Effects Of Glaucocalyxin A On Cervical Carcinoma Hela、Siha Cell And Its Mechanism

Objective: To investigate the effects of Glaucocalyxin A on cervical carcinoma Hela、Siha cell,and the molecular mechanism.Methods: Cervical carcinoma cell (Hela、 Siha) were treated with differentconcentrations of GLA, after different times of GLA treatment, the cell viability wasmeasured by MTT method.Apoptosis and cell cycle of Hela、Siha cell after48h ofdifferent concentrations of GLA exposure were detected by flow cytometry （PI andAnnexinV-FITC staining）; The morphological change of Hela、Siha cell was observed byWright＇ s-Giemsa stain; The ultrastructure of Hela、 Siha cell was examined bytransmission electron microscopy. The protein expressions of Bax、Bcl-2、Caspase3、Caspase9、LC3、Beclin1were assayed by western blot.Results:(1)MTT assay showed that after treated with different concentrations ofGLA and different time,GLA had a significant anti-proliferation effect on cervicalcarcinoma cell in a dose-dependant and time-dependant manner.(2) The Morphologicalchanges of Hela、Siha cell were detected by Wright＇s-Giemsa stain.The results showedthat after treated with different concentrations of GLA,the numbers of cellreduced,turned into the round shape and began to shed off.(3) Cell cycle arrest wasexamined by Flow Cytometry (FCM). After treated with different concentrations of GLAfor48hours, the S phase rates of cervical carcinoma Hela、Siha cell were increased.(4)AnnexinV-FITC staining detects apoptosis rates of cervical carcinoma Hela、Siha celltreated with different concentrations of GLA for48hours were increased（.5）After treatedwith3+MA and GLA,the apoptasis rate of cervical carcinoma Hela、Siha cell wasincreased according to treated with GLA for24h.(6) After treated with differentconcentrations of GLA for24h, the protein of bax was upregulated, bcl-2was downregulated,Caspase3was upregulated, Caspase9was upregulated by western blot.They were also in a dose-dependant manner（P<0.05）.Conclusion GLA can inhibit the proliferation of Hela、Siha cell in a concentration-dependent and time-dependent manner, induce Hela、Siha cell to apoptosis remarkably. Themitochondrial signal transduction pathway was involved in its mechanism.