGN5 gene silencing on cell proliferation,scratch test was used to detect the effect of HMGN5 gene silencing on cell migration,and Transwell test was used to detect the invasion of HMGN5 gene silencing on cell.Western blot was used to detect the expression of PCNA,MMP 9,Wnt1 andβ Catenin in SKOV3 cells.Results The expression of HMGN5 in SKOV3,A2780,HO 8910 and CAOV3 was significantly higher than that in IOSE(P<0.05).Compared with blank group and NC group,the expression level of HMGN5 mRNA and protein in SKOV3 cells of sh HMGN5 group was significantly lower(P<0.05),the proliferation,migration and invasion ability of SKOV3 cells were significantly lower(P<0.05),and the expression levels of PCNA,MMP 9,Wnt1 andβ Catenin cells were significantly lower(P<0.05).Conclusion Silencing HMGN5 may inhibit the proliferation,migration and invasion of human ovarian cancer SKOV3 cells by blocking the activation of Wnt/β catenin signaling pathway.