慢病毒介导的调节因子XⅠ过表达及其对胶质瘤细胞增殖的抑制作用  

Overexpression of lentivirus RFXⅠ and its inhibitory effect on proliferation of glioblastoma cells

作　　者：程凯[1,2] 孙浩杰[2] 张明智[2] 沈丽[2] 

机构地区：[1]山西医科大学汾阳学院生物化学教研室,山西汾阳032200 [2]北京大学医学部干细胞中心,北京100083

出　　处：《中南大学学报（医学版）》2016年第11期1117-1123,共7页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(81272790);山西医科大学汾阳学院科技发展基金(2016B05)~~

摘　　要：目的:构建调节因子X1(regulatory factor X1,RFX1)过表达慢病毒载体,并观察其对F98细胞增殖的影响。方法:运用PCR技术扩增RFX1,并将其插入慢病毒空载体质粒p ITA,构建p ITA-RFX1慢病毒质粒,经琼脂糖凝胶电泳和测序鉴定正确。将包装慢病毒共转染293T细胞,然后感染F98细胞。利用Western印迹和激光共聚焦验证RFX1过表达情况,用流式细胞仪和CCK-8试剂盒观察转染前后细胞增殖情况。结果:电泳和测序显示慢病毒载体p ITA-RFX1构建成功,将其感染F98细胞后过表达RFX1蛋白,同时可以明显抑制细胞的增殖。结论:过表达慢病毒载体构建成功,上调RFX1可以降低恶性胶质瘤细胞的增殖。Objective： To construct overexpression lentivirus vector for human regulatory factor X1（RFX1） gene, and to explore its eff ect on proliferation of F98 cell line.Methods： Huamn RFX1 gene was amplified by polymerase reaction. Gene amplification products were inserted into lentivirus vector pI TA, and the lentivirus vector pI TA-RFX1 was constructed. Th e constructed vector was verified by agarose gel electrophoresis and DNA sequencing. Lentivirus vector p ITA-RFX1 and virus packaging plasmids were cotransfected into 293 T cells, and then transfected into F98 cells. RFX1 protein expression were detected by Western blot and laser confocal before and af ter transfection. Flow cytometry and cell counting kit-8 were used to detectcellular proliferation.Results： Agarose gel electrophoresis and DNA sequencing showed that recombinant lentivirus plasmids pI TA-RFX1 were constructed successfully. After transfection of pI TA-RFX1, the RFX1 protein were over-expressed, which significantly inhibited the proliferation of F98 cells.Conclusion： The overexpression lentivirus vector for RFX1 was constrcted successfully, and the up-regulation of RFX1 can prevent the proliferation of glioblastoma cells.

关 键 词：调节因子X1 慢病毒 过表达 恶性胶质瘤 

分 类 号：R739.41[医药卫生—肿瘤]