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Use of Standardized Reverse Transcription-Polymerase Chain Reaction and the Standardized Expression Measurement Center in Multi-institutional Trials To Develop Meaningful Lung Cancer Classification Based on Molecular Genetic Criteria^*

^* From the Department of Medicine, Medical College of Ohio, Toledo, OH.

Correspondence to: James C. Willey, MD, George Isaac Professor for Cancer Research, Rm 0012, Ruppert Health Bldg, Medical College of Ohio, 3000 Arlington Ave, Toledo, OH 43614; e-mail: jwilley{at}mco.edu

Through the Director’s Challenge Program of the National Cancer Institute, we have joined with others in an effort to establish a more meaningful classification of lung cancer based on molecular genetic criteria. Specifically, this program has funded our laboratory in developing and implementing the standardized reverse transcription (StaRT) polymerase chain reaction (PCR) method. StaRT-PCR allows numerical, standardized measurement of hundreds of genes to be made simultaneously. Using this method, data from multiple experiments and laboratories may be entered into the same databank, thereby facilitating multi-institutional trials. We have developed software that allows the rapid sorting of data by samples according to annotated criteria and the averaging of the gene expression values among the sorted group of samples. This will enable the establishment of gene expression values for important classes, and this, in turn, will allow the identification of diagnostic criteria.

We have optimized the methods for the measurement of gene expression by StaRT-PCR in cytologic specimens obtained by transthoracic fine-needle aspiration or bronchoscopy. As an example, using these methods we have identified an index comprising the product of c-myc and E2F1 gene expression values divided by the p21 expression value that is 100% accurate for diagnosing lung cancer in cytologic specimens. This is an improvement over the 80% sensitivity rate for the diagnosis of lung cancer in cytologic samples by morphologic criteria.

Through National Cancer Institute Shared Resources Program funding, we have established a Standardized Expression Measurement (SEM) Center. In the SEM Center, StaRT-PCR gene expression measurement is automated and data are automatically collected into the standardized expression database. Currently, we are measuring the expression of c-myc, E2F1, and p21 genes along with > 30 genes associated with poor survival, and 6 genes putatively associated with chemoresistance in a series of lung adenocarcinoma samples that have been compared to matched normal samples. The goal of these studies is to establish a clinical test comprising < 30 genes that will accurately diagnose lung cancer and will predict the optimum treatment for each individual tumor at the time of the initial biopsy.

	Footnotes

 
Abbreviations: PCR = polymerase chain reaction; SEM = Standardized Expression Measurement; StaRT = standardized reverse transcription

Dr. Willey and Ms. Crawford have a significant interest in Gene Express, Inc, which produces and markets StaRT-PCR reagents.

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