摘要
目的构建pLVX-Puro-CCR7稳定表达系统并研究CCR7基因在胃癌转移中的作用。方法采用PCR方法扩增CCR7基因并构建pLVX-Puro-CCR7重组质粒,通过脂质体转染法瞬时转染SGC7901细胞株,嘌呤霉素对该细胞株进行筛选并建立稳定表达细胞株。Western blot方法检测稳定转染的SGC7901细胞中CCR7表达情况。Transwell方法分析过度表达CCR7基因对SGC7901胃癌细胞体外迁移的影响。活体成像观察过度表达CCR7的SGC7901胃癌细胞株在裸鼠体内转移能力。结果成功构建p LVX-Puro-CCR7重组质粒并稳定转染SGC7901细胞株,该细胞株能够正确翻译及表达CCR7蛋白。过度表达CCR7基因的SGC7901胃癌细胞株体外迁移能力和在裸鼠体内转移能力显著增强(P<0.05)。结论 CCR7可能参与促进胃癌细胞转移的功能。
Objective To construct stable expression system of pLVX-Pro-CCR7 and study the function of CCR7 gene in metastasis of stomach cancer. Methods PCR method was used to amplify CCR7 gene and construct pLVX-Puro- CCR7 recombinant plasmid. Lipofection was used to infect SGC7901 cell strain immediately, and puromycin was used to screen the cell strain and establish a cell strain with stable expression. Western blot method was used to test the ex- pression of CCR7 in SGC7901 cells which were stably infected. Transwell method was used to analyze the effect of overly expressed CCR7 gene on in vitro migration of stomach cancer cells of SGC7901. Living imaging was used to ob- serve the metastasis ability of stomach cancer cell strain of SGC7901 which overly expressed CCR7 in nude mice. Re- sults pLVX-Puro-CCR7 recombinant plasmid was successfully constructed and SGC791 cell strain was stably infected. The cell strain was able to correctly translate and express CCR7 protein. Metastasis ability in nude mice and in vitro migration ability of SGC7901 stomach cancer cell strain which overly expressed CCR7 gene significantly improved （P〈 0.05）. Conclusion CCR7 may be involved in the function of promoting metastasis of stomach cancer cells.
出处
《中国现代医生》
2015年第15期14-16,20,F0003,共5页
China Modern Doctor
基金
国家自然青年基金(81301406)
浙江省自然青年基金(LQ13H190005)
