Mechanism Research Of Glaucocalyxin B-induced Growth Inhibition,apoptosis And Autophagy Of Cervical Cancer Cells By Increasing ROS

BackgroundCervical cancer incidence and mortality are increasing year by year and tend to be younger,and it is an important disease that endangers women’s health and life.Surgical treatment combined with radiotherapy and chemotherapy is an important treatment for cervical cancer patients.During chemotherapy,there will be serious side effects and drug resistance caused by long-term use.This makes the treatment of cervical cancer less than expected.The low-toxicity and high-efficiency antitumor properties of traditional Chinese medicine can avoid most side effects during cancer treatment.Glaucocalyxin B is an anti-tumor and other biological activity,which has been confirmed to inhibit the proliferation of cervical cancer Hela and Si Ha cells,and induce their apoptosis and autophagy.ROS（reactive oxygen species）are highly reactive oxygen species,and excessive accumulation can lead to cell death.Therefore,increasing the level of ROS in tumor cells has become a promising anti-tumor strategy.It is still unclear whether the inhibitory effect of Glaucocalyxin B on cervical cancer is related to ROS.ObjectiveIn this thesis,we investigated the effects of Glaucocalyxin B on the proliferation,apoptosis and autophagy of cervical cancer C33 A cells,explored whether Glaucocalyxin B played a regulatory role in intracellular ROS in cervical cancer cells,and revealed that ROS played a role in the regulation of cervical cancer cells by Glaucocalyxin B.Roles in cell proliferation,apoptosis and autophagy.Methods1.CCK-8,plate clone formation and Western blot experiments were used to analyze the survival rate,clone formation ability,apoptosis and autophagy-related proteins of cervical cancer C33 A cells after adding Glaucocalyxin B respectively;2.Determination of ROS levels in cervical cancer C33 A,Hela and Si Ha cells after treatment with Glaucocalyxin B by ROS detection kit;3.The survival rate and clone formation of cervical cancer C33 A,Hela and Si Ha cells were analyzed by CCK-8,plate colony formation and Western blot experiments respectively in the presence of ROS inhibitor N-acetylcysteine after adding Glaucocalyxin B capacity and changes in apoptosis-and autophagy-related proteins.Results1.The survival rate and clonogenesis ability of cervical cancer C33 A cells were decreased,and the contents of apoptosis-related protein CL-PARP and autophagy related protein LC3 were increased after the addition of Glaucocalyxin B.2.The intracellular ROS levels of cervical cancer C33 A,Hela and Si Ha cells increased gradually after Glaucocalyxin B treatment.3.In the presence of ROS inhibitor N-acetylcysteine,the survival rate and clonogenesis ability of cervical cancer C33 A,Hela and Si Ha cells were increased,and the contents of apoptosis-related protein CL-PARP and autophagy related protein LC3 were decreased by Glaucocalyxin B.ConclusionGlaucocalyxin B inhibits cervical cancer cell proliferation and induces apoptosis and autophagy by increasing the level of intracellular ROS.