Overexpression of lentivirus RFX1 and its inhibitory effect on proliferation of glioblastoma cells.
- Author:
Kai CHENG
1
;
Haojie SUN
2
;
Mingzhi ZHANG
2
;
Li SHEN
2
Author Information
1. Department of Biochemistry, Fenyang College of Shanxi Medical University, Fenyang Shanxi 032200; Stem Cell Research Center, Peking University Health Science Center, Beijing 100083, China.
2. Stem Cell Research Center, Peking University Health Science Center, Beijing 100083, China.
- Publication Type:Journal Article
- MeSH:
Animals;
Cell Line, Tumor;
Cell Proliferation;
genetics;
Genetic Vectors;
Glioblastoma;
genetics;
physiopathology;
HEK293 Cells;
Humans;
Lentivirus;
genetics;
Plasmids;
Rats;
Regulatory Factor X1;
genetics;
physiology;
Transfection;
Up-Regulation
- From:
Journal of Central South University(Medical Sciences)
2016;41(11):1117-1123
- CountryChina
- Language:Chinese
-
Abstract:
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.
