Study On The Roles Of Inlfammatory Factors And Micrornas In The Molecular Pathogenesis Of Gastric Malt Lymphoma

Objective:MicroRNA is a class of endogenous non-protein-coding RNA about19-25nucleotides inlength, binding to the3’UTR of target mRNA through base pairing principles, and thusde-gradates target mRNA or inhibits translation of target mRNA, thus plays a biologicaleffect. MicroRNA plays a dual effect of tumor suppressor genes or oncogenes involved intumor metabolism, such as cell proliferation and apoptosis, tumor invasion, it also plays avital role in the development of tumors, but different tumors have different microRNAexpression profile. Gastric MALT lymphoma accounts for40%-60%of the MALTlymphoma. Some studies showed that MALT lymphoma was based of gastric mucosalymphocyte proliferation of HP infection, anti-HP therapy achieved clinical completeremission in part of MALT lymphoma. The role of microRNA is not yet entirely clear inthe progress of chronic inflammation to tumor transformation. In this study, microRNAexpression changes in gastric MALT lymphoma and pathogenesis were studiedpreliminary,providing the theoretical basis and experimental data for the prevention ofMALT lymphoma, and targeted therapy.Methods:(1)Detection of microRNA expression profile differences in gastric MALT lymphomatumor tissue and peritumoral organization.①Cases of gastric MALT lymphoma were collected from Changhai Hospital, accordingto the HE form, immune mark combined WHO2008lymphoid hematological neoplasmdiagnostic criteria to determine the gastric MALT lymphoma.②microarray chip todetect microRNA expression profile changes, then to extract total RNA from tumors andtumor adjacent tissues, and microRNA microarray to screen and analysis data.③thereal-time quantitative RT-PCR to detect14cases of gastric MALT lymphoma tumortissue and peritumoral tissue gene: microRNA-188-5p, microRNA-134, microRNA-150,microRNA-100, microRNA-378, microRNA-155gene expression differences. UsingSYBR Green Realtime PCR Master Mix real-time quantitative PCR to amplificate, U6asinternal control, three wells.2-△Ct to calculate the six genes of the tumor and tumoradjacent tissues and the relative expression levels.④According to clinical data,SPSS13.0software package to analyze the relationship between microRNA and theprognosis of patients.(2)Detection the target gene protein of microRNA differentially expressed genes ①(http://www.mirbase.org/and http://www. Targetscan.org/) to detect the target genesof microRNA-100and microRNA-378, they are TNFAIP8, SMARCA5and MMP14,MAPK1.②selected50cases of gastric MALT lymphoma patients undergoing tumor andadjacent to the tumor histopathology, the LSAB method to detect the expression ofMAPK1, TNFAIP8, SMARCA5, MMP14.Results(1) Detection of microRNA expression profile differences between in gastric MALTlymphoma tumor tissue and peritumoral organization①MicroRNAs microarray analysis results: the expression difference which was morethan twice was significant, up-regulated expression genes in tumor tissue: microRNA-15a,microRNA-155, microRNA-16, microRNA-100, microRNA-142-3P, microRNA-150.Down-regulated genes: microRNA-375, microRNA-188-5p, microRNA-663,microRNA-134, has-microRNA-31, microRNA-1246, microRNA-200c, microRNA-12245p, microRNA-378, micoRNA-200b.②Analysis of RT-PCR results: microRNA-188-5p of tumor VS peritumoral:8.52±0.91VS9.05±1.03, P>0.05; microRNA-100of tumor VS peritumoral:6.83±1.18VS6.92±1.36, P>0.05; microRNA-150of tumor VS peritumoral:4.78±1.37VS5.32±1.34,P>0.05; microRNA-378of tumor VS peritumoral:9.13±1.04VS7.38±1.44, P<0.05;microRNA-134of tumor VS peritumoral:12.92±0.05VS12.46±1.06, P>0.05;microRNA-155of tumor VS peritumoral:8.52±0.92VS9.05±1.03. P>0.05.③Analysis using SPSS software package, chi-square test of the clinical data ofmicroRNA-378with or without gastric ulcer P=0.01. Of miRNA-100HP-positive, P=0.05. Of miRNA-155with or without violation of myometrial P=0.03.(2) Results of target gene protein test of microRNA differentially expressed genes Targetgene protein MAPK1TNFAIP8, MMP14of miRNA-378and microRNA-100were lowlyexpressed in tumor tissue using immunohistochemical detection, SMARCA5showed ahigh expression.Conclusion:(1) MicroRNA expression was different between tumor tissue and tumor adjacent tissuesin gastric MALT lymphoma, of which16genes were significantly differentiallyexpressed.(2) In gastric MALT lymphoma, the microRNA-378expression was higher in tumortissue than in the peritumoral organizations, the difference had statistical significance; MicroRNA-378expression was related to the occurrence of gastric ulcer; HP infectionmay increase the expression of microRNA-100, leading to lymphoma; microRNA-155was related to tumor aggressiveness of patients.(3) MicroRNA-378gene that caused stomach ulcers may be through the target geneMMP14, MAPK1protein expression, HP infection may increase the expression ofmicroRNA-100gene, further activate TNFAIP8, SMARCA5gene expression, resultingin tumorigenesis.