Effects Of Doramectin On Apoptosis Of Esophageal Cancer Cells

Characterized by high morbidity and mortality,esophageal cancer ranks among the top ten in the world of cancer and has become one of the malignant diseases affecting human health worldwide.Currently,according to the constitution and degree of disease of patients with esophageal cancer,the treatment methods mainly include surgic al resection of the lesion site,concurrent chemoradiotherapy,surgery combined with neoadjuvant chemotherapy,biological therapy,targeted therapy,etc.,but the therapeutic effect is not good,so the research and development of therapeutic drugs for esophageal cancer is crucial.Studies have found that macrolides play a key role in reversing multidrug resistance of tumor cells,and these drugs have an inhibitory effect on the growth of tumor cells to a certain extent.Besides,these drugs can also induce apoptosis and autophagy of cells,but specific theoretical basis studies are scarce.Therefore,Doramectin(DOR),a macrolide antibiotic family,was selected in this study to explore whether the drug could further inhibit the proliferation of esophageal ca ncer cells by inducing apoptosis.In this study,MTT experiment and scratch experiment were used to explore whether the proliferation activity and migration ability of DOR treated cells were changed.The main morphological changes of cells treated by DOR were observed by an inverted microscope and transmission electron microscope respectively.Flow cytometry detected the changes of apoptosis rate and DNA content at different periods after DOR treatment.Western blotting detected the specific changes of apoptotic protein expression in cells treated with DOR at different concentrations.The animal model was established,and the effect of DOR on the proliferation and apoptosis of esophageal cancer cells was deeply discussed through experiments such as HE,TUNEL,and immunohistochemical staining,to provide sufficient theoretical basis for the search for new adjuvant drugs for the clinical treatment of esophageal cancer.Results are as follows:(1)Eca109 and EC9706 cells were treated with DOR at different concen trations,and the results showed that the proliferation activity and migration ability of both cells showed a downward trend with the gradual increase of drug dose.(2)Under an inverted microscope,Eca109 and EC9706 cells were observed to become round,shrink,and gradually sparse with a decreasing number after treatment.Under the transmission electron microscope,it was found that the cell volume decreased,the nucleolus shrank,and apoptotic corpuscles appeared.(3)Flow cytometry was used to detect the changes in DNA content of Eca109 cells at different stages after DOR treatment.The results showed that the DNA content of THE CELLS at the G2/M stage increased significantly with the increase of drug dose(P < 0.001).(4)Flow cytometry detected the apoptosis of cells treated with DOR at different concentrations.The results showed that the apoptosis rate increased significantly with the increase of drug dose(P < 0.0001).The expression of apoptotic protein in the cells was detected by Western blotting experiment.The results showed that the expression level of Bcl-2 protein was decreased while that of Bax protein was increased.Besides,cytochrome-c,Caspase-3,and Caspase-9 protein were all increased.(5)The DOR inhibited the growth of Eca109 cells in the in vivo tumor-bearing nude mice model,but there was no significant difference in body weight(P < 0.05),the tumor volume and wet weight of the treated group decreased significantly,and the difference was significant(P <0.01);TUNEL staining showed that the number of apoptotic cells in tumor tissues increased significantly after drug treatment.Immunohistochemical results showed that the positive expressions of caspase-3 and Caspase-9 were significantly increased in the drug-treated group.In conclusion,DOR can inhibit the proliferation and migration of esophageal cancer cells and block the cell cycle at the G2/M stage.Mitochondrial pathway and caspase pathway are involved in the whole process of apoptosis.Therefore,the theoretical basis of DOR induced apoptosis of esophageal cancer cells is of great significance for the clinical treatment of esophageal cancer.