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

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pITA，构建pITA-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pITA-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。

Overexpression of lentivirus RFX1 and its inhibitory effect on proliferation of glioblastoma cells

Objective: To construct overexpression lentivirus vector for human regulatory factor X1 (RFX1) gene, and to explore its effect on proliferation of F98 cell line.Methods: Huamn RFX1 gene was amplified by polymerase reaction. Gene amplification products were inserted into lentivirus vector pITA, and the lentivirus vector pITA-RFX1 was constructed. The constructed vector was verified by agarose gel electrophoresis and DNA sequencing. Lentivirus vector pITA-RFX1 and virus packaging plasmids were cotransfected into 293T cells, and then transfected into F98 cells. RFX1 protein expression were detected by Western blot and laser confocal before and after transfection. Flow cytometry and cell counting kit-8 were used to detect cellular proliferation.Results: Agarose gel electrophoresis and DNA sequencing showed that recombinant lentivirus plasmids pITA-RFX1 were constructed successfully. After transfection of pITA-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.