Construction and application in renal cell carcinoma 769-P cells of miR-218 expression plasmids

VernacularTitle:miR-218表达质粒的构建及其在肾癌769-P细胞中的表达
Author: Long HE ; Yan ZHANG ; Jun WANG ; Long LIU ; Lianhui FAN
Publication Type:Journal Article
Keywords: miR-218; Plasmids; Carcinoma,renal cell
From: Cancer Research and Clinic 2013;25(12):806-808,811
CountryChina
Language:Chinese
Abstract: Objective To construct a stable expression plasmids miR-218,to lay the experimental foundation for the expression and mechanism of miR-218 in human renal cell carcinoma.Methods The primers were designed by Has-miR-218 and the miR-218 fragment were amplified by PCR,and then which was connected with the carrier pcDNA3.1 (+) to build a stable expression plasmids.The recombinant plasmids were trasfected into renal cell carcinoma cell line 769-P,and the expression of miR-218 in these cells were detected.Results The plasmids were constructat successfully,which was determind by enzyme digestion and sequencing results.The constructed expression plasmids pcDNA3.1-miR-218 transfection of human renal cell carcinoma cell line 769-P,miR-218 cxpression level of cells was significantly increased after trasfected by recombinant plasmids carrying miR-218 gene (relative expression quantity was 1.64).Conclusion The miR-218 expression plasmids pcDNA3.1-miR-218 were successfully constructed,the plasmids can be applied to the study of miR-218 function and mechanism in renal cell carcinoma.