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World J Gastroenterol. Dec 7, 2007; 13(45): 6053-6059
Published online Dec 7, 2007. doi: 10.3748/wjg.v13.i45.6053
Effect of Bcl-2 and Bax on survival of side population cells from hepatocellular carcinoma cells
Jing Fan, Ren Li, Rui Zhang, Hai-Liang Liu, Ning Zhang, Fu-Qing Zhang, Ke-Feng Dou
Jing Fan, Ren Li, Hai-Liang Liu, Ning Zhang, Fu-Qing Zhang, Ke-Feng Dou, Department of Hepatobiliary Surgery, Xijing Hospital, the Fourth Military Medical University, Xi’an 710032, Shaanxi Province, China
Rui Zhang, Department of Biochemistry and Molecular Biology, the Fourth Military Medical University, Xi’an 710032, Shaanxi Province, China
Author contributions: All authors contributed equally to the work.
Correspondence to: Ke-Feng Dou, Department of Hepatobiliary Surgery, Xijing Hospital, the Fourth Military Medical University, 17 Changle Western Road, Xi'an 710032, Shaanxi Province, China. doukef@fmmu.edu.cn
Telephone: +86-29-84775255 Fax: +86-29-84775255
Received: February 25, 2007
Revised: August 6, 2007
Accepted: October 26, 2007
Published online: December 7, 2007
Abstract

AIM: To understand the role and significance of side population (SP) cells from hepatocellular carcinoma (HCC) in hepatocarcinogenesis, development, relapse and metastasis, we simulated the denutrition conditions that cancer cells experience in clinical therapy, observed the different anti-apoptosis ability of SP cells and non-SP cells under such conditions, and established the possible effects of P53, Bcl-2 and Bax on survival of SP cells.

METHODS: We used flow cytometry to analyze and sort the SP and non-SP cells in established HCC lines MHCC97 and hHCC. We evaluated cell proliferation by methyl thiazolyl tetrazolium (MTT) assay and investigated the expression of p53, bcl-2 and bax genes during denutrition, by RT-PCR and immunofluorescence staining.

RESULTS: The percentage of SP cells in the two established HCC lines was 0.25% and 0.5%, respectively. SP cells had greater anti-apoptosis and proliferation ability than non-SP cells. Expression of Bcl-2 and Bax in SP and non-SP cells differed during denutrition. The former was up-regulated in SP cells, and the latter was up-regulated in non-SP cells.

CONCLUSION: It may be that different upstream molecules acted and led to different expression levels of Bcl-2 and Bax in these two cell lines. There was a direct relationship between up-regulation of Bcl-2 and down-regulation of Bax and higher anti-apoptosis ability in SP cells. It may be that the existence and activity of SP cells are partly responsible for some of the clinical phenomena which are seen in HCC, such as relapse or metastasis. Further research on SP cells may have potential applications in the field of anticancer therapy.

Keywords: Side population; Hepatocellular carcinoma; Bax; Bcl-2; Apoptosis