肿瘤抗原基因OVA66对肿瘤细胞生物学特征的影响

目的:探讨肿瘤抗原基因OVA66对肿瘤细胞生物学特征的影响。方法:采用脂质体法,以重组真核表达载体pcDNA3.1-OVA66转染肝癌细胞系SMMC-7721,经G418稳定筛选、克隆及扩增,采用RT-PCR、Westernblot和免疫细胞化学(ICC)染色法,检测目的基因及其蛋白的表达。用FACS检测细胞周期的变化;电镜技术观察细胞的超微结构;体外运动、侵袭实验检测细胞运动及侵袭能力;并应用基因芯片技术检测相关基因的差异表达。结果:RT-PCR检测显示,转染pcDNA3.1-OVA66的细胞中OVA66基因呈高表达。Westernblot显示,在OVA66蛋白处出现明显的条带。电镜和FACS检测表明,该基因可增强肿瘤细胞的增殖能力。体外运动、侵袭实验证实,该基因可促进肿瘤细胞的迁移、侵袭。基因芯片显示,该基因的高表达可促进Ⅰ型纤溶酶原激活物抑制剂(PAI-1)等基因的表达,影响肿瘤的侵袭和转移。结论:OVA66基因及其蛋白可促进肿瘤细胞的增殖、侵袭、转移等一系列生物学功能。

The effects of tumor antigen gene OVA66 on biological characteristics of tumor cells

AIM: To explore the effects of tumor antigen gene OVA66 on the biological characteristics of tumor cells. METHODS: The eukaryotic expression vector pcDNA3.1-mock and recombined vector pcDNA3.1-OVA66 were transfected into hepatoma cell line SMMC-7721 by LipofectAMINETM 2000. After stable selection and clone proliferation of cells, the expression of OVA66 gene and protein was detected by RT-PCR, Western blot and immunocytochemical (ICC) staining. The cell growth cycle was analyzed by FACS. The ultrastructure was observed under electron microscope. The experiments in cell migration and invasion were performed in vitro. The OVA66-associated genes were analyzed by genechip technique after gene transfection. RESULTS: OVA66 was highly expressed in 7721 cells transfected with pcDNA3.1-OVA66 at mRNA level. Western blot result demonstrated that OVA66 protein content was notably increased compared with that in control cells. Electron microscope and FACS results indicated that OVA66 gene promoted the growth and proliferation of tumor cells. In vitro, the experiments in cell migration and invasion showed that this gene enhanced migration and invasion of tumor cells. The result of genechip demonstrated that the over- expression of OVA66 gene increased the expression of plasminogen activator inhibitor-1 gene (PAI-1), suggesting that it had some effects on tumor invasion and metastasis. CONCLUSION: Tumor antigen gene OVA66 and it's protein demonstrate a series of biological characteristics of tumor cells, which can promote the proliferation, migration and invasion of tumor cells.