Effects And Mechanism Of Schisandrin B On Proliferation And Migration Of Human Cervical Cancer Siha Cells

Objective: Cervical cancer is a kind of multiple and common malignant tumor of gynecological reproductive system,which seriously damages and endangers women’s health.Cervical cancer is a frequent and common gynecological malignant tumor of the reproductive system,which seriously damages the health of women.The occurrence of cervical cancer is the result of the comprehensive factors.To date,no specific drugs have been developed to effectively combat cervical cancer.Schisandrin B(Sch B)is a kind of traditional Chinese medicine,which also plays a role in anti-inflammatory,anti-oxidation,inhibition of multidrug resistance and anti-tumor activity.Schisandrin B is one of the active components of Schisandrin B with many effects.Currently,there are few studies on the inhibitory effect of Schisandrin B on Siha cells of cervical squamous cell carcinoma and its mechanism.The purpose of this study was to explore the effect of Schisandrin B on the proliferation and migration of human cervical cancer Siha cells and its mechanism,so as to provide a basis for the study on the anti-tumor effect of traditional Chinese medicine.Methods: Gene Expression Omnibus(GEO)database was used to summarize and analyze the differential genes between squamous cervical cancer and normal cervical tissue.KEGG(Kyoto Encyclopedia of Genes and Genomes)and GO(Gene Ontology)enrichment pathways were analyzed according to the screening results.The gene expression profile of PKM2 was analyzed by The Human Protein Atlas database.Siha cells in logarithmic growth phase were divided into blank control group and Schisandrin B treatment group with different concentrations.The blank control group was the group without any treatment.MTT colorimetry was used to determine the inhibitory effect of Schisandrin B on the growth and proliferation of Siha cells treated with different concentrations;The effect of Schisandrin B on Siha cell migration was detected by scratch test;Real-time fluorescent quantitative PCR and Western blot method to detect Schisandrin B element applies to Siha cells after pyruvate kinase 2(PKM2),signal transduction and transcriptional activation factor 3(signal transducers and activators of transcription,STAT3),cell cycle protein D1(Cyclin D1),the BCL-xl apoptosis protein,MMP-2(matrix metalloproteinases 2)and MMP-9(matrix metalloproteinases 9)m RNA and protein expression;The glucose intake and lactic acid production of Siha cells treated with Schisandrin B chinensis ethylin for 48 h were determined by the method of glucose and lactic acid content determination.Results: The results showed that 2382 differentially expressed genes were obtained by screening,and PKM gene was included in the up-regulated genes.In the KEGG enrichment pathway,21 up-regulated genes were enriched and 11down-regulated genes were enriched.The results of gene expression profilation showed that PKM2 gene was highly expressed in Siha cells compared with that in normal cervix.MTT results showed that the malignant proliferation of Siha cells was inhibited after 48 h treatment with Schisandrin B.The inhibitory effect was gradually enhanced with the increase of drug concentration(25 μmol/L,50 μmol/L,100 μmol/L,all P<0.05);The results of scratch experiment showed that Schisandrin B treated Siha cells for 48 h reduced the migration ability of Siha cells.The higher the drug concentration,the lower the mobility.The migration difference of high concentration group was significant.(P<0.01);Western blot results showed that STAT3,PKM2,Cyclin D1,and BCL-xl were inhibited by the drug compared with the untreated group(low,medium and high concentration groups p<0.05),MMP-2(medium and high concentration group P<0.05),MMP-9(medium and high concentration group P<0.05)the relative expression level of protein was down-regulated;Real-time PCR results showed that the m RNA levels of STAT3,PKM2,Cyclin D1,BCL-xl,MMP-2,and MMP-9 in different concentrations of treated groups were decreased compared with those in non-treated groups.Conclusion: Bioinformatics analysis showed that the occurrence and development of cervical cancer may be closely related to PKM gene.Schisandrin B inhibited the proliferation and migration of Siha cells in HPV16 positive cervical squamous cell carcinoma.The mechanism of Schisandrin B action may inhibit cell growth and reproduction by down-regulating the level of PKM2 in the glycolytic pathway,thereby affecting the phosphorylation of STAT3,and reducing the expression of Cyclin D1 and anti-apoptotic protein BCL-xl,as its downstream targets.Schisandrin B inhibit cell growth and reproduction by down-regulating the level of PKM2 in the glycolytic pathway,thereby affecting the phosphorylation of STAT3,and reducing the expression of Cyclin D1 and anti-apoptotic protein BCL-xl,which are its downstream cycle affecting targets.The mechanism of the action of Schisandrin B reduce the cell migration ability by down-regulating the expression of MMP-2 and MMP-9 and thus reducing the degradation of extracellular matrix,thus exerting the anticancer effect.