Inhibitory effect of endostatin expressed by human liver carcinoma SMMC7721 on endothelial cell proliferation in vitro

World Journal of Gastroenterology : WJG
Xuan WangGen-Xin Xu

Abstract

To construct a stable transfectant of human liver carcinoma cell line SMMC7721 that could secret human endostatin and to explore the effect of human endostatin expressed by the transfectant on endothelial cell proliferation. Recombinant retroviral plasmid pLncx-Endo containing the cDNA for human endostatin gene together with rat albumin signal peptide was engineered and transferred into SMMC7721 cell by lipofectamine. After selection with G418, endostatin-transfected SMMC7721 cells were chosen and expanded. Immunohistochemical staining and Western blot were used to detect the expression of human endostatin in transfected SMMC7721 cells and its medium. The conditioned medium of endostatin-transfected and control SMMC7721 cells were collected to cultivate with human umbilical vein endothelial cells for 72 hours. The inhibitory effect of endostatin, expressed by transfected SMMC7721 cells, on endothelial proliferation in vitro was observed by using MTT assay. A 550 bp specific fragment of endostatin gene was detected from the PCR product of endostatin-transfected SMMC7721 cells. Immunohistochemistry and Western blot analysis confirmed the expression and secretion of foreign human endostatin protein by endostatin-transfected SMMC77...Continue Reading

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Citations

Feb 5, 2004·World Journal of Gastroenterology : WJG·Jia-Ren LiuYou-Qiang Song
Sep 13, 2003·World Journal of Gastroenterology : WJG·Yun-Feng PiaoPu-Jun Gao
Jan 13, 2004·World Journal of Gastroenterology : WJG·Jia-Ren LiuYou-Qiang Song
Apr 30, 2003·World Journal of Gastroenterology : WJG·Qin SuChun-Mei Wang
Apr 30, 2003·World Journal of Gastroenterology : WJG·Li-Wu LinXiao-Dong Lin
Jan 1, 2003·World Journal of Gastroenterology : WJG·Nan-Hong TangXiao-Zhong Wang

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