TP53基因沉默介导PI3K/PTEN/AKT信号通路对肾透明细胞癌侵袭转移的调控机制

目的探讨肿瘤蛋白p53基因(TP53)沉默介导PI3K/PTEN/AKT信号通路对肾透明细胞癌侵袭转移的调控机制。方法取60例肾透明细胞癌组织标本,免疫组化分析TP53蛋白在肾透明细胞癌组织中的表达;取人肾透明细胞癌细胞株RLC-310进行细胞培养;细胞转染表达载体分为空白组(A组)、阴性对照组(B组)、sh TP53组(C组)、PTEN抑制剂bpv组(D组)、phen+sh TP53组(E组)。采用实时荧光定量PCR(qRT-PCR)和Western Blot检测各组细胞TP53、同源性磷酸酶-张力蛋白(PTEN)、磷脂酰肌醇-3-激酶(PI3K)、AKT的m RNA和蛋白表达水平;CCK-8法检测各组细胞增殖能力;划痕愈合试验检测各组细胞的迁移能力;Transwell侵袭试验检测各组细胞的侵袭能力。结果TP53蛋白在肾透明细胞癌组织中高表达(P<0.05);与A组和B组相比,C组的TP53,PI3K,AKT m RNA和蛋白表达水平显著下降,PTEN m RNA和蛋白表达水平均显著上升,且细胞增殖能力、迁移率、侵袭率均显著下降(P<0.05);D组TP53,PI3K,AKT m RNA和蛋白表达水平显著上升,PTEN m RNA和蛋白表达水平显著下降,且细胞增殖能力、迁移率、侵袭率均显著上升(P<0.05);同时,E组TP53下降(P<0.05),而其他指标与A组和B组相当(P>0.05)。结论沉默TP53基因抑制PI3K/PTEN/AKT信号通路,从而抑制肾透明细胞癌细胞浸润转移功能,并可逆转phen诱导的肾透明细胞癌细胞浸润转移。

Regulation Mechanism of TP53 Gene Silencing Mediating PI3K/PTEN/AKT Signaling Pathway on Invasion and Metastasis of Renal Clear Cell Carcinoma

Objective To explore the regulation mechanism of tumor protein p53(TP53)gene silencing mediating PI3 K/PTEN/AKT signaling pathway on invasion and metastasis of renal clear cell carcinoma.Methods A total of 60 cases of renal clear cell carcinoma was collected,and immunohistochemistry was used to analyze the expression of TP53 protein.Human renal clear cell carcinoma cell line RLC-310 was cultured and transfected into five groups:Blank group(group A),negative control(NC,transfection of short hairpin RNA NC)group(group B),sh TP53 group(group C),PTEN inhibitor bpv(phen)group(group D)and phen+shTP53 group(group E).The expression levels of TP53,phosphatase and tensin homolog(PTEN),phosphatidylinositol-3-kinase(PI3 K)and AKT were detected by Quantitative real-time PCR(q RT-PCR)and Western blot;the proliferation ability of cells was detected by CCK-8 method;the migration ability of cells in each group was detected by scratch healing test;and the invasion ability of cells in each group was detected by Transwell invasion test.Results Clinical experiment showed that TP53 protein was highly expressed in renal clear cell carcinoma(P<0.05).Cell experiment showed that compared with group A and group B,the levels of TP53,PI3 K and AKT m RNA and protein expression in group C were significantly down-regulated,and the levels of PTEN m RNA and protein expression were significantly up-regulated,and cell proliferation,migration and invasion were significantly down-regulated(P<0.05);the levels of TP53,PI3 K and AKT m RNA and protein expression in group D were significantly up-regulated,while the levels of PTEN m RNA and protein expression were significantly down-regulated,and the cell proliferation,migration and invasion were significantly up-regulated(P<0.05);meanwhile,TP53 was down-regulated in group E(P<0.05),but there was no significant difference in other indicators compared with group A and group B(P>0.05).Conclusion TP53 gene silencing inhibits PI3 K/PTEN/AKT signaling pathway,thus inhibits the infiltration and metastasis of renal clear cell carcinoma cells,and reverses PHEN-induced infiltration and metastasis of renal clear cell carcinoma cells.