肿瘤相关抗原编码基因CHP2对细胞增殖的抑制作用

Tumor Associated Antigen Encoding Gene CHP2 Inhibited Cell Proliferation

目的:研究肿瘤相关抗原编码基因CHP2对细胞增殖作用的影响.方法:采用PCR方法获得目的基因,克隆至真核表达载体pcDNA3,对产生的重组子进行酶切鉴定及DNA序列测定;测序正确的重组子用脂质体转染的方法转入293细胞系,药物G418筛选稳定转染细胞株,分别在基因水平以及蛋白水平进行鉴定;用3H-TdR掺入的方法检测CHP2对细胞增殖的影响.结果:经琼脂糖凝胶电泳及测序鉴定,所插入的片断方向及序列完全正确;Western blotting结果显示,获得的稳定转染细胞株能够高效的表达CHP2蛋白.细胞增殖实验结果显示CHP2稳定转染细胞株的3H-TdR掺入值明显低于对照组.结论:CHP2在细胞中的有效表达能够显著显著抑制细胞的增殖,CHP2可能是参与细胞生长调控的一个重要的调节因子.

Objective: To study the effect of cell proliferation transfected with tumor associated antigen encoding gene CHP2. Methods: Using PCR method to clone CHP2 encoding gene and inserting it into the eukaryotic expression vector pcDNA3 to construct the recombination pcDNA-CHP2. The fusion gene was identified by enzyme digestion and DNA se- quencing. 293 cells transfected CHP2 genes were selected by G418 pressure and the cell proliferation was measured by ~3- TdR uptake. Results: Identified by enzyme digestion and sequencing, CHP2 gene was correctly eloned into pcDNA3. The results of the western blot showed the transfected cell could express CHP2 protein. ~3-TdR uptake value of the CHP2 trans- fected cell was lower than the control. Conclusions: CHP2 could inhibit cell proliferation and it maybe an important regu- lator involved in cell proliferation.