Study On Apoptosis Mechanism Of Cervical Cancer He La Cells Induced By PI3K/AKT Inhibitor LY294002 Combined With Shikonin

ObjectiveCervical cancer,one of the most common malignant tumors in women,is seriously threatens the life and health of patients.Now,surgical resection,chemotherapy,radiotherapy and others methods are mainly used for the clinical treatment of cervical cancer.However,patients often experience postoperative recurrence,poor efficacy of chemotherapy and radiotherapy,or severe side effects of drugs.Therefore,it is urgent to find some effective drugs with low side effects in clinical treatment.Chinese herbal medicines have been studied and applied in China since ancient times.Numerous studies have shown that many traditional Chinese medicines have good inhibitory effects on tumors,and shikonin is one of them.Shikonin plays an important role in inducing apoptosis of various tumor cells,including colorectal cancer,ovarian cancer,gastric cancer,skin cancer and cervical cancer,which have obvious proliferation effect and low toxic and side effects.In this study,PI3K/AKT inhibitor LY294002 combined with shikonin was used to treat cervical cancer He La cells cultured in vitro,to investigate its mechanism of action on apoptosis of cervical cancer cells.MethodsHe La cells were treated with different concentrations of shikonin to observe the morphological changes.The activity of He La cells was determined by cck-8 assay.The effects of shikonin on He La cell cycle,apoptosis and ROS were determined by cell flow assay.Western blot was used to detect the effect of shikonin on He La cell related protein expression.Finally,the inhibitor LY294002 was combined with shikonin to detect the effect of the combination on the level of He La cell apoptosis.Results 1.shikonin significantly inhibit the proliferation of He La cells in a time-concentration dependent manner.He La stimulated by shikonin will underwent morphological changes and reduce cell viability.2.shikonin reduce the expression of cyclin-d1 protein in He La cells and block the cells from remaining in G0/G1 phase.shikonin significantly increase the apoptosis level of He La cells.3.Stimulation with shikonin increase the intracellular ROS level of He La cells.4.When shikonin stimulates He La cells,it activates the AKT signaling pathway,triggering the stress response and protective autophagy of the cells to resist the drug effect.5.The combination of PI3K/AKT inhibitor LY294002 with shikonin significantly increase the sensitivity of He La cells to shikonin and induce its apoptosis.ConclusionShikonin has a significant inhibitory effect on He La cells of cervical cancer.shikonin can inhibit the proliferation and growth of He La cells and induce their apoptosis by reducing the activity of He La cells,stasis their cell cycle at G0/G1 phase,and increase the intracellular ROS level.When stimulated by shikonin,He La cells can activate the AKT signaling pathway,and generate stress response and protective autophagy.When the PI3K/AKT inhibitor LY294002 was used to inhibit the activity of AKT,the sensitivity of He La to shikonin could be enhanced.The combination of the two can significantly induce the apoptosis of He La cells.