Loss of mismatch repair activity in simian virus 40 large T antigen–immortalized BPH-1 human prostatic epithelial cell line
Article first published online: 20 JUL 2001
Copyright © 2001 Wiley-Liss, Inc.
Volume 31, Issue 3, pages 145–151, July 2001
How to Cite
Yeh, C.-C., Lee, C., Huang, M.-C. and Dahiya, R. (2001), Loss of mismatch repair activity in simian virus 40 large T antigen–immortalized BPH-1 human prostatic epithelial cell line. Mol. Carcinog., 31: 145–151. doi: 10.1002/mc.1049
- Issue published online: 24 JUL 2001
- Article first published online: 20 JUL 2001
- Manuscript Accepted: 3 MAY 2001
- Manuscript Revised: 5 APR 2001
- Manuscript Received: 27 NOV 2000
- mismatch repair;
- SV40 large T antigen;
Simian virus 40 large T antigen (SVLTAg) has been used to immortalize cells; however, the mechanism leading to immortalization is still unclear. We hypothesize that DNA mismatch repair (MMR) activity is important during SVLTAg-induced immortalization. To test this hypothesis, we used the SVLTAg-immortalized cell line BPH-1 derived from human benign prostate epithelial cells to analyze MMR activity and the expression of MMR genes (hMLH1, hPMS1, hPMS2, hMSH2, hMSH3, and hMSH6). The results demonstrated that BPH-1 cells were deficient in repairing G:T, A:C, and G:G mispairs in bacteriophage M13mp2. Reverse-transcription polymerase chain reaction experiments indicated MMR genes (hMSH3, hMSH6, and hPMS1) were expressed at a low level in BPH-1 cells. In contrast, all six MMR genes were expressed in human benign prostate hyperplasia tissues. Downregulation of hMSH3, hMSH6, and hPMS1 genes is not a result of the hypermethylation mechanism because demethylation with 5-aza-2′-deoxycytidine did not restore expression of these genes. Although the hMLH1 gene is expressed in BPH-1 cells, western blotting and exon analyses demonstrated that hMLH1 was mutated and/or deleted in BPH-1 cells. © 2001 Wiley-Liss, Inc.