Proliferation Inhibition And Apoptosis In Human Cervical Cancer Hela Cells Induced By JH42-1in Vitro

Objective To investigate the effect of proliferation inhibition and apoptotic induction in HeLa cells induced by JH42-1(Isopropenyl butene dioic acid mono-methyl ester), a novel chemical, for the purpose of developing JH42-1into a new drug for treating human cervical cancer. Methods Human cervical cancer HeLa cells were treated by JH42-1at various concentrations. The morphological changes of the HeLa cells after treatments were observed by microscopy. The effects of proliferation inhibition induced by the chemical were measured with MTT (Methyl thiazolyl tetrazolium) assay. Apoptotic cells induced by JH42-1and stained by AnnexinV-EGFP/PI were quantified by flow cytometry, and apoptotic ratios were calculated. Furthermore, apoptotic cells stained by AnnexinV-EGFP/PI and Hoechst-33258respectively were investigated under fluorescence microscopy. Results1.Morphological changes, including cell shrinkage and nuclear condensation, occurred in the apoptotic cells treated by the chemical, which did not appear in the negative control group. These changes were obvious in the cells treated at the concentration of30μmol/1for24hours. With the increase of concentration and the hours, the morphological changes were much more obvious. Meanwhile, a large number of cells were floating in the media.2. Calculated by MTT assay, ratios of proliferation inhibition in the cells treated for24hours increased. The proliferation inhibition rates in the groups treated by30μmol/1JH42-1and greater concentrations were higher than that in negative control groups significantly (p<0.05), in a concentration dependent manner.3. Treated by JH42-1for24,48,72hours, apoptosis was induced. The rates of apoptotic cells in30μmol/1and above groups for24hours and in20μmol/1and above groups for48,72hours increased significantly (P<0.05), in a concentration dependent manner, compared with that in negative control group.4. By fluorescence microscopy, both the early and the late apoptotic cells stained by AnnexinV-EGFP/PI could be seen in groups at various concentrations for72hours. Stained by Hoechst-33258, the morphology changes of apoptosis were observed, and the percentage of apoptotic cells increased with increase of concentration. Conclusion Based on the evidences above, we can come to a conclusion that JH42-1inhibits proliferation and induces apoptosis in HeLa cells. JH42-1is a promising novel anticancer drug for treating human cervical cancer.