ZMIZ1 Affects Invasion And Metastasis Of Tongue Squamous Cell Carcinoma By Regulating Notch Signaling Pathway

Background and Objective:Tongue squamous cell carcinoma(TSCC)is the highest squamous cell carcinoma with oral and maxillofacial morbidity and mortality.The 5-year survival rate is only 32-54%.At present,comprehensive treatment is the main treatment.Because of the special TSCC,the complications of comprehensive treatment and the poor quality of life of patients,gene-targeted therapy has become a research hotspot in the treatment of TSCC in recent years.Our previous study showed that evodiamine(EVO)inhibited the proliferation,metastasis and apoptosis of TSCC cells,but the specific mechanism was not clear.In this study,bioinformatics analysis was performed by whole genome sequencing and experiments to find potential biomarkers of TSCC and provide new ideas for gene-targeted therapy.Methods:1.Identification of differential gene expression after treatment of TSCC cells with EVO by Affymetrix gene expression profiling chip and ingenuity pathway analysis(IPA).2.The lentivirus LV-ZMIZ1-RNAi was transfected into tongue squamous cell carcinoma CAL-27 cells,and the expression of ZMIZ1 was knocked down.The effects of ZMIZ1 on proliferation,migration and apoptosis in the development of TSCC were studied by CCK-8,wound healing test and flow cytometry.Then,the mechanism was explored by qRT-PCR and Western Blot.3.Establish a lung metastasis model of nude mice through the blood.The CAL-27 cells transfected with lentivirus LV-ZMIZ1-RNAi and LV-Ctrl-RNAi were injected into the corresponding groups of nude mice through the tail vein,and taken after 50 days.The lungs were subjected to HE pathological staining,and the metastases were counted and analyzed.The mechanism was explored by immunohistochemistry,qRTPCR and Western Blot.Results:1.Microarray results showed that there were 1,243 differentially expressed genes following treatment with CAL-27 cells;684 genes were up-regulated and 559 were down-regulated.Classical pathway analysis revealed a total of 89 signal transduction pathways with up-regulated gene set enrichment and A total of 39 signal transduction pathways were enriched for the down-regulated genes.Further studies revealed that the upstream gene ZMIZ1 after Evodiamine treatment was significantly inhibited,and the Notch signaling pathway was also significantly inhibited.2.In vitro experiments showed that LV-ZMIZ1-RNAi transfected CAL-27 cells could inhibit cell proliferation,migration and promote cell apoptosis compared with the control group(p<0.05).The results of qRT-PCR and Western Blot showed that knockdown of ZMIZ1 could reduce the expression of Notch1,Jagged1,MKP-1,SSBP2 and MMP7 in CAL-27 cells(p<0.05).3.Through the tail vein lung metastasis model,the number of metastatic nodules formed in the lungs of nude mice injected with lentiviral LV-ZMIZ1-RNAi was lower than that of the control nude mice(p<0.05).And the expression of Notch1,Jagged1,MKP-1,SSBP2 and MMP7 was lower(p<0.05).Conclusion: Both in vitro and in vivo experiments indicated that knockdown of ZMIZ1 gene inhibits invasion and metastasis of tongue squamous cell carcinoma by regulating Notch signaling pathway.It is suggested that ZMIZ1 may be a potential target for the treatment of tongue squamous cell carcinoma.