关键词: Notch信号通路, 胰腺癌, RNA干扰, Jagged2基因

Abstract: Objective The Objective of this study was to investigate the abnormal expression of Notch signaling pathway members in pancreatic cancer and its important effect on the development of pancreatic carcinoma.Methods Affymetrix gene expression microarray was used to screen the differentially expressed members of Notch signal pathway in 10 cases of pancreatic carcinoma and its adjacent tissues,and verified by real-time quantitative PCR and Western blotting.The lentivirus expression vector carrying the siRNA fragment of Jagged 2(JAG2)gene was transfected into the pancreatic cancer primary cells to construct the JAG2 gene repression-expressing pancreatic cancer cell line.MTT,flow cytometry and Transwell assays were used to analyze cell proliferation,changes of cell cycle and invasive transfer capabilities.Results A total of 512 differentially expressed genes were detected by Affymetrix gene expression microarray,including 419 up-regulated genes and 93 down-regulated genes.JAG2(up-regulated expression 8.20 times),NOTCH1(up-regulated expression 3.74 times),HES1(up-regulated expression 3.27 times),and NOTCH2(up-regulated expression 3.16 times)were differentially expressed in Notch signaling pathway.The results of PCR and Western blotting were consistent with those of gene chip.The growth curves of JAG2 gene repressed pancreatic cancer cells and pancreatic cancer primary cells were drawn by the standard OD490 value of d1-d5 by MTT assay.JAG2 gene repressor expression vector could significantly inhibit the proliferation of pancreatic cancer cells.The cell cycle analysis showed that the apoptosis and the arrested cell cycle at the S phase were significantly increased in pancreatic cancer cells with JAG2 gene repressor expression.The invasive ability was significantly reduced in JAG2 gene repressor expression pancreatic cancer cells(P＜0.05).Conclusion Some members of the Notch signaling pathway are significantly differentially expressed in pancreatic cancer tissues,and repression of this member can affect the growth,cell cycle,invasion and metastasis of pancreatic cancer cells.

Key words: Notch signaling pathway, Pancreatic cancer, RNA interference, Jagged 2 gene