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TOP2A在肝癌中的表达及其与预后的关系研究
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Abstract:
目的：探讨DNA拓扑异构酶IIα (topoisomerase II alpha, TOP2A)在肝癌中的表达及其对预后的影响。方法：利用Oncomine数据库分析TOP2A基因在肝癌和正常肝组织中的差异表达。用Western blot和qRT-PCR检测TOP2A在收集的肝癌标本和肝癌细胞系中的表达水平。此外，我们还建立了稳定的肝癌细胞系，在肝癌细胞中下调或过表达TOP2A。运用CCK-8法检测该基因表达如何影响细胞增殖能力。借助GEPIA网站对肝癌数据集进行与TOP2A相关的生存分析。结果：肝癌组织或细胞系中TOP2A的表达水平显著高于正常肝组织及细胞；敲除TOP2A表达可降低原本高表达该基因的肝癌细胞的增殖活力，而上调其表达对原本低表达该基因的细胞的增殖有促进作用，且TOP2A表达量高的患者，其无病生存期(DFS)及总生存期(OS)均较短。结论：TOP2A可能参与肝癌的发生发展，可作为新型肝癌预后标志物，为肝癌精准治疗策略选择提供重要依据。
Objective: To investigate the expression of DNA topoisomerase II alpha (TOP2A) in hepatocellular carcinoma (HCC) and its effect on prognosis. Methods: The differential expression of TOP2A in hepatocellular carcinoma and normal liver tissues was analyzed by Oncomine database. Then, we used western blot and qRT-PCR to detect the expression level of TOP2A in collected liver cancer samples and liver cancer cell lines. In addition, we also established a set of stable hepatocellular carcinoma cell lines, which down-regulated or overexpressed TOP2A in them. CCK-8 test was operated to detect how the expression of the gene affected the ability of cell proliferation. The survival analysis of liver cancer data set related to TOP2A was carried out with the assistance of GEPIA. Results: Four studies on the expression of TOP2A in HCC tissues and normal tissues (Roessler Liver, Chen Liver, Wurmbach Liver, Roessler Liver 2) were screened from Oncomine database. The expression level of TOP2A in liver cancer group was higher than that in normal tissue group in 4 cohorts. In a cohort of 40 liver cancer and paracancerous tissue samples we collected after radical resection, the expression of TOP2A mRNA was up-regulated in 33 tumor samples and down-regulated in 7 tumor samples. In addition, we also detected the expression of TOP2A in human normal liver cell line and hepatocellular carcinoma cell line. The results showed that the mRNA and protein levels of TOP2A in high metastatic cell lines MHCC97H, HuH7 and LM3 were significantly higher than those in low metastatic cell line Hep3B and normal liver cell line L02. We chose MHCC97H with relatively high expression of TOP2A and Hep3B with relatively low expression of TOP2A for follow-up experiments. To assess the effect of TOP2A on the viability of MHCC97H and Hep3B cells, the absorbance values (OD) of MHCC97H and Hep3B at 450 nm were recorded by CCK8 assay after transfection by the shTOP2A and pcDNA-TOP2A respectively, and the results showed that TOP2A knockout could reduce the viability of MHCC97H cells, while up-regulation of TOP2A expression promoted the proliferation of Hep3B cells (p < 0.01).
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