构建CD14shRNA慢病毒载体对胃腺癌SGC7901细胞影响.pdfVIP

构建CD14-shRNA 慢病毒载体对胃腺癌SGC7901 细胞 的影响 中 文 摘 要 目的：本研究拟构建CD14-shRNA 慢病毒载体，建立CD14 稳定低 表达的胃癌细胞系，探讨 CD14-shRNA 慢病毒载体转染胃癌 SGC-7901 细胞后CD14 的mRNA 及蛋白的表达情况。 方法：将CD14-shRNA 慢病毒载体转染胃癌SGC-7901 细胞，以最 佳感染指数（MOI ）转染胃癌 SGC-7901 细胞后，用 Real-timePCR 和 Westernblot 分别检测空白对照组、NCshRNA 阴性对照组和CD14shRNA 慢病毒载体组的CD14mRNA 和蛋白质的表达。 结果：将CD14-shRNA 慢病毒载体成功转染至胃癌细胞系，RT-PCR 结果显示转染CD14shRNA 慢病毒组CD14 的△ Ct 值比NCshRNA 对照组 增高 （p 0.000;P 0.001 ），其CD14 的2?△△ Ct 值比NCshRNA 对照组降低 p 0.0001 ；P 0.001 ，差异有显著统计学意义；Western blot 检测结果显 示，CD14shRNA 慢病毒载体组细胞的CD14 蛋白表达相对于空白组及阴 性对照组明显减少，从灰度值分析显示看出，CD14shRNA 慢病毒组CD14 蛋白相对表达量（0.01 ）远远小于空白组（1.0）及阴性对照组（0.83 ）。 结论：成功将CD14shRNA 慢病毒载体转染至胃癌SGC7901 细胞， CD14shRNA 慢病毒载体转染至胃癌 SGC790 细胞后能抑制胃癌细胞 CD14 的mRNA 及蛋白的表达。 关键词：CD14，shRNA 载体构建，胃癌细胞 The effect of the construction of CD14-shRNA lentiviral vectors on gastric cancer cells ABSTRACT OBJECTIVE: This study aimed to investigate the SGC-7901 cells, CD14 mRNA and protein level after CD14-shRNA expression vector transfected the gastric cancer cells and to and to lay a experimental basis for the study of the pathogenesis of gastric cancer. METHODS: CD14-shRNA lentiviral vector was constructed to transfect gastric cancer SGC-7901 cells. RT-PCR and Western blot was used to detect the CD14 mRNA and protein level in blank control group, negative control group and CD14shRNA lentiviral vector group. Results: CD14-shRNA expression vector was successfully constructed and transfect gastric cancer cells.RT-PCR showed that the △ Ct values of transfection CD14shRNA group was higher than NCshRNA control group P 0.000;P 0.001 , the 2?△△ Ct value of CD14shRNA group 1±0.08 was lower than NCshRNA control group p 0.0001; P 0.001 , the difference was statistically significant. Western blot analysis showed that CD14shRNA lentiviral group relative expression of CD14 protein was 0.01, that far less than the NCshRNA negative