人突变K-ras基因修饰lewis肺癌细胞株的建立及其生物学特性

目的:探讨以K-ras基因为靶点,建立人K-ras(12位密码子点突变)基因修饰的lewis肺癌细胞株3LL-pcDNA3-K-ras/V12及其生物学特性.方法:应用脂质体法将人K-ras(12位密码子点突变)基因cDNA导入lewis肺癌细胞株3LL,应用流式细胞仪检测p21/V12蛋白的表达,同时检测3LL细胞株转基因前后生长、克隆形成率和体内成瘤的变化.结果:成功的将人K-ras(12位密码子点突变)基因cDNA导入lewis肺癌细胞株3LL,p21/V12蛋白在3LL-pcDNA3-K-ras/V12中稳定表达,转基因细胞株的生长曲线和克隆形成率与原代细胞株3LL无显著差异;但转基因细胞株在C57BL/6小鼠体内的成瘤性显著高于原代细胞株3LL.结论:成功的建立了导入K-ras(12位密码子点突变)的lewis肺癌细胞株3LL-pcDNA3-K-ras/V12.

Establishment and Biological characteristics of Lewis Lung Cancer Cell Line Modified by Mutant K-ras Gene

Objective: To establish a Lewis lung cancer cell line modified by human mutant K-ras gene and investigate the biological characteristics of this cell line. Methods: K-ras/V12 gene was transfected into cultured Lewis lung cancer cell line by the lipofectin-mediated gene transfection method. The gene expression and proliferation characteristic of the transgenic cell were observed in vitro, and the ability of tumorigenesis was observed in vivo. Results: K-ras/V12 gene was successfully transfected into cultured Lewis lung cancer cell line by the lipofectin-mediated gene transfectionmethod. The transgenic cell had no significant difference with Lewis lung cancer cell line in grow curve and soft agar colony forming rate. This cell had higher expression level of p21 and had higher ability of tumorigenesis in C57BL/6 mice. Conclusion: The K-ras/V12 modified Lewis lung cancer cell line is successfully established.