Evaluation Of The Therapeutic Effect Of Cisplatin Combined With Metformin On Endometrial Cancer At Different Concentrations Of Glucose In Vitro

Purpose: Obesity and hyperglycemia are not only closely related to the occurrence and development of type I endometrial cancer,but also to the high mortality rate of endometrial cancer.Obesity is associated with poor chemotherapy in patients with endometrial cancer with abnormal glucose metabolism.Endometrial cancer cells under high glucose promote the growth and proliferation of endometrial cancer cells by regulating the AMPK/mTOR/S6 signaling pathway.Metformin acts on the AMPK target to activate the AMPK signaling pathway,thereby exerting an anti-tumor effect.In addition,metformin can improve the anti-tumor ability of the chemotherapeutic drug cisplatin,and metformin combined with cisplatin has synergistic anti-tumor effect compared with cisplatin alone.Therefore,in this study,we applied established endometrial cancer cell lines(HEC-1-B,Ishikawa)to investigate the anti-tumor mechanism of metformin,cisplatin,and metformin combined with cisplatin in different glucose concentrations.Methods: Endometrial cancer cell lines HEC-1-B and Ishikawa were cultured in vitro with different sugar concentrations(low sugar: 5.5mmol/L,high glucose: 25.5mmol/L),with different concentrations of metformin,cisplatin and metformin combined with cisplatin in the two cells.The CCK-8 method was used to evaluate the effects of the above three drugs on the proliferation inhibition of endometrial cancer cells at different sugar concentrations.The PI staining method and the Annexin V/PI method were used to detect metformin,cisplatin and metformin combined with cisplatin on the cycle and apoptosis of two endometrial cancer cells at different sugar concentrations.The changes of AMPK/mTOR/S6 signaling pathway related proteins were detected by Western protein imprinting experiment,and the changes of cell cycle protein cyclinD1 and cell anti-apoptosis protein MCL-1 were detected by using the above drug intervention in different sugar concentration environment.To clarify the effect of metformin,cisplatin and metformin combined with cisplatin on the growth and metabolism of endometrial cancer cells in high sugar environment and its mechanism.Results: Metformin,cisplatin and metformin combined with cisplatin inhibited the proliferation of HEC-1-B and Ishikawa cells in a dose-dependent manner at different sugar concentrations.Metformin,cisplatin,and the combination of the two inhibited proliferation of Ishikawa and HEC-1-B cells by G0/G1 phase,G2/M phase,and S phase,respectively.Western blotting results showed that with the increase of drug concentration,the expression of cyclinD1,MCL-1 and p-S6 in HEC-1-B and Ishikawa cells decreased,and the expression of p-AMPK was enhanced.In addition,when the drug concentration was the same,the high glucose concentration up-regulated p-AMPK and down-regulated cyclinD1,MCL-1,and p-S6 were significantly lower than that of low sugar.Conclusion: Metformin,cisplatin,and metformin combined with cisplatin inhibited the proliferation inhibition of endometrial cancer cells,and the combination of metformin and cisplatin increased cisplatin-induced AMPK activation,inhibited the expression of AMPK/mTOR/S6 signaling pathway,and blocked The S phase of cell cycle and induction of apoptosis enhance the anti-tumor effect of cisplatin.At high glucose concentrations,metformin,cisplatin,and metformin combined with cisplatin inhibited the proliferation of HEC-1-B and Ishikawa cells less than low glucose.By using AMPK agonist metformin to lower blood glucose while targeting the above signaling pathways,combined with the chemotherapy drug cisplatin to produce synergistic effects,improve the efficacy of chemotherapy,and provide new ideas for individualized treatment of patients with abnormal glucose metabolism.