circRNA-ITCH通过TET1基因抑制Wnt/β-catenin信号通路对鼻咽癌细胞增殖和侵袭的影响

CircRNA-ITCH Affects Proliferation and Invasion of Nasopharyngeal Carcinoma Cells by Inhibiting Wnt/β-catenin Signaling Pathway through TET1 Gene

目的观察环状RNA(circRNA)-ITCH在鼻咽癌组织和细胞系中的表达,探讨circRNA-ITCH对鼻咽癌细胞增殖和侵袭的影响及其分子机制。方法qPCR分别检测circRNA-ITCH在49例鼻咽癌组织和鼻咽癌细胞系中的表达。选取circRNA-ITCH表达最低的细胞系,分别转染阴性质粒或过表达circRNA-ITCH的质粒,定义为对照组和实验组。CCK-8法和Transwell侵袭实验分别检测过表达circRNA-ITCH对细胞增殖和侵袭能力的影响。qPCR和Western blot法分别检测过表达circRNA-ITCH对10-11转位酶1(ten-eleven translocation1,TET1)基因和Wnt/β-catenin信号通路表达的影响。结果circRNA-ITCH在鼻咽癌组织中表达水平明显低于癌旁组织(P<0.01)。circRNA-ITCH在鼻咽癌细胞系中表达水平明显低于人永生化鼻咽上皮细胞(P<0.05),在CNE-2Z细胞中的表达水平最低(P<0.01)。与对照组比较,实验组CNE-2Z细胞的增殖能力明显降低(P<0.05),细胞侵袭能力明显降低(P<0.01)。与对照组比较,实验组CNE-2Z细胞中TET1在mRNA和蛋白水平的表达明显增加(P<0.01),Wnt1、PLC、PKC和β-catenin蛋白表达明显降低,Wnt/β-catenin信号通路被抑制。结论circRNA-ITCH在鼻咽癌组织及细胞系中表达降低,过表达circRNA-ITCH可明显抑制鼻咽癌CNE-2Z细胞的增殖和侵袭能力,其分子机制可能是促进TET1基因的表达,抑制Wnt/β-catenin信号通路。

Objective To observe the expression of circRNA-ITCH in nasopharyngeal carcinoma tissues and cell lines,and to explore the effect and molecular mechanism of circRNA-ITCH on the proliferation and invasion of nasopharyngeal carcinoma cells.Methods qPCR was used to detect the expression of circRNA-ITCH in 49 nasopharyngeal carcinoma tissues and nasopharyngeal carcinoma cell lines.Cell lines with the lowest circRNA-ITCH expression were selected and transfected with negative plasmids or plasmids that over-expressed circRNA-ITCH,respectively,and defined as control and experimental groups.CCK-8 method and Transwell invasion test were used to detect the effects of over-expressed circRNA-ITCH on cell proliferation and invasive ability.qPCR and Western blotting were used to detect the effects of circRNA-ITCH overexpression on the expression of ten-eleven translocation1(TET1)gene and Wnt/β-catenin signaling pathway.Results The expression level of circRNA-ITCH in nasopharyngeal carcinoma tissues was significantly lower than that in adjacent tissues(P<0.01).The expression level of circRNA-ITCH in nasopharyngeal carcinoma cell lines was significantly lower than that of human immortalized nasopharyngeal epithelial cells(P<0.05),and the expression level in CNE-2Z cells was the lowest(P<0.01).Compared with the control group,the proliferation ability of CNE-2Z cells in the circRNA-ITCH group was significantly reduced(P<0.05),and the cell invasion capacity was significantly reduced(P<0.01).Compared with the control group,the expression of TET1 at the mRNA and protein levels in the cells of the circRNA-ITCH group were significantly increased(P<0.01),Wnt1,PLC,PKC,andβ-catenin proteins expression were significantly reduced,and the Wnt/β-catenin signaling pathway was significantly reduced.Conclusion circRNA-ITCH expression is reduced in nasopharyngeal carcinoma tissues and cell lines.Overexpression of circRNA-ITCH can significantly inhibit the proliferation and invasion of nasopharyngeal carcinoma CNE-2Z cells.Its molecular mechanism may be to promote the expression of TET1 gene and inhibit Wnt/β-catenin signaling pathway.