Differential Expression Study Of WNT7B,SFRP4 And RSPO2 Genes In The WNT Pathway

Objective In this experiment,the transcriptional expression levels of three genes,WNT7 B,SFRP4 and RSPO2,were examined in intact tissue specimens excised from patients with DD(Dupuytren’s Disease),and the protein expression networks constructed by the genes were explored and analyzed to elucidate the roles of the above genes in the pathogenesis of DD and whether they are related to each other.The study of the pathogenesis of DD will provide further theoretical basis for the realization of targeted treatment of the disease at the molecular level.Methods This study protocol was approved by the ethical review committee of the Third Affiliated Hospital of Inner Mongolia Medical University.The subjects selected for the study were specimens of diseased palmar tendon membrane tissues excised from three patients with DD who underwent surgical treatment in our hand,foot and ankle surgery from January 2022 to January 2023;the control group was selected from fragments of normal palmar tendon membrane tissues excised from three patients who underwent surgical treatment for carpal tunnel syndrome in our hospital at the same time.The tissues were immediately immersed in RNAwait preservation solution,placed in a 4℃ refrigerator overnight,and then removed and transferred to a-80℃ refrigerator for freezing.Uniform transcription sequencing analysis and detection: Firstly,total RNA was extracted from the tissues of DD experimental group and control group samples by Trizol method;Then m RNA was screened out from them for reverse transcription,and synthetic c DNA with length in 250-300 bp was screened,and finally NEB library was built for quality control;After the libraries were qualified for quality control,the Illumina platform was used for all libraries for Sequencing summary;according to the results,we performed differential significant gene analysis,gene function and pathway enrichment analysis.Results(1)RSPO2 was highly expressed in the experimental group,expression(FPKM)in the experimental group = 47.286,control group = 6.539,7-fold difference in expression,|log2Fold Change|=2.814>1,p=0.033<0.05.It is located on the negative strand of chromosome8 and annotated to be the No.2 encoded protein within the RSPO family.(2)SFRP4 was highly expressed in the experimental group,expression amount(FPKM)experimental group=8264.626,control group=2839.771,experimental group and control group|log2Fold Change|=1.541>1,p=0.053>0.05.Located on the negative strand of chromosome 7,annotated to belong to the SFRP family encoding protein number 4.(3)WNT7B was highly expressed in the experimental group,expression(FPKM)experimental group=1.702,control group=0,experimental group vs control group|log2Fold Change|=3.025>1,p=0.443>0.05.Located on the negative strand of chromosome 22,annotated as an encoded protein within the WNT pathway.Conclusion(1)A total of 2151 differentially significant genes were detected in lesional tissues of patients with palmar tendon contracture,with 504 up-regulated and 1647down-regulated genes expressed.(2)RSPO2 was highly expressed in the experimental group,and the KEGG enrichment results indicated that its binding to sulfur compounds within the Wnt signaling pathway regulated the development of connective tissue disease(DD);WNT7B and SFRP4 expression was up-regulated but not within the significant differential gene set.(3)WNT7B,RSPO2 and SFRP4 are all located upstream of the WNT signaling cascade and are coding proteins.