Abstract: Objective：To elucidate the smad4-independent signaling pathway of TGF-β 1in the pancreatic cell carcinogenesis Methods: We transfected pancreatic cancer cell line, BxPC3 which carries homozygosity loss of SMAD4 , with TGF-β1 and pcDNA3 as a control. Cell growth was detected by flow cytometry (FCM) and expression of TGF-β1 detected by western blot. By the technique of suppression subtractive hybridization (SSH), we constructed a substracted cDNA library of TGF-β1 related genes. Amplification of the library was carried out with the E. coli strain JM109. Reverse northern blot was used to confirm the genes differentially expressed after TGF-β 1 functioned. Results: The growth of BxPC3 was slightly inhibited after transfected with TGF-β and overexpression of this cytokine was convinced by western blot. TGF-β 1 related subtractive library with high subtractive efficiency was set up successfully. The amplified library contained 300 positive clones. Reverse northern blot showed that 32 clones were actually differentially expressed. After sequencing and blastn searching, we found 10 genes up-regulated and 12 down-regulated,13 of which habour known function and 9 unknown. Conclusions: With this subtracted cDNA library, it is easy for us to reveal the molecular mechanism in the pancreatic carcinogenesis.