摘要
目的:观察CD14-shRNA慢病毒载体转染SGC-7901细胞后CD14 mRNA及蛋白变化,为胃癌发病机制的研究奠定实验基础.方法:将构建好的CD14-shRNA慢病毒载体,转染SGC-7901细胞后,用Real-time PCR和Western blot分别检测空白对照组、NC-CD14shRNA阴性对照组及CD14-shRNA慢病毒载体组的CD14 mRNA和蛋白质的表达.结果: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CD14-shRNA慢病毒组CD14的2Ct值(1±0.08)比NCshRNA对照组2C t值(0.16±0.02)降低(P=0.0001;P<0.001),差异有统计学意义;Western blot检测结果显示,CD14-shRNA慢病毒组CD14蛋白相对表达量为0.01,远远小于SGC-7901空白组(1.00)及NCshRNA阴性对照组(0.83).结论:CD14-shRNA慢病毒载体转染至胃癌SGC-7901细胞后能抑制胃癌细胞CD14 mRNA及蛋白表达,CD14在胃癌发生、发展中具有重要作用,有望为胃癌防治提供新的思路.
AIM： To develop a SGC-7901 cell line with stably silenced expression of CD14 to provide an ex- perimental basis for the study of the pathogen- esis of gastric cancer. METHODS： A CD14-shRNA lentiviral vec- tor was constructed to transfect gastric cancer SGC-7901 cells. RT-PCR and Western blot were used to detect the CD14 mRNA and protein levels in untreated SGC-7901 cells （blank con- trol group）, SGC-7901 cells transfected with NC-CD14 shRNA （NC-CD14 shRNA control group） and SGC-7901 cells transfected with the CD14-shRNA lentiviral vector （CD14-shRNA lentiviral vector group）.RESULTS： CD14-shRNA expression vector was successfully constructed and transfected into gastric cancer cells. RT-PCR showed that the Ct value for the CD14-shRNA lentiviral vector group was significantly higher than those for the NC-CD14 shRNA control group and blank con- trol group （P = 0.000; P 〈 0.001）, and the 2Ct value for the CD14-shRNA lentiviral vector group （1± 0.08） was significantly lower than those for the NC-CD14 shRNA control group and blank con- trol group （P = 0.0001; P 〈 0.001）. Western blot analysis showed that the relative expression of CD14 protein in the CD14-shRNA lentiviral vec- tor group （0.01） was significantly less than that in the blank control group （1.0） and NC-CD14 shRNA control group （0.83）. CONCLUSION： A CD14-shRNA lentiviral vec- tor has been successfully transfected into gas- tric cancer SGC-7901 cells, which can inhibit the expression of CD14 mRNA and protein in SGC-7901 ceils.
出处
《世界华人消化杂志》
CAS
北大核心
2014年第22期3194-3200,共7页
World Chinese Journal of Digestology
基金
国家自然基金资助项目
No.81360531
广西自然基金资助项目
No.2010GXNSFA013219
广西卫生厅重点基金资助项目
No.重2012030~~
