SPINK5 To Regulate The Notch Signaling Pathway By Targeting TFG Controls EMT Of Oral Squamous Cell Carcinoma

Objective: Oral squamous cell carcinoma(OSCC)accounts for more than 90% of all oral malignant tumors.Differences in global prevalence can be attributed to differences in exposure to carcinogenic risk factors,with a 5-year survival rate of 50%.The prognosis is extremely poor,and there is a lack of effective treatment methods.Therefore,this study aims to screen OSCC diagnosis and treatment targets through non-invasive saliva samples,providing a theoretical basis for the clinical development of effective targeted treatment plans and improved prognosis.Methods: 1)Differential expression protein screening was performed on 9 cases of OSCC and 3 cases of control fresh tissue using tandem mass spectrometry(TMT)proteomics technology,and candidate markers were identified using various bioinformatics analysis platforms.2)Differential metabolite screening was performed on 9 cases of OSCC and 3 cases of control fresh tissue using non-targeted metabolomics technology,and candidate markers were identified using various bioinformatics analysis platforms.3)A diagnostic model was established using multi-omics integration technology to identify potential target genes;4)SPINK5overexpression and silencing stable strains were established to study the effects of SPINK5 on the cell biology of OSCC,and the possible regulatory mechanism of SPINK5 was initially explored from cytology;by establishing a nude mouse tumorigenesis model,the tumorigenic effect of SPINK5 in vivo was analyzed;immunoprecipitation-mass spectrometry technology was used to analyze the possible downstream regulatory mechanism of SPINK5,providing research ideas for further studying the mechanism of SPINK5.Results: 1)Through proteomics research,679 differentially expressed proteins were screened out,of which 274 were up-regulated and 405 were down-regulated;2)Through non-target metabolomics research,a total of 225 differentially expressed metabolites were identified in cation mode,including 194 down-regulated metabolites and 31 up-regulated metabolites;a total of 30 differentially expressed metabolites were identified in anion mode,including 28 down-regulated metabolites and 2up-regulated metabolites.3)Using proteomics data,metabolomics data,and TCGA transcriptomics dataset,8 core proteins related to OSCC were screened out through bioinformatics analysis: LGALS1,CSTA,SPINK5,PTX3,ENDOU,ASAH1,RAB11FIP1,and ST6GALNAC1;4)clinical sample test results showed that SPINK5 is lowly expressed in tumors,and cellular studies have confirmed that SPINK5 has important tumor-suppressing effects such as inhibiting cancer cell proliferation,promoting apoptosis,and inhibiting migration,which was verified through nude mouse tumorigenesis experiments;its possible mechanism of action includes regulating epithelial-mesenchymal transition(EMT)and NOTCH1-related signaling pathway regulation;additionally,SPINK5-interacting upstream and downstream proteins were screened using immunoprecipitation combined with mass spectrometry,providing research directions for further studying the regulatory mechanism of SPINK5 in OSCC.Conclusion: Proteomics,metabolomics,and transcriptomics are effective technical means for screening tumor markers,and combining multi-omics methods can effectively identify target;SPINK5 is an important OSCC-related tumor suppressor gene and may be a potential target for the diagnosis and treatment of OSCC.