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[Application of subtractive hybridization in screening for colorectal cancer negatively related genes]
1Cancer Institute and Medical Molecular Biology Center, Zhejiang Medical University, Hangzhou.
Objective:
To search previously uncertained factors that corelated with the occurence and development of human colorectal cancer.
Method:
Subtractive hybridization technique was employed to screen down-regulated genes of colorectal cancer.
Results:
We obtained 46 clones which highly were expressed in normal intestinal mucosa but much lower or none in colorectal cancer. Two of the 46 clones, SCN6 and SCN19, were accepted by the Genbank Database as novel genes with the accession numbers U17714 and U20248. Full length cDNA (2932bp) of SNC6 was sequenced, and supposed to encode a protein comprised 271 amino acids, MW30kd, which shared less than 30% homology with the protein database. The gene was localized to human chromosome 22q13. RNA dot blot analysis showed that the expression level of SNC6 was lower in colorectal cancer than in normal intestinal mucosa, and similar results were found in breast cancer. Northern blot showed higher expression levels of SNC6 in liver, lung, prostate, testis, ovary and K562 cell line than in other tissues.
Conclusion:
SNC6 can be further investigated as a novel colorectal cancer negatively related gene.