Antitumor Effect Of Salinomincy Combined With Hydroxychloroquine On Colorectal Cancer Cells

Background and PurposesColorectal cancer is one of the malignant tumors endangering public health in China,and its incidence rate and mortality rate are high.The common treatment methods for colorectal cancer include surgery,radiotherapy and chemotherapy.Due to the emergence of drug resistance,most chemotherapy can not achieve the desired effect in the late stage of treatment^[1].Therefore,it is necessary to find a reasonable combination of drugs to improve the efficacy of chemotherapy.The abnormal activation of Wnt/β-Catenin signaling pathway is an important inducement of tumorigenesis.In Colorectal cancer,Wnt pathway has a high activation rate of more than 80%.Therefore,inhibiting Wnt pathway has also become a potential direction to treate tumor.Salinomycin has been proved to target tumor stem cells,and also has a certain inhibitory effect on Wnt signaling pathway.It can also specifically induce or inhibit autophagy in different tumor cells.In recent years,in addition to the burgeoning research and development of new drugs,new uses of old drugs have also been advocated,including exploring the new functions and new uses of mature drug preparations.Therefore,Salinomycin is expected to be used in the anti-tumor treatment of clinical patients in the future.Autophagy is ubiquitous in eukaryotic cells,it can provide energy for cell activities by degrading substances such as organelles.Some studies have shown that inhibition of autophagy can improve the effect of radiotherapy,and the combination of autophagy inhibitors and chemotherapy drugs can also sensitize the anti-cancer effect of this chemotherapy drug.Hydroxychloroquine is an autophagic inhibitor that acts in the late stage.Because of its good safety,more than 20 clinical trials of Hydroxychloroquine combined with radiotherapy and Hydroxychloroquine combined with chemotherapy are currently under way.The combination of radiotherapy,chemotherapy and Hydroxychloroquine is expected to provide new ideas and methods for the treatment of tumors.In order to explore the anti-tumor effect of Salinomycin combined with Hydroxychloroquine on Colorectal cancer cells,We analyzed the effects of Salinomycin,Hydroxychloroquine alone and their combination on cell activity in this study,such as proliferation,autophagy,apoptosis,etc.The purpose was to determine whether the combination of Salinomycin and Hydroxychloroquine has a synergistic effect on the treatment of cancer and continue to explore the relevant mechanisms.It aimed to provide new theoretical basis for clinical diagnosis and treatment of Colorectal cancer patients.MethodsHuman Colorectal cancer cell line HCT116 was mainly used as the research object1.CCK-8 experiment and colony formation experiment were used to explore the effects of Salinomycin,Hydroxychloroquine alone and combination therapy on cell proliferation.2.q PCR and Western Blot experiments were used to explore the effect of Salinomycin on the expression of downstream genes and proteins of Wnt pathway,so as to verify the effect of Salinomycin on Wnt pathway.3.Flow cytometry was used to detect the effect of combination therapy on cell cycle distribution and apoptosis of different treatment groups.4.The effects of Salinomycin alone and Salinomycin combined with Hydroxychloroquine on autophagy were detected by acridine orange staining and Western blot.5.Human Colorectal cancer cell SW480 was treated with Salinomycin,hydroxychloroquine alone or in combination,and the combination therapy of two drugs on proliferation was verified by CCK-8 and colony formation experiments.6.HCT116 cells and SW480 cells were treated with Salinomycin and 3-MA alone or in combination,in order to observed cell proliferation and explore whether the synergistic effect of Salinomycin and Hydroxychloroquine was related to autophagy.Conclusion1.Salinomycin inhibited the proliferation of HCT116 by inhibiting the Wnt signal pathway.Hydroxychloroquine also inhibited cell proliferation to a certain extent.2.The combination of Salinomycin and Hydroxychloroquine inhibited the proliferation of Colorectal cancer cells.3.Combined application of Salinomycin and Hydroxychloroquine blocked the cell cycle of HCT116 at G0/G1 phase.4.The combination of drugs promoted cell apoptosis.5.Salinomycin induced autophagy of HCT116 cells,which was inhibited by Hydroxychloroquine.6.Combined application of Salinomycin and Hydroxychloroquine significantly inhibited the cell proliferation of SW480.7.The inhibitory effect of the combination of Salinomycin and Hydroxychloroquine on cell proliferation was simulated by the combination of Salinomycin and 3-MA.Results1.Salinomycin inhibited the proliferation of cancer cells and induced apoptosis by inhibiting the Wnt signal pathway,Salinomycin has a clear anti-tumor effect.2.Salinomycin induced protective autophagy in cancer cells,which can be inhibited by Hydroxychloroquine.3.Hydroxychloroquine inhibited autophagy while cooperating with Salinomycin to block cell cycle at G0/G1 phase,inhibited cell proliferation and activated cell apoptosis through mitochondrial pathway.Thus,the anti-tumor effect of Salinomycin was enhanced.Salinomycin combined with Hydroxychloroquine can synergistically inhibit tumor growth.