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| Apoptosis-inducing Effect of Anti-p73 Gene on Human Colon Cancer Cell HepG2 |
| 302 Military Hospital of China,Beijing 100039,China |
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Abstract Objective:To discuss the mechanism of p73 inducing tumor cells into apoptosis by constructing recombinant expression plasmid pBP73 and observing the expression and apoptosis-inducing effect of anti-p73 gene on colon cancer cell HepG2 Methods: Anti-p73 gene was cloned into the plasmid pBabe to form the recombinant plasmid pBP73, and then pBP73 was transformed into HepG2 cells and then agarose electrophoresis and flow cytometry were used to verify apoptosis of tumor cells. At last caspase-3 was detected by Western blotting. Results: Sequence analysis justified the recombination of plasmid pBP73. Virus could be collected from cultured supernatant of infected 293A cells with the titer of 5×107pfu. And 48h after infection, agarose electrophoresis of genomic DNA showed typical ladder-like pattern and flow cytometry analysis showed obvious apoptosis peaks with the highest percentage rate of apoptotic cells present and cellular caspase-3 expression could be seen in pBP73 transformed group. Conclusion: Anti-p73 gene could induce apoptosis in human colon cancer cells HepG2 in vivo by activating caspase-3 expression.
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Received: 09 November 2009
Published: 26 February 2010
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