肝癌组织CT抗原基因CSAG1表达及体外对肝癌细胞生长的影响

Expression of CT antigen gene CSAG1 in hepatocellular carcinoma tissues and its effect on growth of hepatocellular carcinoma cell in vitro

目的:探讨CSAG1(chondrosar-coma associated gene 1)基因在肝癌组织中的表达及其对肝癌细胞生长增殖的影响,为了解肝癌发生的分子机制奠定理论基础。方法:利用半定量RT-PCR技术检测CSAG1基因在12例肝癌和癌旁组织中的表达差异;RNA干扰技术沉默肝癌细胞株(Focus)内CSAG1基因的表达;利用CCK-8、克隆形成实验以及流式细胞仪分别定量分析内源性CSAG1基因被沉默前后的Focus细胞生长曲线、克隆形成能力以及细胞周期的变化。结果:与癌旁组织相比较,CSAG1基因在75%(9/12)肝癌组织中的表达量明显上调;沉默肝癌细胞株Focus中内源性CSAG1基因表达后,Focus细胞的生长明显减慢,克隆形成能力明显降低,S期细胞明显减少。结论:CSAG1可能是一个新的肝癌相关的促进基因,对其进行深入的研究有可能发现肝癌发病的新机制,也有可能发现肝癌治疗的新靶点。

OBJECTIVE： To study the expression of CSAG1 gene in hepatocellular carcinoma （HCC） tissues and the silencing effect of CSAG1 gene on the cell growth of focus, and a cell line derived from liver cancer. METHODS： The expression of CSAG1 was investigated through semi quantitative RT-PCR in 12 pairs of HCCs compared with adjacent non-cancerous livers （non HCC）. The expression of CSAG1 gene was valuated in 6 HCC derived cell lines, one fetal liver tissue and one normal liver tissue by RT-PCR, respectively. The cell growth and colony formation were observed as the silencing of CSAG1 by a shRNA method. Simultaneously, the cell cy ele was detected through the fluorescence activated cell sorter （FACS）. RESULTS： CSAG1 was significantly up-regulated in 75 % （9/12） HCC specimens at the mRNA level compared with non- HCCs. The silencing of CSAG1 gene in focus cells markedly inhibited the cell growth, weakened the ability of colony formation, and significantly decreased S phase population compared with the control cells with the endogenous CSAG1 expression. CONCLUSIONS： The up regulation of CSAG1 as a tumor-associated gene can contribute to the oncogenesis of HCC. It may be an important event in hepatocarcinogensis and may be a new candidate target in HCC treatment.