Evodiamine Inhibits The Proliferation Of Wilms Tumor And Induces Apoptosis In Vivo

Evodiamine（EVO）,a natural compound and quinolones bioactive alkaloid,is extracted from the dried fruits of Evodia rutaecarpa.Recently,EVO is widely used in cancer research.EVO has inhibitory effects on various tumors including colon cancer and ovarian cancer;its mechanism may be related to inhibiting proliferation and promoting apoptosis.Although EVO has a prominent effect in tumor development,it hadn’t yet used in clinical practice.This study evaluates controversial issues with conflicting conclusions through systematic reviews and meta-analysis,and provides evidence support for clinical translation.Numerous studies have shown that EVO has inhibitory effects on a variety of tumors.Preliminary studies of our group have shown that EVO has an inhibitory effect on the growth of tongue squamous cell carcinoma,but there is no related research on the inhibition of Wilms tumor by EVO.In this study,animal experiments were conducted to explore the inhibitory effect of evodiamine on Wilms tumor,and to preliminarily explore the relevant mechanism to provide evidence for clinical transformation.Part I Synergistic Antitumor Effects of Evodiamine in Animal Experiments: A Systematic Review and Meta-AnalysisObjective: Evodiamine has made remarkable progress in tumor research,but there are still controversial conclusions,and it has not been used in clinical treatment.This subject evaluates the synergistic antitumor effect of evodiamine in vivo experiments through systematic review and meta-analysis,and provides a basis for clinical translation.Methods: Literature databases were searched,including Pub Med,Scopus,Sino Med and China Knowledge Resource Integrated Database（CNKI）.Literature screening was performed according to the established inclusion and exclusion criteria.Data was extracted from the selected literature.In this study,tumor volume and tumor weight were used as outcome indicators,and Stata 15.0 software was used to analyze and detect relevant data.The heterogeneity was explored,and if there was heterogeneity,it was further explored through regression analysis.Publication bias was assessed by Begg’s test,and stability of results was assessed by sensitivity analysis.Results: A total of 7 articles were included in this study,including 7 articles with tumor volume and 4 articles with tumor weight.There was a statistically significant difference in tumor volume between the combination treatment group and the control group（SMD=-25.64,95% CI:-5.77,-3.13;I2 = 92.5%,P<0.05）.There was a statistically significant difference in tumor weight between the EVO group and the control group（SMD=-8.91,95% CI:-16.37 to-1.44;I2 = 94.1%,P<0.05）.Conclusion: This study shows that evodiamine has synergistic antitumor effects in vivo compared with monotherapy,and may be a potential chemotherapeutic drug for tumor treatment.This will provide new ideas for tumor treatment and clinical translation.Part II: Inhibitory Effects of Evodiamine on the Proliferation and Induction of Apoptosis of Wilms TumorObjective: To establish a nude mouse model of subcutaneously transplanted tumor,to investigate the inhibitory effect of evodiamine on Wilms tumor,and to observe the proliferation and apoptosis-related markers,so as to provide new evidence for the clinical transformation of evodiamine.Methods: The subcutaneous transplanted tumor model was established by subcutaneous injection of Wilms tumor cells,and the inhibitory effect on tumor was observed by administration of evodiamine.The heart,liver,spleen,lung and kidney of nude mice were stained with HE,and the tumor tissue was stained with HE.The proliferation and apoptosis-related proteins were detected by immunohistochemical staining,and the tumor tissue was observed by transmission electron microscope to detect the apoptosis of tumor cells.Results: Nude mice subcutaneously transplanted tumor model was successfully established.Compared with the control group,the tumor volume of the evodiamine treatment group was significantly reduced（P<0.05）,and the EVO 20mg/kg group had a more significant inhibitory effect on the tumor than the 10mg/kg group（P<0.05）.Observation under HE staining microscope showed that compared with the control group,the nucleocytoplasmic ratio in the tumor tissue of the experimental group decreased,and the mitotic figures of the cells decreased.The expression of-67 was down-regulated,and the expression of Caspase-3 and Caspase-9 was up-regulated;the results of transmission electron microscopy showed that the EVO 10mg/kg group had early cell apoptosis,and the 20mg/kg group had late cell apoptosis;TUNEL staining results showed that the tumor in the experimental group was Compared with the control group,the positive expression in the tumor tissue increased;under the HE staining microscope,the heart,liver,spleen,lung and kidney tissues of the experimental group had no obvious abnormality;There was no statistical difference in the control group,and there was no significant difference in body weight among the three groups.Conclusion: Evodiamine inhibits the development of Wilms tumor in vivo.After administration,the proliferation-related protein Ki-67 is down-regulated,and the apoptosis-related proteins caspase-3 and caspase-9 are up-regulated.