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《Acta Academiae Medicinae Militaris Tertiae》 2007-04
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Screening and identification of a novel gene-related human pancreatic cancer

JIANG Ye-gui 1,LI Zhao-shen 2,WANG Yu-ming 1,LIU Tong-hua 3(1Institute of Infectious Diseases,Southwest Hospital,Chongqing 400038,3Department of Pharmacy,Xinqiao Hospital,Third Military Medical University,Chongqing 400037,2Department of Gastroenterology,Changhai Hospital,Second Military Medical University,Shanghai 200433,China)  
Objective To screen and identify a novel gene-related human pancreatic cancer.Methods Total RNA and mRNA were isolated from human pancreatic cancer(as tester)and normal paracancerous pancreas tissues(as driver),respectively.The suppressive subtractive cDNA library was set up using suppressive subtractive hybridization(SSH).The positive clones randomly selected in the library were sequenced and analyzed for homology with BLAST.The expression of the novel gene in 12 specimens of pancreatic cancer and normal tumor-surrounding pancreas tissues and 4 pancreatic cancer cell lines(PaTu8988,SW1990,BxPC-3,AsPC-1)were detected by RT-PCR and Northern blot analysis.Results Random analysis of 50 white clones with PCR amplification showed that 47 clones contained inserted fragments.The positive rate was 94%.The 47 positive clones were selected for further sequencing and 42 usable sequences were obtained.After BLAST homology analysis,41 of them were found to be known genes,and one of them was found to be unknown gene with no functional clues,might being a novel gene.RT-PCR analysis showed that the novel gene expressed in 12 specimens of pancreatic cancer tissues and 4 pancreatic cancer cell lines,but not in normal pancreas tissues.This result was just the same as the result of Northern blot analysis.Conclusion A suppressive subtractive cDNA library of human pancreatic cancer is constructed successfully by SSH.One novel gene is found and might be novel pancreatic cancer-associatedgene.The novel gene is associated with oncogenesis and development of pancreatic cancer,and might be potential tumor markers or molecular target for diagnosis and treatment of pancreatic cancer.
【CateGory Index】: R735.9
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