论文部分内容阅读
目的利用高通量双萤光素酶报告基因系统筛选与肿瘤转移相关的人类功能基因。方法利用与肿瘤转移相关的重要因子——血管内皮生长因子(VEGF),将含有VEGF反应序列的萤光素酶报告基因(VEGF-LUC)质粒与409个待筛人类功能基因真核表达质粒共转染Hela细胞以筛选影响VEGF-LUC表达的人类功能基因,进一步通过Western blot在蛋白质水平验证初筛阳性的基因对细胞内VEGF蛋白表达量的影响。结果染色质重塑蛋白4A(CHMP4A)明显增强VEGF-LUC相对萤光素酶值(P<0.05),Western blot结果表明CHMP4A明显增强Hela细胞内VEGF的表达。结论初步筛选到一个可能通过影响VEGF活性增强肿瘤侵袭性的基因CHMP4A。
Objective To screen human functional genes related to tumor metastasis by using high-throughput luciferase reporter gene system. Methods VEGF-LUC plasmid containing the VEGF response sequence and 409 eukaryotic expression plasmids of human functional gene to be screened were co-cultured with vascular endothelial growth factor (VEGF), an important factor related to tumor metastasis. Hela cells were transfected to screen human functional genes that affect the expression of VEGF-LUC. Western blot was used to verify the effect of primary screening positive genes on the expression of VEGF in the cells. Results Chromatin remodeling protein 4A (CHMP4A) significantly increased the relative luciferase activity of VEGF-LUC (P <0.05). Western blot results showed that CHMP4A significantly enhanced the expression of VEGF in Hela cells. Conclusion Preliminary screening of a gene CHMP4A that may enhance tumor invasiveness by affecting VEGF activity.