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. 2011 Oct 4:11:422.
doi: 10.1186/1471-2407-11-422.

Gene expression profile of circulating tumor cells in breast cancer by RT-qPCR

Areti Strati  1 , Athina Markou, Cleo Parisi, Eleni Politaki, Dimitris Mavroudis, Vasilis Georgoulias, Evi Lianidou
Affiliations

Gene expression profile of circulating tumor cells in breast cancer by RT-qPCR

Areti Strati et al. BMC Cancer. .

Abstract

Background: Circulating tumor cells (CTCs) have been associated with prognosis especially in breast cancer and have been proposed as a liquid biopsy for repeated follow up examinations. Molecular characterization of CTCs is difficult to address since they are very rare and the amount of available sample is very limited.

Methods: We quantified by RT-qPCR CK-19, MAGE-A3, HER-2, TWIST1, hTERT α+β+, and mammaglobin gene transcripts in immunomagnetically positively selected CTCs from 92 breast cancer patients, and 28 healthy individuals. We also compared our results with the CellSearch system in 33 of these patients with early breast cancer.

Results: RT-qPCR is highly sensitive and specific and can detect the expression of each individual gene at the one cell level. None of the genes tested was detected in the group of healthy donors. In 66 operable breast cancer patients, CK-19 was detected in 42.4%, HER-2 in 13.6%, MAGE-A3 in 21.2%, hMAM in 13.6%, TWIST-1 in 42.4%, and hTERT α+β+ in 10.2%. In 26 patients with verified metastasis, CK-19 was detected in 53.8%, HER-2 in 19.2%, MAGE-A3 in 15.4%, hMAM in 30.8%, TWIST-1 in 38.5% and hTERT α+β+in 19.2%. Our preliminary data on the comparison between RT-qPCR and CellSearch in 33 early breast cancer patients showed that RT-qPCR gives more positive results in respect to CellSearch.

Conclusions: Molecular characterization of CTCs has revealed a remarkable heterogeneity of gene expression between breast cancer patients. In a small percentage of patients, CTCs were positive for all six genes tested, while in some patients only one of these genes was expressed. The clinical significance of these findings in early breast cancer remains to be elucidated when the clinical outcome for these patients is known.

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Figures

Figure 1
Figure 1
Schematic diagram of the entire analytical procedure.
Figure 2
Figure 2
Quantification of CK-19, MAGE-A3, HER-2, TWIST-1, hMAM (Mammaglobin), and h-TERT α+β+ transcripts in CTCs (copies/mL of peripheral blood).
Figure 3
Figure 3
Heat map of CK-19, MAGE-A3, HER-2, TWIST-1, hMAM (Mammaglobin), and hTERT α+β+ expression in CTCs as quantified by RT-qPCR.
Figure 4
Figure 4
Comparison of RT-qPCR and CellSearch in early breast cancer patients (n = 33).
See this image and copyright information in PMC

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