Objective        To determine the inhibitory effect of a recombinant adenovirus of human p66Shc (AdenoⅩ-p66Shc) on the proliferation of human cervical cancer HeLa cells and investigate the possible underlying mechanism.       Methods        The Hela cells were infected with AdenoⅩ-LacZ and AdenoⅩ-p66Shc respectively. The cells without infection served as blank control. Cell viability was analyzed by MTT assay. Reactive oxygen species (ROS) was measured by DCFH-DA fluorescent probes and flow cytometry. The levels of 8-OHdG were detected by ELISA assay. Cell apoptosis was detected by Annexin Ⅴ-FITC/PI staining. Western blotting was used to observe the expression of associated proteins. Co-immunoprecipitation (Co-IP) was used to investigate the possible interaction between p66Shc and p53.       Results        MTT assay showed that the proliferation of HeLa cells was significantly suppressed (P0.05) by AdenoⅩ-p66Shc in a dose-dependent manner

and its inhibition could be blocked by N-acetyl-L-cysteine. Meanwhile

the levels of ROS and 8-OHdG were significantly increased after AdenoⅩ-p66Shc infection (P0.05). The expression of p53

Cyclin B1 and p-p53 was up-regulated with the over-expression of p66Shc

but the result of Co-IP demonstrated that p66Shc and p53 were not directly combined

suggesting the expression and activity of p53 was regulated by other molecular activated by p66Shc.       Conclusion        AdenoⅩ-p66Shc induces ROS generation and DNA oxidative damage

and up-regulates the expression of p53 and p-p53

and thereby inhibits the proliferation of HeLa cells.