To Study The MicroRNA Expression Profilesinfor AIDS Related Diffuse Large B-cell Lymphoma

Background:The essence of acquired immune deficiency syndrome related lymphoma(ARL) is malignant transformation of immune cells. Immune deficiency is the pathologic basis of both ARL and acquired immunodeficiency syndrome(AIDS). ARL is one of the most common AIDS related malignancies. AIDS related diffuse large B-cell lymphoma(AIDS-DLBCL) is a group of heterogeneous tumors associated with HIV infection. Lesions occurred in the lymph nodes and extranodal organs, and the tumors were aggressive with a poor prognosis. The pathogenesis of ARL has not been clearly elucidated. Micro RNAs(mi RNAs), which negatively regulate gene expression at the post-transcriptional level by degradating target m RNA or repressing protein translation, are a class of endogenous non-coding RNAs with the length of 18-23 nucleotides. Mi RNAs may function as oncogenes or tumor suppressors in tumor initiation and progression, so they can be used as markers for tumor diagnosis and prognosis and potential targets for therapy. In recent years, the pathogenesis,diagnosis, prognosis and treatment of AIDS-DLBCL were poorly understood due to the limited researches.The purpose of the study:1. To screen the differently expressed mi RNAs in AIDS-DLBCL and non-AIDS-DLBCL tissues by applying Exiqon mi RCURYTMLNA mi RNA microarray detection and establish the reliable mi RNA expression profile of AIDS-DLBCL.2. To predict the target genes of abnormally expressed mi RNAs in AIDS-DLBCL tissues and reveal the regulatory relationship between mi RNAs and their target genes.3. To provide a basis for the study of the pathogenesis, diagnosis and therapy oflymphoma.Materials and methods:1. The establishment of mi RNA expression profile in AIDS-DLBCLWe chose 6 cases of AIDS-DLBCL tissues diagnosed by several hospitals in Yunnan Province from December in 2010 to February in 2013 as experimental group. 4 cases of non-AIDS-DLBCL tissues diagnosed by Affiliated Hospital of Dali University during the same period were selected as control samples. All the samples were paraffin-embedded tissues and detected by HE staining. The total RNA was extrated from samples with Trizol reagent according to the manufacturer’s instructions. After assessing the quality of RNA by spectrophotometry and agarose gel electrophoresis,mi RNA was labeled with fluorescence and hybridized with the mi RNA microarray chip. Subsequently, the chip was scanned and analyzed to obtain the mi RNA expression profile in AIDS-DLBCL.2. Prediction of the target genes of differently expressed mi RNAs in AIDS-DLBCL The target genes of differently expressed mi RNAs in AIDS-DLBCL were predicted by bioinformatic softwares(Mi Randa, mi RBase and Target Scan). The biological functions of the target genes were further analyzed using Gene Ontology(GO) and Kyoto Encyclopedia of Genes and Genomes(KEGG) databases Results The establishment of mi RNA expression profile in AIDS-DLBCL1. Both the concentration and quality of total RNA extracted from 6 cases of AIDS-DLBCL and 4 cases of non-AIDS-DLBCL tissues were ideal. The A260/A280 values ranged from 1.8 to 2.1 and the A260/A230 values were over 1.8. The agarose gel electrophoresis showed that the bands of 28 S and 18 S r RNA were clear without contamination of DNA and protein.2. The scatter diagram showed the correlation of hybridization signal between samples in tumor group or control group was high, reflecting the biological properties of samples in th same group were similar. However, the correlation coefficient(R) of two groups was 0.98 due to the expression of a few of mi RNAs was changed.3. Differently expressed mi RNAs exsit in AIDS-DLCBL and non-AIDS-DLBCLtissues. 64 mi RNAs were up-regulated and 44 mi RNAs were down-regulated in AIDS-DLBCL. Analysis by T-test showed that 23 mi RNAs were differently expressed in AIDS-DLBCL tissues, including 6 up-regulated mi RNAs(ebv-mi R-BART8-5p、mi R-185-3p、mi R-1469、mi R-744-5p、mi R-4505、mi R-4497) and 17 down-regulated mi RNAs(mi R-664a-5p、mi R-502-3p、mi R-660-5p、mi R-96-5p、mi R-29a-5p、mi R-182-5p、mi R-769-5p、mi R-129-2-3p、mi R-532-5p、mi R-664b-5p、mi R-320a、mi R-891a-5p、mi R-30b-5p、mi R-30d-5p、mi R-3607-3p、mi R-3653、mi R-32-3p).Prediction of the target genes of differently expressed mi RNAs in AIDS-DLBCLThe prediction of the target genes of the 23 mi RNAs was performed using three kinds of softwares, including Mi Randa, mi RBase and Target Scan. In order to reduce the false-positive rate, we chose the intersection of genes predicted by at least 2 kinds of softwares as the target genes of mi RNAs. 4 genes were predicted to be associated with the initiation and progression of AIDS-DLBCL, that were ZAP70, TNFRSF13 C,IGGL1(target genes of mi R-185-3p) and CD79A(target gene of mi R-744-5p). These results also suggested that mi R-185-3p and mi R-744-5p, involved in regulation of HIV infection and the progression of AIDS-DLBCL, might become the molecular markers of AIDS-DLBCL.Conclusion:This study obtained several crucial mi RNAs and their target genes in AIDS-DLBCL. We concluded that the up-regulation of mi R-185-3p and mi R-744-5p reduced the expression of ZAP70, TNFRSF13 C, IGLL1 and CD79 A, resulting in immunodeficiency and increasing the susceptibility to HIV infection and the incidence of AIDS-DLBCL. This study provided a foundation for further research on the biological effects of the differently expressed mi RNAs and their target genes in AIDS-DLBCL.