Expression of Gut Hormone Gene in Gastric Mucosa Obtained by Endoscopic Biopsy

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We developed a plain polymerase chain reaction (PCR) technique sensitive enough to detect gastrin mRNA in gastric antral mucosa obtained from the patients of peptic ulcer by endoscopic biopsy. From only 1 mug of total cellular RNA we can detect gastrin mRNA.Gastric antral mucosas were obtained by endoscopic biopsy in the patients with peptic ulcers. We can obtain 20 mug of total cellular RNA form 40 mg of gastric antral mucosa. We chose beta-actin as our internal control because, as a major component of the cytoskeletal system, it is transcribed in most eukaryotic cells. Northern blot hybridizations for gastrin mRNA and beta-actin were carried out using synthetic oligodeoxyribonucleotides. Beta-actin mRNAs (2.0 kb) were detected in all specimens examined, but gastrin mRNAs (0.5 kb) was detected in half of them by Northern hybridization. We used 1 mug of total RNA in our gastrin PCR assay. Total cellular RNA was treated with DNase to remove any contaminating DNA. We also chose beta-actin as internal control of amplification. Gastrin and beta-actin transcripts can be simultaneously amplified from gastric antral mucosa. The ratio of PCR-amplified gastrin to beta-actin signal was well correlated with the ratio of gastrin to beta-actin signal in Northern blot hybridization.