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Using Suppression Subtractive Hybridization Technique to Clone Differentially Expressed Genes between Renal Cell Carcinoma and the Matched Normal Renal Epithelium

Author: DongMeiZuo
Tutor: ChangJiWu
School: Tianjin Medical University
Course: Department of Urology
Keywords: Renal cell carcinoma Suppression subtractive hybridization Differential gene CDNA library
CLC: R737.11
Type: Master's thesis
Year: 2004
Downloads: 77
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Abstract


Highly malignant renal cell carcinoma, clinical manifestation, the incidence of systemic cancer accounted for about 3%. Worldwide incidence of renal cell carcinoma increased by 2% per year, renal cell carcinoma who die each year from nearly 100,000 cases. In China, the incidence of renal cell carcinoma of urinary tract tumors accounted for the second place, there is a rising trend in recent years, a serious threat to people's health. Involved in the regulation of cell growth, differentiation and death of a large number of genes, to ascertain its exact mechanism of occurrence and development of renal cell carcinoma-specific needs expressed genes were cloned and analyzed to understand their gene expression profiles. This is the future of renal cell carcinoma-specific diagnosis, treatment and prognosis have biological significance. Cloning of differentially expressed genes in many ways, Diatchenko established suppression subtractive hybridization technique is in many ways a more prominent advantages. Its high specificity, high sensitivity, low false positive, the process is relatively simple. In this study, using suppression subtractive hybridization (Suppression Subtractive Hybridization, SSH) technology to build a human renal cell carcinoma and corresponding normal kidney epithelial tissue differentially expressed cDNA subtractive library, and use dot blotting (Dot Blot) screened out renal cell technology Cancer differentially expressed genes, some randomly selected positive clones were sequenced, the results do in GenBank homology analysis. Tianjin Medical Journal study results show that: The application SSH technology to build high abatement efficiency of human renal cell carcinoma and corresponding normal renal epithelial tissue subtractive cDNA library, library contains 400 positive clones were randomly picked 200 clones were amplified, Dot blot screening 177 positive clones, 55 clones of the CDNA insert fragments were sequenced after the GenBank database shows contains 26 kinds of independent gene fragment gene 24 kinds of known and unknown two kinds of gene fragments. This study was to construct human renal cell carcinoma and normal renal epithelium difference subtractive cDNA libraries, both received some fragments of known cancer-related genes, validated results of previous studies; also won a number of new genes for further study gene fragments function basis. But also for the future of renal cell carcinoma-specific diagnosis, prognosis and biological therapy provides a new theoretical basis. Human renal cell carcinoma subtractive library was constructed successfully showed suppression subtractive hybridization technique is screening and cloning of differentially expressed genes in renal cell carcinoma, an effective means of making large quantities of further screening and cloning of genes specifically expressed in renal cell carcinoma is possible to elucidate renal cancer cells, the molecular mechanism of the development of new research methods.

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CLC: > Medicine, health > Oncology > Genitourinary tumors > Urinary tumors > Kidney,renal pelvis tumor
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