肝细胞癌中CAⅨ表达与侵袭转移的关系

Expression of CAⅨin hepatocellular carcinoma and its effect on the proliferation,invasion and metastasis of hepatoma cells

目的探讨碳酸酐酶Ⅸ(carbonic anhydraseⅨ,CAⅨ)在肝细胞癌和癌旁组织中的表达及与肝细胞癌侵袭、转移的关系。方法采用免疫组化LDP法检测67例肝细胞癌及癌旁组织中CAⅨ的表达,并分析其与临床病理特征及预后的关系。将CAⅨ过表达载体及干扰表达的siRNA分别转染至人肝癌细胞株HepG2后,应用qRT-PCR法检测各组CAⅨ的表达,CCK-8实验检测CAⅨ对肝癌细胞增殖能力的影响,细胞迁移/侵袭实验检测其对肝癌细胞侵袭转移能力的影响。从TCGA数据库中检索肝细胞癌CAⅨ表达与预后的资料,进行对比分析。结果CAⅨ在肝细胞癌中的表达显著高于癌旁组织(P<0.05),且随着肝细胞癌分化程度的降低其阳性率逐渐升高(P<0.05)。随访中位时间17个月,Kaplan-Meier生存曲线显示,CAⅨ阳性者和阴性者的总生存率差异有统计学意义(P<0.05)。TCGA数据库中CAⅨmiRNA高表达的肝细胞癌患者总生存率显著低于低/中等表达者(P=0.00045)。细胞增殖实验及细胞迁移/侵袭实验显示,CAⅨ过表达能促进肝癌细胞HepG2的增殖及侵袭转移能力,siRNA干扰沉默HepG2细胞中CAⅨ的表达抑制其增殖及侵袭转移能力(P<0.05)。结论CAⅨ表达升高与肝细胞癌组织分化程度密切相关,且提示预后较差,其在肝癌组织的高表达可能具有促进肝癌细胞增殖及侵袭转移的能力。

Purpose To investigate the expression of carbonic anhydraseⅨ(CAⅨ)in hepatocellular carcinoma(HCC)and corresponding para-cancer tissues,and to explore its relationship with invasion and metastasis of hepatocellular carcinoma.Methods Immunohistochemistry(IHC)was used to examine the expression of CAⅨin 67 samples of HCC tissues and corresponding para-cancer tissues and to analysis the correlation between the expression of CAⅨwith clinicopathologic features and prognosis.After CAⅨoverexpression vector and interfering siRNA were transfected into human hepatoma cell line HepG2,the expression of CAⅨin each group was detected by qRT-PCR analysis.CCK-8 assay was used to detect the cell proliferation.The effect of CAⅨon the invasion and metastasis of HepG2 cells was detected by cell migration/invasion assay.The data of the expression of CAⅨand prognosis in hepatocellular carcinoma were extracted from The Cancer Genome Atlas(TCGA)database and make a comparative analysis.Results The expression of CAⅨin hepatocellular carcinoma was significantly higher than that in corresponding para-cancer tissues(P<0.05).The expression level of CAⅨwas statistically significantly different between different type of differentiation(P<0.05).The median follow-up time was 17 months.Kaplan-Meier survival curve showed there was statistically significant differences in overall survival rates between CAⅨpositive and CAⅨnegative patients(P<0.05).In TCGA database,the overall survival rate of hepatocellular carcinoma patients with high miRNA expression of CAⅨshowed significantly lower than that of patients with low+medium expression of CAⅨ(P=0.00045).Cell proliferation assay and cell migration/invasion assay showed that CAⅨoverexpression could promote the proliferation,invasion and metastasis of HepG2 cells.siRNA interference silenced the expression of CAⅨcells to inhibit the proliferation,invasion and metastasis of HepG2 cells(P<0.05).Conclusion The increased expression of CAⅨis closely related to the differentiation of hepatocellular carcinoma,and suggests that the prognosis is poor.The overexpression of CAⅨgene in hepatocellular carcinoma may enhance the proliferation,invasion and metastasis of hepatocellular carcinoma cells.