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.