Experimental Study On Inhibition Of Growth And Induction Of Apoptosis Of Cervical Cancer Hela Cells By 5, 7-Dimethorychrysin In Vitro

Objective: The aim of this study was to investigate the effect of 5,7-dimethorychysin (dMChR)on cell proliferation and cell apoptosis of cervical cancer Hela cells in vitro and its mechanism.Methods: Cervical cancer Hela cells were cultured in vitro.The antiproliferation effect of dMChR on Hela Cells was detected by MTT assay, and take Human umbilical vein endothelial ECV-304 cell line as nomal control group; The ability of clone formation of Hela cells was evaluated by clone formation test; Cell apoptosis morphologic changes of Hela cells were investigated by fluorescence using AO/EB double staining. The apoptosis rate of Hela cells was determined by flow cytometry analysis after PI staining; The expression of Bcl-2,Bax and Caspase-3 protein of Hela cells were investigated by indirect immunifaction technique using FITC fluorescence tagging.Results:1.MTT assay indicated dMChR(0.3μM, 3μM, 30μM, 100μM) inhibited the proliferation of Hela cells for 48h in a dose-dependent manner,and its IC50 was 30.09μM; while the proliferation of ECV-304 cells was a little affected at the same concentration, IC₅₀ of dMChR to ECV-304 cells was 15415.8μM. SI of dMChR to Hela cells was 512.32. These results of colony formation test that dMChR(0.3μM, 3μM, 30μM, 100μM)treated Hela cells for 7 days were consistent with that of MTT assay test. 2. Fluorescence using AO/EB double staining and flow cytometry analysis after PI staining showed the apoptosis of Hela cells were induced by dMChR in a dose and time dependent manner. 3.The result of indirect immunifaction technique using FITC fluorescence tagging indicated that not only the expression of Bcl-2 protein decreased and the expression of Bax,Caspase-3 protein increased in Hela cells treated with dMChR(3μM, 30μM, 100μM)for 24 hours, but also the expression of Bcl-2 protein decreased and the expression of Bax,Caspase-3 protein increased in Hela cells treated with dMChR(30μM)for 12 hours, 24 hours. dMChR (100μM) and ChR(100μM) could decrease the expression of Bcl-2 protein and up-regulate of Bax, Caspase-3 protein in Hela cells, but the effect of dMChR was stronger that of ChR. 4.Those tests all indicated that the antiproliferation and apoptosis induction effect of dMChR (100μM) on Hela cells were stronger than ChR(100μM).Conclusions:1. The present study results suggest that 5, 7-dimethorychysin can significantly inhibit the proliferation of Hela cells , but its antiproliferation effect on ECV-304 cells is weaker.2. 5, 7-dimethorychysin can inhibit the proliferation of cervical cancer Hela cells and induce the apoptosis of cervical cancer Hela cells in a dose-dependent and time-dependent manner.3. The apoptosis induction effect of dMChR on Hela cells are possibly associated with upregulating ration of Bax / Bcl-2 and activation expression of Caspase-3 protein.4. The antiproliferation and apoptosis induction effect of dMChR on Hela cells is possibly stronger than ChR, and their effect mechanism on Hela cells are possibly resemble.