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Quantitative detection of circulating epithelial cells by Q-RT-PCR
Vladimir V Iakovlev1, Rashmi S Goswami, Jonathan Vecchiarelli
1CIHR Molecular Pathology of Cancer Fellowship Program, Ontario Cancer Institute/Princess Margaret Hospital, Toronto, Ontario, Canada.
Breast Cancer Research and Treatment
|March 10, 2007
Summary
Quantifying circulating tumor cells (CTCs) using real-time quantitative reverse transcriptase PCR (Q-RT-PCR) is feasible. This method reliably detects as few as 5 cells per sample, aiding breast cancer patient monitoring.
Area of Science:
- Oncology
- Molecular Biology
- Biotechnology
Background:
- Circulating tumor cells (CTCs) are crucial biomarkers for predicting survival and treatment outcomes in breast cancer patients.
- Real-time quantitative reverse transcriptase PCR (Q-RT-PCR) offers high sensitivity for detecting CTCs.
Purpose of the Study:
- To evaluate the utility of Q-RT-PCR for quantifying CTCs in breast cancer.
- To assess the expression and amplification linearity of specific genes for CTC detection.
Main Methods:
- Screened multiple genes (CK19, maspin, mammaglobin, GAPDH, RPL19) for expression and amplification linearity using breast cancer cell lines.
- Simulated CTCs by spiking cultured cells into healthy volunteer blood, followed by enrichment, RNA extraction, and Q-RT-PCR.
Main Results:
- Cytokeratin 19 (CK19) demonstrated reliable expression and linear amplification across tested cell lines.
- The Q-RT-PCR method achieved a lower limit of reliable detection at 5 cells per sample, equivalent to 0.7 cells/ml in 7.5 ml of blood.
Conclusions:
- Q-RT-PCR is a viable technique for quantifying CTCs in breast cancer.
- This method supports high-throughput comparative analysis for patient monitoring during treatment and follow-up.
