Detection of gastrin mRNA in fresh human colonic carcinomas by reverse transcription-polymerase chain reaction.
To investigate the hypothesis that gastrin might be synthesized by tumour tissues in cancer of the colon, samples from six human colon tumours, one hepatic metastasis, four normal colonic mucosal samples and two antral and one fundic gastric mucosal samples from nine patients were analysed to determine whether gastrin mRNA was present. RNA was extracted from surgical specimens by ultracentrifugation on a CsCl cushion, purified using the guanidinium thiocyanate method, reverse-transcribed and amplified by polymerase chain reaction. Gastrin mRNA was detected in each colonic carcinoma sample (including the hepatic metastasis), while no such signal was observed in normal colon biopsies. Positive and negative controls (gastric antrum and fundus respectively) gave the expected results. In each of the positive samples, the chemiluminescent revelation of amplified products after Southern blotting corresponded to gastrin mRNA without the intron. These findings demonstrate the ability of primary and metastatic human colonic tumours to produce gastrin mRNA. Since malignant cell lines have been reported to produce gastrin peptide, and since gastrin receptors were present in some cases, our results support the idea that gastrin may be involved in an autocrine mechanism.
Monges
G
G
Paoli-Calmettes Institute, Marseille, France.
Biagini
P
P
Cantaloube
J F
JF
Chicheportiche
C
C
Frances
V
V
Brandini
D
D
Parc
P
P
Seitz
J F
JF
Giovannini
M
M
Sauvan
R
R
D001483:Base Sequence; D003106:Colon / Q000737:chemistry; D003110:Colonic Neoplasms / Q000737:chemistry / Q000235:genetics*; D005748:Gastric Fundus / Q000737:chemistry; D005753:Gastric Mucosa / Q000737:chemistry; D005755:Gastrins / Q000235:genetics*; D006801:Humans; D007413:Intestinal Mucosa / Q000737:chemistry; D008113:Liver Neoplasms / Q000737:chemistry / Q000235:genetics / Q000556:secondary; D008163:Luminescent Measurements; D008969:Molecular Sequence Data; D009363:Neoplasm Proteins / Q000235:genetics*; D009928:Organ Specificity; D016133:Polymerase Chain Reaction; D011706:Pyloric Antrum / Q000737:chemistry; D012333:RNA, Messenger / Q000032:analysis* / Q000235:genetics; D012194:RNA-Directed DNA Polymerase
D005755:Gastrins; D009363:Neoplasm Proteins; D012333:RNA, Messenger; D012194:RNA-Directed DNA Polymerase