Adams, David S.
The purpose of this Major Qualifying Project (MQP) was to evaluate a PreservCyt? solution PCR protocol for detecting early stages of lung cancer in human lung specimens. Specific cell lines were obtained from the ATCC, and a collection of cancerous and normal cellular lung specimens were obtained by bronchial washes or induced coughing. These samples were processed into PreservCyt, Cytyc Corporation's proprietary preservative solution that breaks up mucous, red blood cells, and other non-diagnostic debris that may complicate analysis. RNA from the specimens was analyzed by RT-PCR for prohormone convertase 2 (PC2), a known neuroendocrine differentiation marker present in lung carcinomas, and for housekeeper genes. As expected, a PC2 band appeared only in the small cell lung carinoma cell line (positive control sample) and not in non-small cell carcinoma lines or the normal lung fibroblast cell line (negative control sample). Surprisingly, no PC2 bands were detected in the lung specimens obtained from live patients, even those with small cell lung carcinoma. The lack in PC2 signal did not appear to be due to degraded or contaminated specimen RNA as GAPDH, -globin, and Amyloid mRNAs were each successfully amplified from the patient samples. My investigations demonstrate first that the PreservCyt protocol successfully allows isolation of intact RNA from cell lines and patient specimens, and second that PC2 mRNA may be either too rare to amplify or it may not be an adequate marker for the type of lung carcinomas which were tested.
Worcester Polytechnic Institute
Major Qualifying Project
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Biology and Biotechnology