Identification of genes showing differential expression in antisense K-ras-transduced pancreatic cancer cells with suppressed tumorigenicity

K-ras point mutation occurs in >80% of pancreatic cancer. We reported previ ously that the transduction of an antisense K-ras RNA expression vector sup pressed the growth of pancreatic cancer cells with K-ras point mutations in vitro and in vivo. The RNA differential display method (DD) was used to co mpare the mRNA expression profile of the pancreatic cancer cell line AsPC-1 and that of the antisense K-ras-transduced, growth-retarded AsPC-1 cells. cDNA fragments were isolated from 20 bands on the DD gel, and their differe ntial expression between the two cell lines was confirmed, A sequence analy sis revealed that all of the 11 clones up-regulated in the antisense-transd uced cells were mitochondrial genes. The other nine cDNA clones that were d own-regulated in the antisense-transduced AsPC-1 cells included an oncogene PTI-1 (prostate tumor inducing gene-1), matrix metalloproteinase (MMP)-7, the beta 3 chain of laminin-5, lysosome-associated membrane protein-2, the Il chain of apoferritin, ribosomal protein S6, proteasome submit XAPC7, and two cDNA fragments with no homology to the GenBank database. In addition t o the AsPC-1 cells, reverse transcription-PCR analysis on surgical specimen s of pancreatic cancer revealed that the PTI-1 and MMP-7 genes were overexp ressed in three and four cases, respectively, of five cases examined. This method offers a unique opportunity to identify a set of genes that may be m odulated by K-ras activation, at Least in a subset of the pancreatic cancer . The information on such genes may facilitate our understanding of the spe ctrum of the functional genetic changes in pancreatic cancer.