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The Effect of Notch1 Gene Transfection on the Growth of Hep3B Hepatocarcinoma Cells and the Related Mechanisms
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Abstract:
Objective: To investigate the effects of Notch1 gene transfection on the growth of human hepatocarcinoma cells and to explore the mechanisms of Notch signaling. Methods: Recombinant retrovirus MSCV-ICN/GFP and MSCV-GFP were prepared by calcium phosphate precipitation mediated gene transduction of 293T package cells. The recombinant retroviruses were used to infect human hepatocarcinoma cells Hep3B. After infected, proliferation of Hep3B cells was observed by MTT assay. Cell cycle distribution was analyzed by FACS and p53 expression was detected by Western blot. Results: After infection of MSCV-ICN/GFP recombinant retrovirus, the growth of Hep3B cells was inhibited significantly and cell cycle distribution analysis showed that Hep3B cells in G0/G1 phase were increased. Western immunoblotting showed that Notch1 signaling up regulated p53 expression. Conclusion: Notch1 activation could inhibit the growth of hepatocarcinoma cells through G0/G1 cell cycle arrest and p53 induction.
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