Molecular Mechanism Of Epithelial-Mesenchymal Transition And Radiation Resistance In Nasopharyngeal Carcinoma And The Intervention Effect Of Shengmai Yin

Background:Nasopharyngeal carcinoma(NPC)is a malignant squamous cell carcinoma caused by nasopharyngeal epithelium.Distant metastasis and local recurrence often lead to the failure of clinical treatment.More and more evidence shows that EMT is related to the production of tumor radiation resistance.It is of great significance to further clarify the mechanism of EMT and related genes in nasopharyngeal carcinoma and obtain the effect of Shengmai Yin in radiation resistance.Purpose:To construct the zebrafish transplanted tumor model of human nasopharyngeal carcinoma,clarify the molecular mechanism of radiation resistance of nasopharyngeal carcinoma cells,and explore the radiation resistance effect of Shengmai Yin in nasopharyngeal carcinoma.Methods:The transplanted tumor zebrafish model of human nasopharyngeal carcinoma radiation resistant strain CNE-2R was successfully constructed by microinjection technology.The inhibitory effect of Shengmai Yin on nasopharyngeal carcinoma radiation resistant cell line CNE-2R in zebrafish was detected by observing the fluorescence of the transplanted tumor.EMT morphological changes were observed in CNE-2R cells under microscope.RT-qPCR and Western Blot assays detected the expression of EMT markers in CNE-2R cells.Through database analysis and Geneclip3 text mining,LCN2,a gene highly related to radiation resistance and EMT of nasopharyngeal carcinoma,was obtained.RT-qPCR and Western Blot assays were used to detect the expression of LCN2 and EMT markers after Shengmai Yin on CNE2R.Clone formation assay,wound healing assay and Transwell invasion assay were used to detect the differences of cell growth,migration and invasion ability.The target of Shengmai Yin in the treatment of nasopharyngeal carcinoma was obtained by database mining,and then the protein interaction network and related pathway enrichment were constructed,verified by molecular docking and RT qPCR experiments.The key targets were discussed with EMT to obtain the mechanism of Shengmai Yin in the radiation resistance of nasopharyngeal carcinoma.Result:Zebrafish xenotransplantation model was successfully constructed.It was found that Shengmai Yin at the concentration of 5,10 and 20 μg/mL inhibited the proliferation and migration of nasopharyngeal carcinoma CNE-2R cell transplantation tumor.CNE-2R cells showed EMT and high expression of LCN2.Shengmai Yin can reduce the expression of LCN2 in CNE-2R cells and inhibit the process of EMT.Compared with CNE-2R group,the proliferation,migration and invasion of CNE-2R cells in Shengmai Yin high concentration administration group were weakened.65 active components and 445 targets of Shengmai Yin and 588 targets for NPC were screened.They had 66 common targets.1648 items of biological process,97 items of cell composition and 14 items of molecular function were obtained by GO functional enrichment;86 signal pathways were enriched to play a role by KEGG database.Molecular docking showed that the active compounds of Shengmai Yin had strong binding force with the targets.RT-qPCR results showed that compared with the control group,the expression multiples of AKT1,MAPK8,MAPK3,TNF and Jun in the high concentration group of Shengmai Yin were down regulated and related to EMT.Conclusion:Zebrafish xenotransplantation model can become an important model for NPC tumor research and drug evaluation.Shengmai Yin can inhibit the EMT process of CNE-2R cells,which may be related to its inhibition of LCN2 expression.And then it can reduce the proliferation,migration and invasion of tumor cells,and affect the radiation resistance of nasopharyngeal carcinomaThe mechanism of Shengmai Yin in the treatment of nasopharyngeal carcinoma involves multiple active components,action targets and key pathways related to EMT,which provides a scientific basis for the follow-up research and development of Shengmai Yin.