Sensitive detection of small cell lung carcinoma cells by reverse transcriptase–polymerase chain reaction for prepro–gastrin‐releasing peptide mRNA

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Abstract
BACKGROUND Gastrin‐releasing peptide (GRP) is an autocrine growth factor in patients with small cell lung carcinoma (SCLC). The authors developed a reverse transcriptase‐polymerase chain reaction (RT‐PCR) assay for the detection of SCLC cells in the peripheral blood and the pleural effusion using preproGRP mRNA as a target. METHODS The current study was conducted to determine the utility of preproGRP‐specific nested RT‐PCR on the peripheral blood, bone marrow, and pleural effusion samples from 32 patients with SCLC, 39 patients with non‐small cell lung carcinoma (NSCLC), 28 patients with nonmalignant pulmonary disease, and 20 healthy volunteers. The internal primers were designed to amplify a 244‐base pair PCR product, a sequence encompassing exon 1 and exon 2 by the nested RT‐PCR assay. RESULTS Amplification of the preproGRP message was detected in SCLC cell lines (LU165, SBC1, SBC2, and SBC3) but not in other NSCLC cell lines (A549, ABC1, EBC1, and Oka‐1). The SCLC cells (LU165) were detected in dilutions of tumor cells of up to 10−7 in hematopoietic cells from healthy donors. The preproGRP mRNA was detected in 16 of 32 (50%) blood samples, 2 of 11 (18%) marrow samples, and in all 6 (100%) pleural effusion samples. Blood samples gave positive results in 11 of 19 (58%) patients with extensive disease compared with 5 of 13 (38%) patients with limited disease. In contrast, only 1 blood sample (2.6%) from a patient with lung adenocarcinoma gave a positive result among patients with NSCLC. No other samples of blood, bone marrow, and pleural effusion from patients with NSCLC and none of the blood samples from patients with nonmalignant diseases and healthy volunteers were positive. CONCLUSIONS The current RT‐PCR approach may be a sensitive and specific assay to detect SCLC cells in circulating blood as well as in pleural effusions from SCLC patients. Cancer 2003;97:2504–11. © 2003 American Cancer Society. DOI 10.1002/cncr.11378

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