Molecular detection of metastatic pancreatic carcinoma cells using a multimarker reverse transcriptase-polymerase chain reaction assay

BACKGROUND, The diagnosis of pancreatic carcinoma is often associated with a poor prognosis, because most patients already have advanced disease. A hi ghly sensitive assay to detect the progression of pancreatic carcinoma woul d be of significant clinical utility. The authors developed multiple tumor mRNA markers for reverse transcriptase-polymerase chain reaction (RT-PCR) t o detect metastatic tumor cells in the blood and tissue of patients with Am erican Joint Committee on Cancer (AJCC) Stage II/III or IV pancreatic carci noma. METHODS. An RT-PCR plus Southern blot assay was used to detect mRNA of tumo r markers in blood and tissues. mRNA expression of the tumor progression ma rkers MET (hepatocyte growth factor receptor gene c-met), GalNAc-T (beta 1, 4- N-acetylgalactosaminyl-transferase), and beta-hCG (beta-human chorionic gonadotropin) was evaluated in 9 pancreatic carcinoma cell lines, 13 tumor biopsy specimens, 5 nonmalignant pancreatic tissue specimens, and blood fro m 33 pancreatic carcinoma patients and 32 healthy donors. RESULTS, The detection limit of the assay was 1 rho g, 10 rho g, and 10 rho g for MET, GalNAc-T, and beta-hCG mRNA expression, respectively. The pancr eatic carcinoma cell lines expressed all three mRNA markers. Of blood speci mens from 17 patients with AJCC Stage TV pancreatic carcinoma, 82%, 65%, an d 76% were MET, GalNAc-T, and P-hCG mRNA positive, respectively. Of blood s pecimens from 16 patients with ATCC Stage II/III disease, 88% were positive for at least 1 mRNA marker. CONCLUSIONS. A multiple molecular marker assay was developed to detect canc er cells in blood and tissue from patients with different stages of pancrea tic carcinoma. The detection of cancer cells in the blood may be used as a marker of pancreatic tumor progression and may be useful in monitoring resp onse to therapy. (C) 2000 American Cancer Society.