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Overexpression of TIMP-1 mediated by recombinant adenovirus in hepatocellular carcinoma cells inhibits proliferation and invasion in vitro |
Dong Xia, Lu-Nan Yan, Jian-Guo Xie, Yu Tong, Mao-Lin Yan, Xin-Ping Wang, Ming-Man Zhang and Lan-Ying Zhao |
Chengdu, China
Author Affiliations: Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041, China (Xia D, Yan LN, Xie JG, Yan ML, Wang XP and Zhang MM); Department of General Surgery, Luzhou Medical College, Luzhou 646000, China (Xia D); and Genetic Engineering Laboratory, Chengdu Di'ao Group Co., Ltd. (Tong Y and Zhao LY), Chengdu 610041, China
Corresponding Author: Lu-Nan Yan, MD, Department of General Surgery, West China Hospital, Sichuan University, Chengdu 610041, China (Tel: 86-28-85422477; Fax: 86-28-85423724; Email: YanLu-Nan@hotmail.com) |
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Abstract BACKGROUND: Matrix metalloproteinases (MMPs) and its natural tissue inhibitors of metalloproteinases (TIMPs) are involved in cancer progression. This study was undertaken to determine the effects of overexpression of TIMP-1 on human hepatocellular carcinoma (HCC) cell growth, proliferation, and invasion.
METHODS: Employing the efficient AdEasyTM system, recombinant adenovirus AdTIMP-1 containing full-length cDNA of TIMP-1 was generated by homologous recombination and amplified in 293 cells. Then, human HCC cell line (HepG2) underwent gene transfection to overexpress TIMP-1 (so-called HepG-T cells). The mRNA and protein expressions of TIMP-1 were detected with RT-PCR and Western blotting, respectively. The ultrastructure was observed with a transmission electron microscope and the proliferation of HepG-T cells was determined by MTT assay and growth curve. The potential of in vitro invasion was measured with Millicell Chamber.
RESULTS: The resulting AdTIMP-1 and HepG-T cells were generated and the expression of TIMP-1 was detected in vitro. The cell proliferation curves and MTT assay showed HepG-T cells' growth, and proliferation were obviously inhibited. The invasion across Matrigel-coated filters was significantly decreased compared with controls. The suppression rate of HepG-2 cells with AdhTIMP-1 transfection was 50%, and AdhTIMP-1 transfection inhibited by more than 91.6% of the invasion into the Matrigel-coated filter (P<0.01).
CONCLUSIONS: TIMP-1 overexpression results in the suppression of proliferative and invasive potential of HepG2 cells in vitro. This study demonstrates the potential role of TIMP-1 as a target for liver cancer gene therapy and has laid a foundation for further study on its anticancer function.
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Cite this article: |
Xia D,
Yan LN,
Xie JG,
et al.
Overexpression of TIMP-1 mediated by recombinant adenovirus in hepatocellular carcinoma cells inhibits proliferation and invasion in vitro.
Hepatobiliary Pancreat Dis Int
2006;
5(3):
409-415. DOI:
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URL: |
http://dx.doi.org/ OR http://www.hbpdint.com/EN/Y2006/V5/I3/409 |
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