抑制YAP对宫颈癌细胞增殖、凋亡、迁移及p53、Fra-1表达的影响

Inhibition of YAP on proliferation,apoptosis,migration and expression of p53 and FRA-1 in cervical cancer cells

目的探讨抑制YAP对宫颈癌细胞增殖、凋亡、迁移及p53、Fra-1表达的影响。方法宫颈癌细胞株随机分为宫颈癌组(宫颈癌细胞无转染),NC组(宫颈癌细胞转染YAP-shRNA空载体质粒NC),和YAP-shRNA组(宫颈癌细胞转染YAP-shRNA),RT-PCR检测YAP基因表达;CCK8检测Hela细胞增殖;流式细胞仪检测Hela细胞凋亡率;Transwell小室检测Hela细胞迁移数目;免疫印迹及PCR分别检测p53、Fra-1表达。结果 NC组与宫颈癌组Hela细胞株中YAP基因水平比较差异无统计学意义(P>0.05),YAP-shRNA组中Hela细胞株中YAP基因水平均低于NC组与宫颈癌组(P<0.05);宫颈癌组和NC组Hela细胞吸光度(A)值在24、48、72 h比较差异无统计学意义(P>0.05),YAP-shRNA组Hela细胞吸光度(A)值均低于宫颈癌组和NC组(P<0.05);宫颈癌组和NC组Hela细胞凋亡率、迁移数目差异无统计学意义(P>0.05),与宫颈癌组和NC组比较,YAP-shRNA组凋亡率升高、迁移数目降低(P<0.05);宫颈癌组和NC组Hela细胞p53、Fra-1蛋白及mRNA水平比较差异无统计学意义(P>0.05),YAP-shRNA组p53、Fra-1蛋白及mRNA水平升高,与宫颈癌组和NC组差异有统计学意义(P<0.05)。结论 YAP在宫颈癌细胞中存在高表达,沉默后能够降低宫颈癌Hela细胞株增殖及迁移能力,加快细胞凋亡率,这可能与激活P53及Fra-1基因相关。

Objective To investigate the effects of inhibition of YAP on proliferation, apoptosis, migration, p53 and FRA-1 expression of cervical cancer.Methods Cervical cancer cells were randomly divided into cervical cancer group(cervical cancer cells without transfection),NC group(cervical cancer cells transfected with empty plasmid),and YAP-shRNA group(cervical cancer cells transfected with YAP-shRNA).The expression of YAP gene was detected by RT-PCR.CCK8 was used to detect cell proliferation.Flow cytometry was used to detect apoptosis.The cell migration was detected by Transwell chamber.The expressions of p53 and FRA-1 were detected by Western blot and PCR respectively.Results There was no significant difference in YAP gene level between NC and cervical cancer groups, but YAP gene level in YAP-shRNA group was lower than NC and cervical cancer groups(P<0.05).There was no difference in Hela cell absorbance(A) between cervical cancer and NC groups at 24,48 and 72 h, but the Hela cell absorbance(A) in YAP-shRNA group was lower than that in cervical cancer and NC groups(P<0.05).There were no significant differences in the apoptosis rate and migration number of Hela cells between cervical cancer and NC groups, but the apoptosis rate and migration number of YAP-shRNA group were increased and decreased, respectively, and the differences were significant(P<0.05).There was no difference in p53 and FRA-1 protein and mRNA levels between cervical cancer and NC groups, which were increased in YAP-shRNA group(P<0.05).Conclusion There is high expression of YAP in cervical cancer cells.Silencing YAP can reduce the proliferation and migration ability of cervical cancer Hela cells and accelerate the apoptosis rate, which may be related to the activation of p53 and FRA-1 gene/proteins.