Melanoma antigen-encoding gene family member A1-6 and hTERT in the detection of circulating tumor cells following CD45‑ depletion and RNA extraction

Kim,Dae‑Dong; Yang,Chun‑Seok; Chae,Hyun‑Dong; Kwak,Sang‑Gyu; Jeon,Chang‑Ho

doi:10.3892/ol.2017.6226

Melanoma antigen-encoding gene family member A1-6 and hTERT in the detection of circulating tumor cells following CD45‑ depletion and RNA extraction

Authors:
- Dae‑Dong Kim
- Chun‑Seok Yang
- Hyun‑Dong Chae
- Sang‑Gyu Kwak
- Chang‑Ho Jeon
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Affiliations: Department of Surgery, School of Medicine, Catholic University of Daegu, Daegu 42472, Republic of Korea, Department of Medical Statistics, School of Medicine, Catholic University of Daegu, Daegu 42472, Republic of Korea, Department of Laboratory Medicine, School of Medicine, Catholic University of Daegu, Daegu 42472, Republic of Korea
Published online on: May 22, 2017 https://doi.org/10.3892/ol.2017.6226
Pages: 837-843
Copyright: © Kim et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

A total of 76 blood samples from patients without malignant disease and 107 blood samples from patients with malignant disease were investigated for the presence of circulating tumor cells (CTCs). To detect CTCs, hematopoietic cells were removed from the blood samples and different RNA extraction methods were used to amplify the melanoma antigen‑encoding gene family member A1‑family member A6 (MAGE A1‑6) and the human telomerase reverse transcriptase (hTERT) gene as potential CTC markers. Comparison between four methods for extracting RNA from the blood was performed. The samples were enriched by cluster of differentiation 45 (CD45) antibody capturing, and the reverse transcription‑quantitative polymerase chain reaction was used to amplify the MAGE A1‑6 and hTERT genes. MAGE A1‑6 and hTERT gene expression levels were also evaluated in 14 cancer cell lines, and the MAGE A1‑6 and hTERT expression levels were 85.7 and 100%, respectively. The RNeasy method demonstrated the most sensitivity in the SNU1 cells mixed with blood, although the differences between methods was non‑significant. The positive expression levels of MAGE A1‑6 and hTERT was 11.8% in the control group and 58.9% in those with malignant disease. In the 70 patients with colorectal cancer, positive expression levels of MAGE A1‑6 or hTERT were significantly higher in stages T3 and T4 compared with in stages T1 and T2. The CTC detection method involving CD45 antibody capture, RNA extraction and MAGE A1‑6 and hTERT reverse transcription resulted in good rates of sensitivity and specificity. Thus, the present study concluded that MAGE A1‑6 and hTERT genes may be potential and practical markers for CTCs in a clinical setting.

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