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PubMed | Key laboratory of Molecular Tumor Pathology of Guangdong Province
Type: Journal Article | Journal: Nan fang yi ke da xue xue bao = Journal of Southern Medical University | Year: 2015

To construct a recombinant lentivirus vector for Wilms tumor on X chromosome (WTX) gene and establish a colorectal cancer SW620 cell line with stable WTX over-expression.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.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.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.


Ma W.,Key laboratory of Molecular Tumor Pathology of Guangdong Province | He L.,Key laboratory of Molecular Tumor Pathology of Guangdong Province | Liu C.,Key laboratory of Molecular Tumor Pathology of Guangdong Province | Zhang Q.,Key laboratory of Molecular Tumor Pathology of Guangdong Province | Ding Y.,Key laboratory of Molecular Tumor Pathology of Guangdong Province
Nan fang yi ke da xue xue bao = Journal of Southern Medical University | Year: 2015

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.

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