USP7在结直肠癌组织中的表达及其通过调控Wnt/PCP通路对细胞增殖、凋亡的影响

USP7 expression in colorectal cancer tissues and its effect on cell proliferation and apoptosis by regulating Wnt/PCP pathway

目的:探讨泛素特异性蛋白酶7(ubiquitin specific protease 7,USP7)在结直肠癌组织中的表达及其通过调控Wnt/PCP通路对结直肠癌细胞SW480增殖、凋亡的影响和机制研究。方法:免疫组织化学法检测USP7在结直肠癌组织中的表达;qRT-PCR和Western blotting检测USP7在结直肠癌组织和结直肠癌细胞系中的表达;qRT-PCR和Western blotting检测沉默USP7在SW480细胞中的表达;细胞克隆形成实验检测SW480细胞增殖能力;Transwell迁移实验检测SW480细胞迁移能力;流式细胞术检测SW480细胞的凋亡情况;Western blotting检测Wnt/PCP通路相关蛋白的表达。结果:USP7在结直肠癌组织和结直肠癌细胞系中高表达(P<0.05);沉默USP7使SW480细胞中USP7的表达降低(P<0.001),细胞的增殖和迁移能力下降(P<0.001),细胞凋亡率增加(P<0.001);与shN C组相比,沉默USP7使Wnt7a蛋白和RhoA蛋白的表达水平显著下调(P<0.001,P<0.001),JNK蛋白磷酸化程度显著降低(P<0.01)。结论:USP7在结直肠癌组织和结直肠癌细胞系中高表达。沉默USP7抑制SW480细胞增殖、迁移并诱导细胞凋亡,可能与Wnt/PCP通路有关。

Objective:To investigate the effect and mechanism of USP7 expression in colorectal cancer and its regulation of Wnt/PCP pathway on proliferation and apoptosis of colorectal cancer cell SW480.Methods:Immunohistochemistry was used to detect the expression of USP7 in colorectal cancer tissues.qRT-PCR and Western blotting were used to detect the expression of USP7 in colorectal cancer tissues and colorectal cancer cell lines.qRT-PCR and Western blotting were used to detect the expression of silent USP7 in SW480 cells.Cell clone formation experiment was used to test SW480 cell proliferation ability.Transwell migration test was used to detect the migration ability of SW480 cells.Cell flow cytometry was uesd to detect the apoptosis of SW480 cells.Western blotting was used to detect the expression of Wnt/PCP pathway related proteins.Results:USP7 was highly expressed in colorectal cancer tissues and colorectal cancer cell lines(P<0.05).Silencing USP7 reduced the expression of USP7 in SW480 cells(P<0.001),decreased the cell proliferation and migration ability(P<0.001),and increased the apoptosis rate(P<0.001).Compared with the shNC group,silencing USP7 significantly reduced the expression levels of Wnt7a protein and RhoA protein(P<0.001,P<0.001),and the phosphorylation degree of JNK protein was significantly reduced(P<0.01).Conclusion:USP7 is highly express in colorectal cancer tissues and colorectal cancer cell lines.Silencing USP7 inhibit SW480 cell proliferation,migration and induces apoptosis,which may be related to the Wnt/PCP pathway.