【摘要】 目的:研究锌指蛋白281(zinc finger protein 281,ZNF281)对肝癌细胞增殖的作用。方法:用realtime PCR检测80例健康人群及206例肝癌患者外周血单个核细胞中ZNF281的mRNA表达水平,并分析肝癌患者单个核细胞中ZNF281的表达水平与多个临床病理参数的相关性。采用real-time PCR及Western blot实验分别检测ZNF281在肝癌细胞和永生化肝细胞中的mRNA及蛋白表达水平。利用小干扰RNA靶向沉默Huh-7细胞和SKHep-1细胞中ZNF281的表达,然后用MTS法检测肝癌细胞活力的变化;EdU标记实验检测肝癌细胞增殖能力的变化;平板集落形成实验检测肝癌细胞集落形成能力的变化;软琼脂集落形成实验检测肝癌细胞锚定非依赖生长能力的变化。结果:与正常健康人群相比,肝癌患者的外周血单个核细胞中ZNF281的mRNA表达水平明显增高,且其增高水平与肿瘤的大小、分期以及血管侵袭性存在正相关。肝癌细胞中ZNF281表达水平高于永生化肝细胞(P<0.05)。沉默ZNF281可以抑制肝癌细胞的活力,抑制肝癌细胞增殖,DNA合成降低,集落数量减少,锚定非依赖性生长能力减弱(P<0.05)。结论:ZNF281可促进肝癌细胞的增殖。更多还原

【Abstract】 AIM: To investigate the role of zinc finger protein 281（ ZNF281） in the proliferation of hepatocellular carcinoma（ HCC） cells. METHODS: The mRNA expression levels of ZNF281 in peripheral blood mononuclear cells from 80 cases of healthy people and 206 cases of HCC patients were determined by real-time PCR. Statistical analysis were used to illustrate the relationship between the mRNA expression levels of ZNF281 in the peripheral blood mononuclear cells and the clinicopathologic parameters of HCC patients. Real-time PCR and Western blot were used to detect the mRNA and protein expression levels of ZNF281 in hepatoma cell lines and immortalized hepatocytes. The silencing of ZNF281 was conducted by transfection of small interfering RNA targeting ZNF281,and then the proliferation of HCC cells was analyzed by MTS assay. The DNA synthesis of HCC cells was tested by Cell-Light^TM Ed U Apollo^? 488 In Vitro Imaging Kit. The ability of colony formation of the HCC cells was measured by colony formation assay,and the ability of anchorage-independent growth was detected by soft agar test. RESULTS: The mRNA expression level of ZNF281 in the peripheral blood mononuclear cells from HCC patients was significantly increased compared with the healthy people,and the high expression level was positively correlated with tumor size,tumor stage and tumor vascular invasion. Concurrently,the expression level of ZNF281 in hepatoma cell lines was significantly higher than that in immortalized hepatocytes. More importantly,the silencing of ZNF281 inhibited the proliferation,DNA synthesis,colony formation and anchorage-independent growth of the HCC cells. CONCLUSION: ZNF281 promotes the proliferation of HCC cells.更多还原