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Patient blood samples positive for CTCs were analysed for EMT markers (Twist1, Akt and PI3Kα) and the BCSC marker aldehyde dehydrogenase 1, a detoxifying enzyme responsible for the oxidation of intracellular aldehydes [ 77].
In our samples, ALDH1 overexpression was found in approximately 70% of the blood samples also positive for CTCs.
Using such a threshold, 14 of 27 patient samples were positive for CTCs (52%).
There was no significant correlation (P = 0.065) in this case as well, since there were 53 samples found positive for CTC by the multiplex RT-qPCR assay but negative by the AdnaTest and 37 samples found positive for CTC by the AdnaTest but negative by the multiplex RT-qPCR assay (Table 2).
Here we demonstrated that at least one or more of the EMT markers TWIST, Akt2, and PI3Kα, as well as the stem cell marker ALDH1 were detectable, mainly in patient samples that also were positive for CTCs.
Using this threshold, 14 of 31 patient samples (45%) were considered positive for CTCs.
Green dotted line represents the healthy threshold of 1.32 CTCs/mL above which patients are defined as positive for CTCs.
Of 27 patients with recurrent disease, 14 were positive for CTCs and 13 of the CTC-positive patients were also positive for CD44-positive CTCs.
Samples from fourteen of these patients were positive for at least two of the chosen markers and were therefore considered positive for CTCs.
In this study three patients, who were not positive for CTCs prior to surgery, tested positive for CTCs after surgery.
Twelve patients initially positive for CTCs and positive for at least one of the EMT markers or ALDH1 or both were further tested for CD34 to exclude potential interference of normal hematopoietic stem cells in the blood samples.
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