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Epidermal growth factor receptor mRNA in peripheral blood of patients with pancreatic, lung, and colon carcinomas detected by RT-PCR.
Int J Oncol. 2003 Feb; 22(2):425-30.IJ

Abstract

The epidermal growth factor receptor (EGFR) is overexpressed in many solid tumors. An RT-PCR assay was used to detect EGFR mRNA in the blood of 43 patients with pancreatic, lung (NSCLC), colon, and renal carcinomas. After an initial serum tube was drawn to clear the line of detached epithelial cells, a 3 ml sample of EDTA-anticoagulated blood was collected from each cancer patient and healthy control. Total RNA was isolated from each sample and from cancer cell lines. After reverse transcription using specific priming, outer and nested primers for EGFR were employed for cDNA amplification. RNA integrity was confirmed with RT-PCR amplification using beta2-microglobulin primers. PCR products were electrophoresed on agarose gels containing ethidium bromide and visualized. The assay was validated using Southern blotting and was capable of detecting a lower limit of 100 fg of total RNA from the A431 cell line. EGFR-positive cells were detected in 3/10 (30%) non-small cell lung cancer patients, 2/11 (18%) pancreatic cancer patients, and 2/16 (12.5%) colon cancer patients, but in 0/6 (0%) patients with renal carcinoma (16% of solid tumor patients overall). All 23 healthy controls were negative. This study is the first to apply RT-PCR for the detection of EGFR mRNA in the peripheral blood of patients with pancreatic and renal carcinomas, and it lends further support for the use of EGFR mRNA as a marker of CTCs in the blood of patients with certain types of solid tumors.

Authors+Show Affiliations

The Pennsylvania State University College of Medicine/Hershey Medical Center, Hershey, PA 17033, USA. lclarke816@aol.comNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Evaluation Study
Journal Article

Language

eng

PubMed ID

12527944

Citation

Clarke, Loren E., et al. "Epidermal Growth Factor Receptor mRNA in Peripheral Blood of Patients With Pancreatic, Lung, and Colon Carcinomas Detected By RT-PCR." International Journal of Oncology, vol. 22, no. 2, 2003, pp. 425-30.
Clarke LE, Leitzel K, Smith J, et al. Epidermal growth factor receptor mRNA in peripheral blood of patients with pancreatic, lung, and colon carcinomas detected by RT-PCR. Int J Oncol. 2003;22(2):425-30.
Clarke, L. E., Leitzel, K., Smith, J., Ali, S. M., & Lipton, A. (2003). Epidermal growth factor receptor mRNA in peripheral blood of patients with pancreatic, lung, and colon carcinomas detected by RT-PCR. International Journal of Oncology, 22(2), 425-30.
Clarke LE, et al. Epidermal Growth Factor Receptor mRNA in Peripheral Blood of Patients With Pancreatic, Lung, and Colon Carcinomas Detected By RT-PCR. Int J Oncol. 2003;22(2):425-30. PubMed PMID: 12527944.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Epidermal growth factor receptor mRNA in peripheral blood of patients with pancreatic, lung, and colon carcinomas detected by RT-PCR. AU - Clarke,Loren E, AU - Leitzel,Kim, AU - Smith,Jill, AU - Ali,Suhail M, AU - Lipton,Allan, PY - 2003/1/16/pubmed PY - 2003/8/27/medline PY - 2003/1/16/entrez SP - 425 EP - 30 JF - International journal of oncology JO - Int J Oncol VL - 22 IS - 2 N2 - The epidermal growth factor receptor (EGFR) is overexpressed in many solid tumors. An RT-PCR assay was used to detect EGFR mRNA in the blood of 43 patients with pancreatic, lung (NSCLC), colon, and renal carcinomas. After an initial serum tube was drawn to clear the line of detached epithelial cells, a 3 ml sample of EDTA-anticoagulated blood was collected from each cancer patient and healthy control. Total RNA was isolated from each sample and from cancer cell lines. After reverse transcription using specific priming, outer and nested primers for EGFR were employed for cDNA amplification. RNA integrity was confirmed with RT-PCR amplification using beta2-microglobulin primers. PCR products were electrophoresed on agarose gels containing ethidium bromide and visualized. The assay was validated using Southern blotting and was capable of detecting a lower limit of 100 fg of total RNA from the A431 cell line. EGFR-positive cells were detected in 3/10 (30%) non-small cell lung cancer patients, 2/11 (18%) pancreatic cancer patients, and 2/16 (12.5%) colon cancer patients, but in 0/6 (0%) patients with renal carcinoma (16% of solid tumor patients overall). All 23 healthy controls were negative. This study is the first to apply RT-PCR for the detection of EGFR mRNA in the peripheral blood of patients with pancreatic and renal carcinomas, and it lends further support for the use of EGFR mRNA as a marker of CTCs in the blood of patients with certain types of solid tumors. SN - 1019-6439 UR - https://www.unboundmedicine.com/medline/citation/12527944/Epidermal_growth_factor_receptor_mRNA_in_peripheral_blood_of_patients_with_pancreatic_lung_and_colon_carcinomas_detected_by_RT_PCR_ L2 - http://www.spandidos-publications.com/ijo/22/2/425 DB - PRIME DP - Unbound Medicine ER -