Carcinogenesis, Teratogenesis & Mutagenesis ›› 2001, Vol. 13 ›› Issue (2): 92-095.doi: 10.3969/j.issn.1004-616x.2001.02.008

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ESTABLISHMENT OF BCL-2 STABLY-EXPRESSED HEPATOMA CELL STRAIN BY GENE TRANSFECTION METHOD

YANG Lian-jun, WANG Wen-liang   

  1. Department of Pathology, Fourth Military Medical University, Xi'an 710032, China
  • Received:2000-07-11 Revised:2000-11-22 Online:2001-04-30 Published:2001-04-30

Abstract: Purpose and Methods : To establish a hepatocellular carcinoma (HCC) cell st rain that expresses bcl2-protein stably the ret rovirus expression vector pDOR2SB containing human bcl-2 cDNA was t ransduced into bcl-2 protein negative cell line HCC-9204 by lipid2mediated gene t ransfection method. The bcl-2 t ransfected cells were obtained after being selected with G-418 , and the expression of bcl-2 was detected by immunocytochemist ry after the cells being expanded. The t ransfected cells were cloned by a limited dilution method , and bcl-2 expression in the monoclonal cells was detected with flow cytometer. Results : Bcl-2 protein was expressed in most of the HCC-9204 cells t ransfected with pDOR2SB , but was negative in the HCC-9204 cells t ransfected with pDOR vector and nont ransfected HCC29204 cells. After being cloned for 3 times continually , the monoclonal cell strain that expressed bcl-2 protein at a 100 % positive rate was obtained. Conclusion : The HCC cell st rain that expressed bcl-2 protein stably , was established successfully by gene t ransfection method.

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