Study The Pathogenesis Of Mycosis Fungoides Through WGCNA

Part Ⅰ:Identifies novel biomarkers for mycosis fungoides using WGCNAObjective:Mycosis fungoides is the most common form of cutaneous T-cell lymphoma and represents a complex series of diseases with various manifestations.Patients with mycosis fungoides(MF)developing tumors or extracutaneous lesions usually have a poor prognosis with no cure has so far been available.To identifythe pathogenesis and potential novel biomarkers for MF,a genomic mapping of 41 cutaneous lymphoma biopsies was used to explore for significant genes.Methods:The gene expression profiling datasets of MF were obtained from Gene Expression Omnibus database(GEO).Gene modules were simulated using Weighted Gene Co-expression Network Analysis(WGCNA)and the top soft-connected genes(hub genes)was filtrated with a threshold(0.5).Subsequently,module eigengenes were calculated and significant biological pathways were enriched based on the KEGG database.Results:Four genetic modules were simulated with 3,263 genes collected from the whole genomic profile based on cutoff values.Significant diseases genetic terminologies associated with MF were found in black module.Subsequently,13 hub genes including CFLAR,GCNT2,IFNG,IL17A,IL22,MIP,PLCGI,PTH,PTPN6,REGIA,SNAP25,SUPT7L and TP63 were shown to be related to cutaneous T-cell lymphoma(CTCL)and Adult T-cell Lymphoma/Leukemia(ATLL).Conclusion:In summary,in addition to the reported genes(IL17F,PLCG1,IFNG,PTH)in CTCL/ATLL,the other high instable genes may serve as novel biomarkers for the regulation of the biological processes and molecular mechanisms of CTLT(MF/SS)Part Ⅱ:Expression and significance of hub genes in CTCL Myla and Hut-78 cell lines in vitroObjective:To explore the expression and significance of hub genes in CTCL Myla and Hut-78 cell lines in vitro.Methods:RT-qPCR was used to analyze the expressions of hub genes IL17A,PLCG1,CFLAR,GCNT2,REGIA,PTPN6,SNAP25 and STAT3 in CTCL Myla and Hut-78 cell lines.The peripheral T lymph cells from healthy volunteers were taken as control group.An immunohistochemical study was performed to measure the expression of IL17A and STAT3 in Myla cell line.Results:The RT-qPCR results of Myla and Hut-78 cell lines showed significantly up-regulated PLCG1,IL-17A and STAT3,compared with peripheral T lymph cells from healthy volunteers.CFLAR,GCNT2,PTPN6 were significantly down-regulated.SNAP25 was significantly up-regulated in Hut-78 cell line,REGIA was significantly up-regulatedin Myla cell line.The expression of IL-17A and Stat3 were significantly higher in Myla cell line.Conclusion:The expressions of the Hub genes IL-17A,GCNT2,REGIA and SNAP25 in the MF cell lines were consistent with the results of WGCNA.IL-17A and GCNT2 were also significantly different in Hut-78 cell line which indicates that it may be related to the genesis of mycosis fungoides.Part Ⅲ:Expression and significance of hub genes IL17A in tissues of patients with mycosis fungoidesObjective:To explore the expression and significance of hub gene IL17A in tissues of patients with mycosis fungoides.Methods:An immunohistochemical study was performed to measure the expression of IL-17A,IL-17F,IL-22 and IL-23 in the lesional specimens from 31 patients with mycosis fungoides and skin specimens from 20 controls with psoriasis.Results:The expressions of IL-17A,IL-17F,IL-22 and IL-23 were significantly higher in non-tumor state MF and tumor stage MF tissues than in psoriasis tissues(P<0.05).The expressions of IL-17A IL-17F,IL-22 and IL-23 were significantly lower in tumor stage mycosis fungoides tissues than innon-tumor stage MF tissues.(P<0.05).Conclusion:As MF progressing,the expressions ofIL-17A,IL-17F,IL-22 and IL-23 proteins were lower in the MF lesions.The hub gene IL-17A and its related cytokines were associated with mycosis fungoides progression.But whether it was tumor suppressor gene needed further researchs.