Study Of Molecular Genetics On Hematologic Malignancies

Part One Research on an Acute Myeloid Leukemia patientby transcriptome sequencingObjective (1) Screening and identification differentially expressed genes (DEGs) ofacute myeloid leukemia at different stages of the M2type, GO and KEGG pathwayannotation and enrichment analysis were performed on the differentially expressed genesto understand the biological function of differentially expressed genes, and determine itsmain biochemical metabolism and signal transduction pathways participated.(2)Comparing the transcriptome sequences of different stages of AML-M2to the humanreference genome, analyze and identify single nucleotide polymorphism loci and fusiongenes related with disease.Methods The peripheral blood samples of de novo and remission of a patientdiagnosed with acute myeloid leukemia were sequenced by high throughput transcriptomesequencing technology，reads were aligned to the Hg19genome assembly and transcriptsfrom the Ensembl database using TopHat and Bowtie. Transcript abundance anddifferential gene expression were calculated by Cufflinks, GO and KEGG pathwayannotation and enrichment analysis were performed by DAVID. Further analysis theexpression level of AML associated genes such as JAK1, JAK2, TET2, ASXL1, IDH2,WT1, BCR, ABL1, JAK3, GATA2, HOXA9, FLT3, IDH1. Single nucleotide mutations(SNVs) were identified by SAM tools and VarScan by comparison of the AML andremission samples. Through comparing SNVs in de novo and remission to identify thoseassociated with leukemia, further analysis molecular function and metabolic pathways ofthese mutations.Result （1）The pairwise comparison of the AML and remission samples resulted in the identification of a total of4,148DEGs (FDR <0.05),2812of them were downregulated and1336were up-regulated. DEGs were categorized in140GO terms andenriched in26KEGG pathways, and most of them were related to immune response andsignal transduction.WT1, BCR, ABL1, JAK3, GATA2, HOXA9, FLT3, IDH1associatedwith leukemia were DEGs.（2）Through the analysis of SNVs11mutations wererecognized (p <0.05), including ZRSR1, MLXIP, TLN1, LAP3, HK3, et al.Conclusion (1) RNA-Seq provides us with the gene expression patterns of acuteleukemia patients,through the transcriptome comparison analysis between AML-M2diagnosed and complete remission, abnormal expressed genes were discovered inleukemia patients, providing new clues for further explore the pathogenesis of AML.(2)The currence and development of AML has correlation with a variety of geneticabnormalities, may be acquired mutations (MLXIP, HK3, TLN1, ICAM-3), differentiallyexpressed (WT1, BCR, ABL1, JAK3, GATA2, HOXA9, FLT3, IDH1), fusion genes. Inaddition to known genetic abnormalities, more abnormal gene expression has not beenable to be fully aware of. Part Two Significance of BCL6, MYC, P53gene abnormalityon the prognosis of diffuse large B-cell lymphomaObjective To explore how BCL6, MYC, P53gene abnormality influence onDiffuse large B-cell lymphoma(DLBCL) prognostic, and search independent prognosticfactors in DLBCL, provide evidence for clinical prognosis and the selection ofstratification treatment for DLBCL patients.Method65newly diagnosed DLBCL pathological specimens were collected from2009-2012. Interphase fluorescence in situ hybridization technique (I-FISH) was used todetect BCL6, MYC, P53gene status. Combining clinical factors and immunohistochemicalresults multiple-factor survival analysis was performed. Results1.The BCL6rearrangement rate was21.5%(14/65), P53deletion rate was35.4%(23/65), MYC rearrangement rate was7.7%(5/65).2. ALL MYC rearrangement wereGCB subtype, and BCL2protein are all positive, and MYC rearrangement has significantcorrelation with Ki67(P <0.05). BCL6rearrangement has no obvious correlation withBCL6protein expression.3. BCL6rearrangement group has obviously bad overall survival(OS， P=0.027). COX proportional hazards model analysis show that gender, BCL6protein, BCL6rearrangement, Ki67index were prognosis factors independent of IPI.Conclusion1. The BCL6rearrangement has no obvious correlation with BCL6protein.2.BCL6/BCL6influences the prognosis of patients with DLBCL respectively fromthe level of gene and protein, both are independent prognostic factors in DLBCL.3. MYCrearrangement alone has no correlation with the prognosis, it is necessary to evaluate"double hit" impact on prognosis of DLBCL on both gene or protein levels.4. P53deletionhad no obvious effect on the prognosis of patients with DLBCL, combined with literature,mutations in structural domain may be the key factor deciding P53influence on theprognosis of patients with DLBCL.