Preparation Of Lymphoma-specific Oligonucleotide Microarray And Its Application In Exploring The Gene Expression Profiles Of Lymphoma Lineage Cells

The morbific mechanism of tumor is very complicated, and the tumor-related genes play an important role in the occurrence, development, diagnosis, therapy, and prognosis of tumors. It is important to understand the tumor-related genes" construction, function, expression, and the molecular mechanism of cell cycle control and apoptosis regulation. DNA chip technology can rapidly, massively and parallelly monitor and analyze a large number of gene expressions. It will play an important role in exploring the molecular mechanism of tumors to establish DNA microarray technology platform.Purpose: Application the oligonucleotide microarray technology to detect the gene expression profiles of the lineage cells of BL and CTCL, identify the differentially expressed genes between 2 different kinds of cells, investigate the molecular mechanism of lymphoma, and promote the clinical diagnosis and therapy for it.Steps and Methods:1)Preparation of lymphoma-related genes microarray: 116 lymphoma-related genes were involved in the study. By using of a self-made software system, oligonucleotide probes were designed and Blast. All the probes have a very high specificity, i.e. except target sequence, the similarity between the probe and non-target sequences is less than 70%. In the probes with the same length 40, GC content are in a narrow scope (from 45% to 55%). All the probes are modified with amino at 5'or 3'terminal. The optimal concentration is 0.5i g/i 1. The oligonucleotide probes were synthesized and spotted onthe chemical-material-coated-glass plates in array.2)Application of lymphoma-related genes microarray in studying lymphoma - specific genes from malignant lymphoma-cell/normal lymphocytes: The total RNAs were isolated from Burkitf s lymphoma lineage cells, cutaneous T lymphoma lineage cells, and normal lymphocytes, and they were reversely transcribed to cDNAs with the incorporations of fluorescent dUTP-Cy3 or dUTP-Cy5 to prepare the hybridization probes. The mixed probes were hybridized to the oligonucleotide microarray. After high-stringent washing, the microarray was scanned for the fluorescent signals and showed differences between 2 cells, and preliminarily analyzed the function of the differentially expressed genes by bioinformatics.Results: In Burkitf s lymphoma lineage cells, we identified 7 up-regulated and 4 down-regulated genes. In cutaneous T lymphoma lineage cells, we identified 6 up-regulated and 2 down-regulated genes. And preliminaryly analyzed the function of the differentially expressed genes by bioinformatics, we find that these genes are intensively related to cell cycle control, signal transduction, apoptosis regulation and differentiation.Table 1. the differentially expressed genes between lymphoma-cell/normal lymphocytesConclusion: The oligonucleotide microarray technology is an effective technique in screening for differentially expressed genes between 2 different kinds of cells. The expression of many genes changed in the malignant lymphoma. Further analysis of the obtained genes will help to understand the molecular mechanism of lymphoma, and also may promote the diagnosis, identification of new targets for specific treatmentapproaches for it.