RASAL3通过上调FXYD6表达促进肝内胆管细胞癌细胞增殖的作用机制研究

RASAL3 promotes the proliferation of intrahepatic cholangiocarcinoma cells by up regulating the expression of FXYD6

目的探讨Ras蛋白激活类似物3(RASAL3)在肝内胆管癌(iCCA)中的表达并深入研究其促进iCCA进展的作用机制。方法收集185例iCCA患者肿瘤与癌旁组织标本,应用免疫组化、RT-qPCR和Western blot法检测RASAL3的表达,并同时检测人胆管癌细胞株和人胆管上皮细胞中RASAL3和FXYD6的mRNA和蛋白表达情况,CCK-8法检测细胞增殖及Na^(+)-K^(+)-ATP酶活性。结果 RASAL3在胆管癌组织和细胞株中高表达;生存分析结果显示RASAL3高表达与胆管癌的不良预后相关(P<0.05)。敲降RASAL3抑制胆管癌细胞的增殖;沉默RASAL3降低FXYD6表达水平,抑制Na^(+)-K^(+)-ATP酶活性。结论 iCCA中RASAL3表达水平显著上调,RASAL3可通过激活FXYD6,上调Na^(+)-K^(+)-ATP酶活性促进iCCA的发展。

Objective To investigate the expression of RASAL3 in intrahepatic cholangiocarcinoma(iCCA)and the mechanism of promoting iCCA development.Methods Tumor and paracancerous tissues were collected from 185 iCCA patients,the expression of RASAL3 was detected by immunohistochemistry,RT-qPCR and Western blot.The expression of RASAL3 and FXYD6 mRNA and protein in human cholangiocarcinoma cell line and human bile duct epithelial cells were detected with RT-qPCR and Western blot,the cell proliferation was detected with CCK-8 assay,and the activity of Na^(+)-K^(+)-ATPase was also detected.Results RASAL3 was highly expressed in cholangiocarcinoma tissues and cell lines;Survival analysis showed that RASAL3 overexpression was associated with poor prognosis of cholangiocarcinoma(P<0.05)and knockdown of RASAL3 inhibits the proliferation of cholangiocarcinoma cells;Silencing RASAL3 decreases the expression of FXYD6 inhibiting the activity of Na^(+)-K^(+)-ATPase.Conclusion RASAL3 is up-regulated in human cholangiocarcinoma,which can promote the occurrence and development of cholangiocarcinoma by activating FXYD6 and affecting Na^(+)-K^(+)-ATPase activity.