Mechanism Of Glaucocalyxin B Induced Proliferation Inhabitation To HeLa And SiHa Of Cervical Cancer

Background Cervical cancer is the third most common cancer among women worldwide,with the fourth highest fatality rates among females.In particular,it is still a severe health issue in developing countries,although the incidence of this disease is strongly decreasing in developed countries due to early diagnosis.Currently,consist primarily of radiotherapy combined with platinum-based chemotherapy is standard of care for advanced cervical cancer.Although the addition of platin-based chemotherapy to primary radiotherapy has increased 5-year survival of advanced-stage cervical cancer patients,the high-dose use of chemotherapeutic drugs is limited due to their systemic toxicity.Therefore,additional therapies for this deadly disease are critical.Complementary and alternative medicines can perhaps benefit cervical cancer patients as an adjunctive therapy.Of all medicines,Chinese traditional medicine has become increasing prominent and popular in cancer patients due to its efficacy and low toxicity?Glaucocalyxin,a ent-kauranoid diterpenoid isolated from the Chinese traditional medicine ‘Rabdosia japonica',has been used in traditional medicine as an antibacterial,anti-inflammatory,and anti-cancer agent.Glaucocalyxin has three major forms,Glaucocalyxin A,B and C.Recently Glaucocalyxin A and B,has attracted great attention for its potent anticancer effects,but its potential molecular targets involved in tumorigenesis have not been reported.Objective The aim of this study is to investigate the effects and potential molecular mechanism of Glaucocalyxin B to cervical carcinoma Hela?Siha cells.Method1)Culture cervical carcinoma cell(Hela?Siha)in virto,then treated with different concentrations of GLB for different times,using MTT method to dectet the cell viability.2)Apoptosis rate Hela?Siha cell were treated with different concentrations of GLB for 24 h,then detect the apoptosis rate by flow cytometry(PI and Annexin V-FITC staining);3)The relative protein expressions of PTEN ? P-Akt ? PARP ? Beclin1 ?LC3?/?were assayed by western blot.Result1)Test result of cells proliferation inhabitation ratio by MTT colorimetry The MTT colorimetry showed that the proliferation ability of Cervical Cancer cells declined after treated with Glaucocalyxin B than untreated group(P<0.05);The declined rate of proliferation rates of He La and Si Ha cells treated with different concentrations of Glaucocalyxin B were significant different(P<0.05),which presented a dose and time dependent.2)Cell apoptosis rate detected by Annexin V-FITC/ PI flow cytometry After dealt with different concentrations of Glaucocalyxin B for 24 h,the apoptosis rates of He La and Si Ha cells were increased significantly as the concentrations increased compared with control group respectively.3)The relatively amount of expression of PTEN?P-AKT?PARP from PI3K/Akt signal pathway and Beclin1?LC3?/ LC3?detected by western-bloting.The relative expression of P-AKT and Beclin1 in Hela and Si Ha cells after treated with Glaucocalyxin B were declined greatly compared to the control group(P<0.05).The expression of PTEN ?C-PARP and LC3?/ LC3?increased compared to the control group(P<0.05).Conclusion Glaucocalyxin B can inhabit the proliferation of Hela?Siha cell,which is relative with concentration and time,inducing Hela?Siha cell happen apoptosis and autophagy.The mechanism is related with negative-regulation of PI3K/AKT signal pathway,and Beclin1 may regulate apoptosis and autophagy.