JOSD1在肝细胞癌中的表达及对细胞增殖、克隆、迁移的影响研究

Expression of JOSD1 in hepatocellular carcinoma and its effects on cell proliferation,clonogenicity,and migration

目的探讨含约瑟芬结构域的蛋白1(JOSD1)在肝细胞癌(HCC)中的表达及对HCC细胞增殖、克隆、迁移的影响。方法利用公共生物信息学分析平台(http://gepia.cancer-pku.cn/detail.php)分析JOSD1在HCC组织及癌旁组织的表达差异;收集2022年10月至2023年5月在嘉兴市第一医院行肝癌根治术的4例HCC组织及癌旁组织标本,分析两种组织中的JOSD1蛋白表达差异;选择人HCC细胞株(Huh7),转染特异性小干扰RNA(siRNA),并设立JOSD1敲低组(siJOSD1组)和阴性对照组(siNC组);CCK-8法、克隆形成实验、Transwell迁移实验检测敲低JOSD1对Huh7细胞增殖、克隆、迁移能力的影响;Western blot法检测敲低JOSD1对Huh7细胞E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)表达水平的影响。结果公共生物信息学分析显示HCC组织中JOSD1表达水平明显高于癌旁组织(P<0.05),进一步临床样本验证发现,HCC组织JOSD1蛋白表达水平明显高于癌旁组织(P<0.05);与siNC组相比,siJOSD1组Huh7细胞的增殖、克隆、迁移能力均下降(均P<0.05),E-cadherin蛋白表达水平上调(P<0.05),N-cadherin、Vimentin蛋白表达水平均下调(均P<0.05)。结论HCC中JOSD1表达上调,敲低JOSD1可显著降低HCC细胞的增殖、克隆、迁移能力,可能通过上调E-cadherin,下调N-cadherin、Vimentin蛋白表达水平发挥作用。

Objective To explore the expression of deubiquitinase Josephin domain-containing protein 1(JOSD1)in hepatocellular carcinoma(HCC)and its impact on proliferation,cloning,and migration of HCC cells.Methods The public bioinformatics analysis platform Gene Expression Profiling Interactive Analysis(GEPIA,http://gepia.cancer-pku.cn/detail.php)was used to analyze the differential expression of JOSD1 in HCC tissues and adjacent tissues;Four specimens of HCC tissues and normal adjacent tissues were collected from patients who received HCC radical resection in the First Hospital of Jiaxing from October 2022 to May 2023,and the differences in JOSD1 protein expression between liver cancer tissue and adjacent tissues were further analyzed.Human HCC cell line(Huh7)was selected to construct JOSD1 knockdown group(siJOSD1 group)and negative control group(siNC group)using small interfering RNA(siRNA)transfection technology.The effects of JOSD1 knockdown on the proliferation,cloning,and migration ability of Huh7 cells were assessed using the CCK-8 method for cell proliferation assay,clone formation assay,and Transwell migration assay.Additionally,the expression levels of E-cadherin,N-cadherin,and vimentin in Huh7 cells were determined through Western blotting analysis to evaluate the impact of JOSD1 knockdown on them.Results The analysis based on GEPIA data platform showed that the expression level of JOSD1 in HCC tissue was significantly higher than that of normal adjacent tissue(P<0.05).Further clinical sample validation showed that the expression level of JOSD1 protein in HCC tissue was significantly higher than that of adjacent tissue(P<0.05).Compared with the siNC group,knocking down JOSD1 significantly reduced the proliferation,cloning,and migration ability of Huh7 cells,significantly upregulated the expression of E-cadherin protein in liver cancer cells,and downregulated the expressions of N-cadherin and Vimentin proteins(all P<0.05).Conclusion JOSD1 expression is upregulated in HCC,while knocking down can significantly reduce the proliferation,cloning and migration ability of HCC cells,possibly by upregulating E-cadherin and downregulating N-cadherin and Vimentin proteins.