Reg Ⅳ gene in colorectal cancer cell lines mRNA expression and pcDNA.doc

 PAGE \* MERGEFORMAT 6 Reg Ⅳ gene in colorectal cancer cell lines mRNA expression and pcDNA Author: peak, Zheng Sheng-Qin, Zhang Wei, JIN Yueling, Tian Xiao-Qiang, Huang Pei-lin [Abstract] Objective: To explore the Reg Ⅳ gene in the four kinds of colorectal cancer cell lines the expression level of construct and transfected pcDNAReg Ⅳ recombinant plasmid, in order to clarify the Reg Ⅳ gene in colorectal cancer genesis and development of the role of laying the ground work. Methods: RTPCR to detect HT29, Caco2, Sw480, LoVo cell lines Reg Ⅳ gene expression level, will receive the Reg Ⅳ gene was cloned pcDNA3.1 / (-) plasmid, using PCR, restriction enzyme digestion were identified, the liposome Low expression of transfected into even knot straight Reg Ⅳ expression of cancer cell lines, using RTPCR identified after G418 selection. Results: HT29, Caco2 cell line Reg Ⅳ gene overexpression, Sw480 cell lines expressing weak compared with the first two, LoVo cell line Reg Ⅳ negative expression; the recombinant plasmid containing the gene Reg Ⅳ by pcDNAReg Ⅳ transfected LoVo cells showed positive expression of genes Reg Ⅳ . Conclusion: Reg Ⅳ gene in the different colorectal cancer cell lines expressed different levels. Was successfully constructed and transfected pcDNAReg Ⅳ , can make Reg Ⅳ (-) of the LoVo cell lines tested positive for Reg Ⅳ gene expression. [Keywords:] Reg Ⅳ gene; colorectal cancer; recombinant plasmid; gene expression; gene transfer Abstract: Objective To investigate the expression of Reg Ⅳ in four colorectal cancer cells and construct recombinant plasmid pcDNAReg Ⅳ and transfect it for further study of Reg Ⅳ in colorectal cancer cells.Methods Reg Ⅳ obtained from colorectal cancer cells by RTPCR was cloned into pcDNA3.1 / (-- ) to form pcDNAReg Ⅳ which was transfected into colorectal cancer cell with less or no expression of Reg Ⅳ via liposome after identification of PCR, restriction enzyme digesting and DNA sequencing. The