南方医科大学学报2015，Vol.35Issue(8)：1122-1127,6.
应用重组慢病毒表达载体建立抑癌基因WTX稳定高表达结直肠癌SW620细胞系
Establishment of a colorectal cancer SW620 cell line stably over-expressing Wilm's tumor on X chromosome using a recombinant lentivirus vector
摘要
Abstract
Objective To construct a recombinant lentivirus vector for Wilm's tumor on X chromosome (WTX) gene and establish a colorectal cancer SW620 cell line with stable WTX over-expression. Methods The full length coding region of WTX gene was amplified with PCR, and the amplified fragment was cloned into the lentivirus vector GV387. The recombinant lentivirus vector was transfected in 293T cells for packaging the virus, which was then transfected into colorectal cancer SW620 cells. The stably transfected cells were selected with G418, and the cellular expressions of WTX mRNA and protein were detected using quantitative PCR and Western blotting. Results The recombinant plasmid was successfully constructed as verified by sequence analysis. Quantitative PCR and Western blotting results showed that trasnfection with the recombinant lentivirus significantly increased the expression levels of WTX in SW620 cells. Conclusion We successfully established a colorectal cancer cell lines with stable over-expression of WTX, which provides an essential cell model for studying the role of WTX in the tumorigenesis and progression of colorectal cancer.关键词
Wilm'stumoronXchromosome/慢病毒载体/稳定转染/结直肠癌Key words
Wilms tumor on X chromosome/lentivirus vector/transfection/colorectal cancer引用本文复制引用
马文霞,贺莉,刘椿,张庆玲,丁彦青..应用重组慢病毒表达载体建立抑癌基因WTX稳定高表达结直肠癌SW620细胞系[J].南方医科大学学报,2015,35(8):1122-1127,6.基金项目
国家自然基金(81472712,81071989) （81472712,81071989）
广东省科技计划项目(c1221020700008) （c1221020700008）
山西省自然科学基金(2014011037-5) Supported by National Natural Science Foundation of China (81472712, 81071989). （2014011037-5）
