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World J Gastroenterol. Jun 21, 2008; 14(23): 3754-3758
Published online Jun 21, 2008. doi: 10.3748/wjg.14.3754
Effect of fragile histidine triad gene transduction on proliferation and apoptosis of human hepatocellular carcinoma cells
Rong-Hua Xu, Liang-Yan Zheng, Dong-Lei He, Jian Tong, Li-Ping Zheng, Wu-Ping Zheng, Jin Meng, Li-Ping Xia, Cong-Jun Wang, Ji-Lin Yi
Rong-Hua Xu, Liang-Yan Zheng, Dong-Lei He, Jian Tong, Li-Ping Zheng, Wu-Ping Zheng, Jin Meng, Li-Ping Xia, The Affiliated Hospital of Hainan Medical College, Haikou 570102, Hainan Province, China
Cong-Jun Wang, Department of General Surgery, First People’s Hospital Affiliated to Shanghai Jiaotong University, Shanghai 200080, China
Ji-Lin Yi, Department of General surgery, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan 430030, Hubei Province, China
Author contributions: Xu RH and Zheng LY wrote the paper and organized the figures; Xu RH, He DL, Tong J, Zheng LP, Zheng WP, Meng J, Xia LP did cell culture, transduction, RT-PCR, Western blot and animal experiments; Wang CJ carried out the statistical analysis; Yi JL supervised the writing and organization process.
Correspondence to: Dr. Rong-Hua Xu, Department of Oncology Surgery, The Affiliated Hospital of Hainan Medical College, Haikou 570102, Hainan Province, China. xu_ronghua2231@hotmail.com
Telephone: +86-898-66528115
Received: October 29, 2007
Revised: April 19, 2008
Accepted: April 26, 2008
Published online: June 21, 2008
Abstract

AIM: To evaluate the inhibitory effects of human fragile histidine triad (FHIT) gene on cell proliferation and apoptosis in human hepatocellular carcinoma line Hep3B in vitro.

METHODS: A recombinant pcDNA3.1 (+)/FHIT including the functional region of FHIT gene was constructed and transferred into human hepatocellular carcinoma cells in vitro. mRNA and protein expression of the FHIT gene in the transfected cells was detected by RT-PCR and Western blot, respectively. The effect of FHIT on proliferation was detected by MTT assay. Changes in cell cycle and apoptosis were assayed by flow cytometry. Five mice received subcutaneous transplantation of Hep3B-FHIT; 5 mice received subcutaneous transplantation of normal Hep3B and Hep3B-C as controls. The body weight of nude mice and tumor growth were measured.

RESULTS: RT-PCR and Western blot analysis showed that the expression level of FHIT-mRNA and FHIT protein was higher in Hep3B cells after infection with pcDNA3.1 (+)/FHIT. The growth of Hep3B cells treated with pcDNA3.1 (+)/FHIT was significantly inhibited. The pcDNA3.1 (+)/FHIT-transfected Hep3B cells showed a significantly higher cell rate at G0-G1 phase and increased apoptosis in comparison with controls (P < 0.05). The growth of transplanted tumor was inhibited markedly by FHIT. Tumors arising from the Hep3B-FHIT cells occurred much later than those arising from the Hep3B and Hep3B-C cells. The growth of Hep3B-FHIT cells was slow and the tumor volume was low.

CONCLUSION: Transduction of FHIT gene inhibits the growth of human hepatocellular carcinoma cells and induces cell apoptosis in vivo and in vitro.

Keywords: Hepatocellular carcinoma, Gene therapy, Fragile histidine triad gene