【摘要】 目的探讨肿瘤相关钙信号转导蛋白2(Trop-2)基因对肝癌发生发展的影响及作用机制。方法收集10例肝癌患者的肝癌组织标本和癌旁正常组织标本,免疫组织化学染色法和实时荧光定量聚合酶链反应(qRT-PCR)法分别检测不同组织中Trop-2蛋白和mRNA的表达情况;流式细胞术和蛋白质印迹法(Western blot)检测HepG2、HCCLM3、HL-7702细胞中Trop-2蛋白的表达情况;Western blot法检测不同组织和转染后HepG2细胞中Trop-2、蛋白激酶C-α(PKC-α)、磷酸化蛋白激酶C-α(p-PKC-α)和核因子-κB(NF-κB)的表达情况;CCK-8法检测转染后HepG2细胞的增殖率。结果肝癌组织中p-PKC-α、NF-κB、Trop-2蛋白及Trop-2 mRNA的相对表达量均明显高于癌旁正常组织(P﹤0.01);HCCLM3细胞和HepG2细胞中Trop-2蛋白的相对表达量均明显高于HL-7702细胞(P﹤0.01);Trop-2 siRNA组HepG2细胞中Trop-2、p-PKC-α、PKC-α和NF-κB蛋白的相对表达量均低于空白对照组和阴性对照组(P﹤0.05);Trop-2 siRNA组HepG2细胞的增殖率低于阴性对照组(P﹤0.05)。结论Trop-2基因在肝癌组织和肝癌细胞株中均有较高水平的表达,提示其可能是肝癌潜在的治疗靶点。更多还原

【Abstract】 Objective To investigate the impact of tumor-associated calcium signal transducer 2(Trop-2) gene on the occurrence and development of hepatic carcinoma and the underlying mechanism. Method Samples of hepatic carcinoma tissues and adjacent normal tissues were collected from 10 patients with hepatic carcinoma. Immunohistochemical staining and quantitative real-time polymerase chain reaction(qRT-PCR) were used to detect the expression of Trop-2 protein and mRNA in different tissues. The expression of Trop-2 protein in HepG2, HCCLM3 and HL-7702 cells was detected by flow cytometry and Western blot. Western blot was utilized to detect the expression of Trop-2, protein kinase C alpha(PKC-α), phosphorylated protein kinase C alpha(p-PKC-α) and nuclear factor of kappa B(NF-κB) in different tissues and transfected HepG2 cells, besides, CCK-8 was applied to determine the cell proliferation rate in transfected HepG2 cells. Result The relative expressions of p-PKC-α, NF-κB, Trop-2 protein and Trop-2 mRNA in hepatic carcinoma tissues were significantly higher than those in adjacent normal tissues(P<0.01). The relative expression levels of Trop-2 protein in HCCLM3 cells and HepG2 cells were significantly higher than those in HL-7702 cells(P<0.01). The relative expressions of Trop-2, p-PKC-α, PKC-α and NF-κB in HepG2 cells in the Trop-2 siRNA group were all lower than those in the blank control group and the negative control group(P<0.05). The proliferation rate of HepG2 cells in Trop-2 siRNA group was lower than that in the negative control group(P<0.05). Conclusion Trop-2 gene is highly expressed in hepatic carcinoma tissues and hepatic carcinoma cell lines, suggesting that it may be a potential therapeutic target for hepatic carcinoma.更多还原