Growth suppression of human gastric carcinoma cell line SGC-7901 by stable transfection of antisense angiopoietin2 gene
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Growth suppression of human gastric carcinoma cell line SGC-7901 by stable transfection of antisense angiopoietin2 gene
Journal of Chongqing Medical UniversityVol. 36, Issue 5, Pages: 497-501(2013)
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重庆医科大学附属第二医院普外科,重庆,400010
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Published:2013,
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CHEN Lei, ZHU Peng. Growth suppression of human gastric carcinoma cell line SGC-7901 by stable transfection of antisense angiopoietin2 gene. [J]. Journal of Chongqing Medical University 36(5):497-501(2013)
DOI:
CHEN Lei, ZHU Peng. Growth suppression of human gastric carcinoma cell line SGC-7901 by stable transfection of antisense angiopoietin2 gene. [J]. Journal of Chongqing Medical University 36(5):497-501(2013)DOI:
Growth suppression of human gastric carcinoma cell line SGC-7901 by stable transfection of antisense angiopoietin2 gene
Objective:To evaluate expressions of angiopoietin2(Ang2) protein in human gastric cancer cells SGC-7901 transfected by pEPFP-N1-antisense Ang2 and to investigate whether antisense Ang2 gene could inhibit the vitro growth of human gastric carcinoma cell line. Methods:Two different plasmids including pEGFP-N1 and pEGFP-N1 SGC-7901 antisense Ang2 were separately trans-ferred into an in vitro cultured human gastric cancer cell line SGC-7901 using Lipofectamine2000 transfection technique. After trans-fection,the cells were selected by G418. Then resistant clones were chosen and expanded in DMEM culture medium,with parental SGC-7901 cells as control. RT-PCR and immunohistochemical method were done to determine whether target genes and its proteins had expressed. Cell viability was determined by MTT assay. Apoptosis was determined by flow cytometry with a double dyeing method using FITC-conjugated annexin V and PI. Results:According to results of RT-PCR and immunohistochemical methods,angiopoietin mRNA and its proteins were not expressed in transfected antisense Ang2 gene SGC-7901 cells. MTT assay showed that pEGFP-N1-anti Ang2 transfection group had much lower cell viability than other groups(F=10.39,P=0.002 4). Meanwhile,pEGFP-N1-anti Ang2 transfection group had an increase in apoptosis rates compared with those of the other groups based on results of flow cytometry(?字2=3 188.98,P<0.000 1). Conclusions:pEGFP-N1-anti Ang2 successfully transfects human gastric cancer cells SGC-7901,which in-hibits Ang2 mRNA and protein expressions as well as suppresses the malignant phenotype of human gastric cells.