Trichosanthin Induced Apoptosis In Hela Cells And Its Mechanism Study

Objective : study Trichosanthin induced apoptosis in Hela cells in vitro so as to find a new and effective drug which can inhibit the cervical cancer cells's growth by apotosis; explain the mechanism in the molecular lever to lay scientific foundations for Trichosanthin's clinical use in the therapy of cervical cancer.Methods: Hela cells had been used to testify the apoptosis induced by Trichosanthin with the methods of morphological observation, MTT assay, DNA agarose gel electrophoresis and FCM techniques. Western blot and DNA microarray had been used to obtain the gene expression panel of Hela cells before and after treated with Trichosanthin .Results: after treated with Trichosanthin , typical apoptotic changes, such as cell volume minishing , cytoplasm condensing , refraction rate increasing and some adherent cells transitting to suspension cells, have take place by the morphological observation; MTT assay showed that Hela cell's growth were inhibited after treated with 20ug/ml's Trichosanthin for 48 hours and Hela cells's energy decreased with the concentration increase of Trichosanthin (within the concentration of 20ug/ml to 120ug/ml); Hela cells'DNA were broken up into 180-200bp and DNA ladder were observed by Agarose electrophoresis ; sub-G1 peak in the DNA contents frequency histograms had also been observed by FCM technique; bcl-2's expression descended but bax's expression enhanced with Trichosanthin's concentration increase according to the Western blot ; in the DNA microarray test , there were 78 pieces of genes changed distinctly, 16 genes associated with the adhesion and interaction between cells were down-regulated and 62 genes (including the genes nearly related with apoptosis such as BAX,CASP9,DFFB,P53 ect )were up-regulated.Conclusion: Trichosanthin can inhibit the growth of hela cells in vitro and this effect results from Trichosanthin induced apoptosis. the possible molecular mechanism of Trichosanthin induced apoptosis are as follows: (1) stimulation→bcl-2/bax→cytochrome c→caspase-3→DFF: bcl-2's expression decreased but bax's expression...