The Inhibitory Effect And Molecular Mechanism Of Polyphyllin Ⅰ In Cervical Cancer

Cervical cancer(Cervical Cancer,CC)is the fourth most common cancer in women in the world.In 2020,GLOBOCAN estimated that 604,000 women were diagnosed with cervical cancer,and 342,000 women died due to the disease.In the clinical subsequent treatment of cervical cancer,radiotherapy and chemotherapy is currently considered as the standard treatment for patients with advanced/recurrence.However,the drug resistance of cancer cells and the characteristics of easy metastasis are one of the important reasons for the deterioration and even death of cancer patients.Therefore,the molecular mechanism of chemoresistance in cervical cancer is urgent.Previous studies show that the effective component of traditional Chinese medicine chonglou(Polyphyllin)has a certain inhibitory effect on tumor growth in lung cancer,gastric cancer,breast cancer,rectal cancer,breast cancer and other solid tumors.However,there are few studies in cervical cancer.This study explores the inhibitory effect and related action mechanism of Polyphyllin Ⅰ on cervical cancer resistant cells,and aims to provide a new therapeutic means for the treatment of cisplatin-resistant cervical cancer patients.Purpose:1.Cell experiments were performed to verify the effect of Polyphyllin Ⅰ on cervical cancer resistant cells.2.Relevant experiments were conducted to verify the mechanism of Polyphyllin Ⅰ enhancing cisplatin chemotherapy sensitivity.Method:1.HeLa and SiHa primary cells and cisplatin-resistant cervical cancer cell lines(HeLa/DDP and SiHa/DDP)were cultured to study the effect of chonglou Polyphyllin Ⅰ on cervical cancer resistant cells through cell proliferation,cell cycle,cell apoptosis,cell migration,cell cloning and other experiments.The expression of P53,P21,Bax and Bcl-2 proteins was measured by Western-blot to verify whether Polyphyllin Ⅰ acts against the P53 and P21 pathway by affecting Bax and Bcl-2 and inducing apoptosis in drug-resistant cells.2.The proliferation inhibition of cells in each group was observed by intervening cervical cancer resistant cells with cisplatin and the different concentrations of Polyphyllin Ⅰ.The expression levels of METTL3,CCND1 and MDR1 were determined by Western-blot and PCR fluorescence to investigate the mechanism of Polyphyllin Ⅰ to improve cisplatin sensitivity.3.Cervical cancer drug-resistant cells were cultured in vitro to build a cervical cancer nude mouse model of tumor xenograft,and the volume size,weight and growth of mouse tumors were recorded.Second,TUNEL staining was used to assess the effect of Polyphyllin Ⅰ on tumor growth in mice.The expression of relevant target gene proteins in tumor tissue was detected by immunohistochemical staining to further verify the mechanism of Polyphyllin Ⅰ resistance cell activity against cervical cancer described in the previous part.Results:1.Polyphyllin Ⅰ showed inhibitory effects on two different types of cervical cancer r esistant cells:HeLa/DDP cells and SiHa/DDP cells.And julifloor Polyphyllin Ⅰ promoted apoptosis in HeLa/DDP cells and SiHa/DDP cells.2.Polyphyllin Ⅰ is able to cause a large number of HeLa/DDP cells to arrest in G1 phase,promote their apoptosis,and can inhibit their proliferation,migration ability and clone formation ability.3.Polyphyllin Ⅰ can up-regulate Bax expression and downregulate Bcl-2 expression t hrough the P53/P21 pathway.4.The expression levels of METTL3,MDR1mRNA,and CCND1mRNA decreased i n cervical cancer HeLa/DDP cells after Polyphyllin Ⅰ and cisplatin intervention,and there was synergistic effect when Polyphyllin Ⅰ and cisplatin were combined,which could inc rease the sensitivity of HeLa/DDP cells to cisplatin.5.In the nude mouse xenograft model,Polyphyllin Ⅰ significantly inhibited the prolif eration of HeLa/DDP in cervical cancer,and the cell activity of tumor tissue of nude mi ce treated with Polyphyllin Ⅰ was statistically significant.In addition,P53 protein express ion was increased in the tumor tissues of nude mice,but the expression of CCNDl,MD R1 and METTL3 was significantly reduced.Conclusion:Polyphyllin Ⅰ inhibited the proliferation and migration ability and induced the apopto sis,and induced the apoptosis of the drug-resistant cells through the P53/P21 pathway.A t the same time,Polyphyllin Ⅰ and Cisplatin can reduce the expression levels of METTL 3mRNA,MDR1mRNA and CCND1mRNA in HeLa/DDP cells,and the combination has a synergistic effect.However,this study lays the foundation for more in-depth mechanisti c studies and has certain guiding significance.The study of the specific sensitivity mechan ism of Polyphyllin Ⅰ in cervical cancer resistant cells is not deep enough.This study laid the foundation for the subsequent exploration of the specific molecular mechanism of P olyphyllin I to enhance the sensitivity of cervical cancer resistant cells to cisplatin throug h MDR1mRNA and CCND1mRNA stability through METTL3,and has certain guiding si gnificance.