Detection of colon carcinoma cells in blood of
patients using RT-PCR and GCC-specific nested primers. Total RNA was
extracted from a 1-ml aliquot of blood cells, mRNA-purified, and used
for RT-PCR, using GCC-specific nested primers, as described. Border
lanes (M) indicate size markers (200–1000 bp). Arrow indicates the
predicted size of the amplification product for human GCC (hGCC; ≈250
bp). Lane 1, normal volunteers (n = 10); lane 2,
patients with a colonic adenomatous polyp (n = 6);
lane 3, patient with an acute exacerbation of inflammatory bowel
disease (n = 1); lane 4, patient with
adenocarcinoma of the lung (n = 1); lane 5, patient
with adenocarcinoma of the breast (n = 1); lane 6,
patient with adenocarcinoma of the ovary (n = 1);
lane 7, patient with lymphoma (n = 1); lane 8,
patients with pancreatic carcinoma (n = 6); lanes
9–11, patients with Dukes A colorectal cancer; lanes 12–16, patients
with Dukes B colorectal cancer; lanes 17–20, patients with Dukes C
colorectal cancer; and lanes 21–30, patients with Dukes D colorectal
cancer.