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

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 me...

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Autores principales: Kim, Dae-Dong, Yang, Chun-Seok, Chae, Hyun-Dong, Kwak, Sang-Gyu, Jeon, Chang-Ho
Formato: Online Artículo Texto
Lenguaje:English
Publicado: D.A. Spandidos 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5494678/
https://www.ncbi.nlm.nih.gov/pubmed/28693240
http://dx.doi.org/10.3892/ol.2017.6226
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author Kim, Dae-Dong
Yang, Chun-Seok
Chae, Hyun-Dong
Kwak, Sang-Gyu
Jeon, Chang-Ho
author_facet Kim, Dae-Dong
Yang, Chun-Seok
Chae, Hyun-Dong
Kwak, Sang-Gyu
Jeon, Chang-Ho
author_sort Kim, Dae-Dong
collection PubMed
description 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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spelling pubmed-54946782017-07-07 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 Oncol Lett Articles 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. D.A. Spandidos 2017-07 2017-05-22 /pmc/articles/PMC5494678/ /pubmed/28693240 http://dx.doi.org/10.3892/ol.2017.6226 Text en Copyright: © Kim et al. This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License (https://creativecommons.org/licenses/by-nc-nd/4.0/) , which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.
spellingShingle Articles
Kim, Dae-Dong
Yang, Chun-Seok
Chae, Hyun-Dong
Kwak, Sang-Gyu
Jeon, Chang-Ho
Melanoma antigen-encoding gene family member A1-6 and hTERT in the detection of circulating tumor cells following CD45(−) depletion and RNA extraction
title Melanoma antigen-encoding gene family member A1-6 and hTERT in the detection of circulating tumor cells following CD45(−) depletion and RNA extraction
title_full Melanoma antigen-encoding gene family member A1-6 and hTERT in the detection of circulating tumor cells following CD45(−) depletion and RNA extraction
title_fullStr Melanoma antigen-encoding gene family member A1-6 and hTERT in the detection of circulating tumor cells following CD45(−) depletion and RNA extraction
title_full_unstemmed Melanoma antigen-encoding gene family member A1-6 and hTERT in the detection of circulating tumor cells following CD45(−) depletion and RNA extraction
title_short Melanoma antigen-encoding gene family member A1-6 and hTERT in the detection of circulating tumor cells following CD45(−) depletion and RNA extraction
title_sort melanoma antigen-encoding gene family member a1-6 and htert in the detection of circulating tumor cells following cd45(−) depletion and rna extraction
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5494678/
https://www.ncbi.nlm.nih.gov/pubmed/28693240
http://dx.doi.org/10.3892/ol.2017.6226
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