Detection of circulating tumor cells by reverse transcription‑quantitative polymerase chain reaction and magnetic activated cell sorting in the peripheral blood of patients with hepatocellular carcinoma

Guo,Limin; Xu,Donghai; Lu,Yan; Peng,Jirun; Jiang,Li

doi:10.3892/mmr.2017.7372

November-2017 Volume 16 Issue 5

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November-2017 Volume 16 Issue 5

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Detection of circulating tumor cells by reverse transcription‑quantitative polymerase chain reaction and magnetic activated cell sorting in the peripheral blood of patients with hepatocellular carcinoma

Authors:
- Limin Guo
- Donghai Xu
- Yan Lu
- Jirun Peng
- Li Jiang
View Affiliations / Copyright

Affiliations: Department of Hepatobiliary Surgery, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, P.R. China, Department of Radiology, Beijing Ditan Hospital, Capital Medical University, Beijing 100015, P.R. China, Department of Surgery, Beijing Shijitan Hospital, Capital Medical University, Beijing 100038, P.R. China

Copyright: © Guo et al. This is an open access article distributed under the terms of Creative Commons Attribution License.
Pages: 5894-5900
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Published online on: August 28, 2017

https://doi.org/10.3892/mmr.2017.7372
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Abstract

Hepatocellular carcinoma (HCC) is one of the most lethal malignancies worldwide. Circulating tumor cells (CTCs) are considered a major cause of recurrence and metastasis in cancer; however, the detection of CTCs is challenging owing to their very low numbers in peripheral blood (around 10 CTCs per 1,000,000 erythrocytes). Cancer‑testis antigens (CTAs) are specific tumor markers for CTCs. The present study aimed to evaluate the sensitivity and specificity of reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR) for the detection of nine CTAs as well as placenta‑specific antigen 1 (PLAC1) in peripheral blood mononuclear cell (PBMC) samples collected from 51 patients with HCC. The effectiveness of magnetic‑activated cell sorting (MACS) for tumor‑cell enrichment, through the depletion of CD45+ leukocytes in PBMC samples, was also assessed. Immunocytochemistry along with hematoxylin and eosin staining demonstrated that RT‑qPCR achieved an overall positive detection rate for CTAs and PLAC1 of 70.6%; the highest rates were observed for melanoma‑associated antigen A3 (MAGEA3), synovial sarcoma X breakpoint 1, MAGEA1, NY‑ESO‑1, L antigen 1 and PLAC1. MACS‑detected intact CTCs in PBMCs were confirmed by H&E staining and morphological assessment; 12 out of 19 (63.2%) patients were identified as positive for CTAs. Screening for these five CTAs and PLAC1 by RT‑qPCR may offer a potentially valuable prognostic tool with good sensitivity and specificity in patients with HCC that may be enhanced by MACS.

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1	Parkin DM, Bray F, Ferlay J and Pisani P: Global cancer statistics, 2002. CA Cancer J Clin. 55:74–108. 2005. View Article : Google Scholar : PubMed/NCBI
2	Parkin DM: Global cancer statistics in the year 2000. Lancet Oncol. 2:533–543. 2001. View Article : Google Scholar : PubMed/NCBI
3	Elshimali YI and Grody WW: The clinical significance of circulating tumor cells in the peripheral blood. Diagn Mol Pathol. 15:187–194. 2006. View Article : Google Scholar : PubMed/NCBI
4	Hosokawa M, Hayata T, Fukuda Y, Arakaki A, Yoshino T, Tanaka T and Matsunaga T: Size-selective microcavity array for rapid and efficient detection of circulating tumor cells. Anal Chem. 82:6629–6635. 2010. View Article : Google Scholar : PubMed/NCBI
5	Samija I, Lukac J, Kusić Z, Situm M and Samija M: Detection of disseminated melanoma cells by reverse-transcription-polymerase chain reaction. Coll Antropol. 31:1187–1194. 2007.PubMed/NCBI
6	Schefe JH, Lehmann KE, Buschmann IR, Unger T and Funke-Kaiser H: Quantitative real-time RT-PCR data analysis: Current concepts and the novel ‘gene expression's CT difference’ formula. J Mol Med (Berl). 84:901–910. 2006. View Article : Google Scholar : PubMed/NCBI
7	Morimoto O, Nagano H, Miyamoto A, Fujiwara Y, Kondo M, Yamamoto T, Ota H, Nakamura M, Wada H, Damdinsuren B, et al: Association between recurrence of hepatocellular carcinoma and alpha-fetoprotein messenger RNA levels in peripheral blood. Surg Today. 35:1033–1041. 2005. View Article : Google Scholar : PubMed/NCBI
8	Yao F, Guo JM, Xu CF, Lou YL, Xiao BX, Zhou WH, Chen J, Hu YR, Liu Z and Hong GF: Detecting AFP mRNA in peripheral blood of the patients with hepatocellular carcinoma, liver cirrhosis and hepatitis. Clin Chim Acta. 361:119–127. 2005. View Article : Google Scholar : PubMed/NCBI
9	Lu Y, Wu LQ, Lü ZH, Wang XJ and Yang JY: Expression of SSX-1 and NY-ESO-1 mRNA in tumor tissues and its corresponding peripheral blood expression in patients with hepatocellular carcinoma. Chin Med J (Engl). 120:1042–1046. 2007.PubMed/NCBI
10	Mou DC, Cai SL, Peng JR, Wang Y, Chen HS, Pang XW, Leng XS and Chen WF: Evaluation of MAGE-1 and MAGE-3 as tumour-specific markers to detect blood dissemination of hepatocellular carcinoma cells. Br J Cancer. 86:110–116. 2002. View Article : Google Scholar : PubMed/NCBI
11	Zendman AJ, Ruiter DJ and Van Muijen GN: Cancer/testis-associated genes: Identification, expression profile, and putative function. J Cell Physiol. 194:272–288. 2003. View Article : Google Scholar : PubMed/NCBI
12	Peng JR, Chen HS, Mou DC, Cao J, Cong X, Qin LL, Wei L, Leng XS, Wang Y and Chen WF: Expression of cancer/testis (CT) antigens in Chinese hepatocellular carcinoma and its correlation with clinical parameters. Cancer Lett. 219:223–232. 2005. View Article : Google Scholar : PubMed/NCBI
13	Koslowski M, Sahin U, Mitnacht-Kraus R, Seitz G, Huber C and Türeci O: A placenta-specific gene ectopically activated in many human cancers is essentially involved in malignant cell processes. Cancer Res. 67:9528–9534. 2007. View Article : Google Scholar : PubMed/NCBI
14	Bustin SA: Absolute quantification of mRNA using real-time reverse transcription polymerase chain reaction assays. J Mol Endocrinol. 25:169–193. 2000. View Article : Google Scholar : PubMed/NCBI
15	Walzer N and Kulik LM: Hepatocellular carcinoma: Latest developments. Curr Opin Gastroenterol. 24:312–319. 2008. View Article : Google Scholar : PubMed/NCBI

Journals

International Journal of Molecular Medicine

International Journal of Oncology

Molecular Medicine Reports

Oncology Reports

Experimental and Therapeutic Medicine

Oncology Letters

Biomedical Reports

Molecular and Clinical Oncology

World Academy of Sciences Journal

International Journal of Functional Nutrition

International Journal of Epigenetics

Medicine International

Detection of circulating tumor cells by reverse transcription‑quantitative polymerase chain reaction and magnetic activated cell sorting in the peripheral blood of patients with hepatocellular carcinoma

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