Identification of genes with specific expression in pancreatic cancer by cDNA representational difference analysis

Department of Internal Medicine I, University of Ulm, Germany.
Genes Chromosomes and Cancer (Impact Factor: 4.04). 07/1997; 19(2):97-103. DOI: 10.1002/(SICI)1098-2264(199706)19:23.0.CO;2-V
Source: PubMed


cDNA representational difference analysis (cDNA-RDA) is a polymerase-chain-reaction-coupled subtractive and kinetic enrichment procedure for the isolation of differentially expressed genes. In this study, the technique was used to isolate novel genes specifically expressed in pancreatic cancer. cDNA-RDA was done on cDNA reverse transcribed from a poly(A)+ mRNA pool made from 10 cancer tissues (tester) by using as a driver a cDNA from a poly(A)+ mRNA pool made from a combination of 10 tissues of chronic pancreatitis and 10 healthy pancreatic tissues. The use of chronic pancreatitis in addition to healthy pancreas mRNA in the driver preparation eliminated the influence of stromal tissue components present as contamination in the cancer-specific preparations. Such cDNA-RDA led to the isolation of 16 distinct, cancer-specific gene fragments. These were confirmed to be overexpressed in pancreatic cancer tissues by Northern blot analysis. Sequence analysis revealed homologies to five genes previously implicated in the carcinogenesis of the pancreas or other tissues. Eleven fragments had no significant homology to any known gene and thus represent novel candidate disease genes. The experiments demonstrate that cDNA-RDA is a reproducible and highly efficient method for the identification of novel genes with cancer-specific expression.

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    • "TM4SF5 (20,823 Da) is a transmembrane glycoprotein; as a family group, it is related to the tetraspanin family (transmembrane 4 L six family), including TM4SF1 (L6, L6-Ag), TM4SF4 (IL-TIMP), TM4SF518 (L6D), and TM4SF20 [61, 62]. TM4SF5 is highly expressed in diverse types of cancers, including liver, pancreatic, gastric, colon, adrenocorticotropic hormone (corticotropin)-negative bronchial carcinoid tumors, soft-tissue sarcoma, nonendocrine lung, and papilla vateri carcinoma [63, 64, 65, 66]. Similar to tetraspanins (i.e., transmembrane 4 superfamily, TM4SFs), TM4SF5 has four transmembrane domains (TM1-TM4), short cytoplasmic domains at their N- and C-termini, an ICL between TM2 and TM4, two extracellular loops, a smaller extracellular loop between TM1 and TM2, and a larger extracellular loop between TM3 and TM4 [61, 62]. "
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    03/2014; 12(1):12-20. DOI:10.5808/GI.2014.12.1.12
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    • "Comparing expression profiles from primary tumors with samples from histologically non-malignant pancreatic tissue and chronic pancreatitis (CP) we identified a 147-gene signature correlated with primary pancreatic tumor. This strategy was devised to favor the identification of tumor specific markers rather than transcripts associated with the stromal cell component, which is augmented in both tumor and CP samples [36,37]. We sought to validate the pancreatic cancer expression signature by performing a meta-analysis with published gene expression studies of pancreatic cancer. "
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    Molecular Cancer 11/2011; 10(1):141. DOI:10.1186/1476-4598-10-141 · 4.26 Impact Factor
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    • "Microarray analyses have consistently found it to be one of the most upregulated genes in advanced prostate cancer (Dhanasekaran et al., 2001; Magee et al., 2001). Other transmembrane serine proteases implicated in cancer progression are membrane-type serine protease-1/matriptase in breast cancer (Shi et al., 1993), TMPRSS2 in prostate cancer (Afar et al., 2001) and TMPRSS4 in pancreatic cancer (Gress et al., 1997). "
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