PTEN通过靶向P21诱导卵巢癌细胞衰老的机制研究

PTEN induces ovarian cancer cell senescence by targeting P21

目的:探讨抑癌基因PTEN靶向衰老蛋白P21在促进卵巢癌细胞衰老中的作用及相关机制。方法:收集30例人卵巢癌组织样本和30例卵巢囊肿卵巢样本,Real-time PCR、Western blot及免疫荧光染色检测PTEN、P21和TRIM39的表达;体外培养SKOV3卵巢癌细胞,分别在SKOV3细胞中过表达P21和PTEN以及干扰TRIM39,运用Real-time PCR和Western blot分别检测PTEN、P21及TRIM39的表达,β-gal染色鉴定衰老细胞,Ki67染色检测细胞增殖。结果:与人正常卵巢组织相比,人卵巢癌中PTEN和P21的表达显著减少(P<0.01);在SKOV3细胞中过表达P21和PTEN能显著促进细胞衰老(P<0.01),抑制细胞增殖(P<0.01),在SKOV3细胞中过表达P21并不影响PTEN的表达水平,而在SKOV3细胞中分别过表达PTEN和TRIM39都显著促进了P21的表达(P<0.01);在SKOV3细胞中过表达TRIM39同样促进了细胞衰老(P <0.01),PTEN激动剂则能够促进SKOV3细胞中TRIM39的表达(P<0.01);干扰SKOV3细胞中TRIM39的表达则抑制了PTEN激动剂促进P21表达的作用(P<0.01),同样抑制了细胞衰老。结论:PTEN通过促进TRIM39表达维持了P21的稳定,减少了P21的降解,从而促进了SKOV3细胞衰老,抑制了卵巢癌的发生发展。

Objective:To investigate the role of PTEN targeted senescence protein P21 in promoting the senescence of ovarian cancer cells and its related mechanism.Methods:30 samples of human ovarian cancer and 30 samples of ovarian cyst were collected,the expressions of PTEN,P21 and TRIM39 were detected by Real-time PCR,Western blot and immunofluorescence staining. SKOV3 ovarian cancer cells were cultured in vitro,overexpression of P21 and PTEN or interference with TRIM39 were performed in SKOV3 cells. Real-time PCR and Western blot were used to detect the expression of PTEN,P21 and TRIM39;β-gal staining was used to identify senescent cells,and Ki67 staining was used to detect cell proliferation.Results:Compared with normal ovarian tissue,the expression of PTEN and P21 in human ovarian cancer tissue decreased significantly(P<0.01). Overexpression of P21 and PTEN in SKOV3 cells significantly promoted cell senescence(P<0.01)and inhibited cell proliferation(P< 0.01). Overexpression of P21 did not affect the expression of PTEN in SKOV3 cells,overexpression of PTEN and TRIM39 respectively in SKOV3 cells significantly promoted the expression of P21(P<0.01). Overexpression of TRIM39 in SKOV3 cells also promoted cell senescence(P<0.01),and PTEN agonist also promoted the expression of TRIM39 in SKOV3 cells(P<0.01). Interference with TRIM39 in SKOV3 cells inhibited the promotion role of PTEN agonist on P21 expression(P<0.01)and also inhibitsed cell senescence.Conclusion:PTEN maintains the stability of P21 and reduces the degradation of P21 by promoting the expression of TRIM39,thus promoting the senescence of SKOV3 cells and inhibiting the occurrence and development of ovarian cancer.