[Recombinant adenovirus vectors carrying antisense MMP2 inhibit invasion of HCC cells in vitro]

Ming-Man Zhang, Xing-Hua Gou, De-Hua Li, Lü-Nan Yan, Dong Xia, Lei Han, Zhi Su, Lai-Ying Zhao, Hai-Yang Hu
Zhonghua Gan Zang Bing za Zhi, Zhonghua Ganzangbing Zazhi, Chinese Journal of Hepatology 2005, 13 (4): 267-70

OBJECTIVES: To construct a recombinant adenoviral vector carrying antisense matrix metalloproteinase-2 (MMP2) and to study its inhibitory effects on the invasiveness and migratory capacity of hepatocellular carcinoma (HCC) cell line HepG2 in vitro.

METHODS: Total RNA was extracted from HCC. Then a 500 bp fragment at the 5' end of the human MMP2 cDNA sequence was synthesized by polymerase chain reaction (PCR) and was reversely inserted into the multiclone site (MCS) of the shuttle plasmid pAdTrack-CMV. With the resultant plasmid and the backbone plasmid pAdEasy-1, the homologous recombination took place in the E.coli BJ5183 and the recombinant adenoviral plasmid carrying the antisense MMP2 gene was constructed. The adenovirus (Ad-MMP2AS) was packaged and amplified in the HEK 293 cells and the viral titer was checked by GFP. Using the Boyden chamber model, the influence of Ad-MMP2AS on the invasion ability of HepG2 cells was determined in vitro.

RESULTS: The recombinant adenovirus vector carrying antisense MMP2 was constructed successfully and a strong green fluorescence was observed in HepG2 cells under a fluorescence microscope. The viral titer was 1 x 10(8); Ad-MMP2AS can effectively inhibit the penetrating capacity of HepG2 cells through Matrigel in vitro.

CONCLUSION: The recombinant adenovirus with antisense MMP2 can effectively inhibit the invasiveness and migratory capacity of HepG2 in vitro and may have potential in treating HCC.

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