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目的研究功能未知基因DIAPH3在肝细胞癌(肝癌)组织中的表达及其与肝癌细胞生长的关系。方法利用肝癌基因表达谱数据研究发现DIAPH3基因在肝癌组织中高表达。采用RT-PCR及实时荧光定量PCR(RFQ-PCR)检测DIAPH3在人类正常组织以及肝癌组织和癌旁组织中的表达。RNA干扰技术沉默肝癌细胞株(7701和7402)内DIAPH3基因的表达,细胞计数试剂盒8(CCK-8)实验定量分析内源性DIAPH3基因被沉默前后的细胞生长曲线的变化。结果 DIAPH3基因在75%(18/24)的肝癌组织中有明显的上调。沉默肝癌细胞株7701和7402中内源性DIAPH3基因表达后,两种肝癌细胞的生长均受到明显的抑制(P<0.01)。结论 DIAPH3可能是一个新的肿瘤相关癌基因,在维持肝癌细胞恶性表型过程中可能起到重要的作用。
Objective To study the expression of DIAPH3 gene in hepatocellular carcinoma (HCC) and its relationship with the growth of HCC cells. Methods Using the gene expression profiling data of liver cancer, DIAPH3 gene was found to be highly expressed in HCC tissues. The expression of DIAPH3 in human normal tissues, hepatocellular carcinoma tissues and adjacent tissues was detected by RT-PCR and real-time quantitative PCR (RFQ-PCR). The RNA interference technique was used to silence the expression of DIAPH3 gene in hepatocellular carcinoma cell lines (7701 and 7402). The cell counting kit 8 (CCK-8) assay was used to quantitatively analyze the changes of cell growth curve before and after the silencing of endogenous DIAPH3 gene. Results DIAPH3 gene was significantly up-regulated in 75% (18/24) of HCC tissues. The silencing of endogenous DIAPH3 gene expression in silencing hepatoma cell lines 7701 and 7402 both inhibited the growth of both hepatocellular carcinoma cells (P <0.01). Conclusion DIAPH3 may be a new tumor-associated oncogene, which may play an important role in maintaining the malignant phenotype of hepatocellular carcinoma cells.