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. 2014 Jun;7(6):2192-2198.
doi: 10.3892/ol.2014.2048. Epub 2014 Apr 8.

A rapid nested polymerase chain reaction method to detect circulating cancer cells in breast cancer patients using multiple marker genes

Lei Liu  1 , Chunhu Ma  2 , Qian Xu  3 , Luyang Cheng  1 , Lijun Xiao  1 , Dawei Xu  1 , Yaxian Gao  1 , Jianping Wang  1 , Hongru Song  1
Affiliations

A rapid nested polymerase chain reaction method to detect circulating cancer cells in breast cancer patients using multiple marker genes

Lei Liu et al. Oncol Lett. 2014 Jun.

Abstract

The aim of the present study was to develop a simple and rapid method for the detection of circulating cancer cells using multiple tumor markers and to investigate the clinical significance of circulating cancer cells in breast cancer patients. A novel rapid nested polymerase chain reaction (PCR) assay, with high sensitivity and specificity, was evaluated, which was considered to be suitable for clinical application. The rapid nested PCR method was used to detect the circulating cancer cells of 142 breast cancer patients, using a panel of marker genes (FAM83A, NPY1R and KRT19), which were identified by the Digital Gene Expression Displayer Tool of the National Cancer Institute-Cancer Genome Anatomy Project. In total, 79.6% of the 142 breast cancer patient blood samples were found to express at least one tumor marker. In addition, the number of positive markers was found to significantly correlate with the disease stage and presence of distant metastasis. Furthermore, positivity for more than one tumor marker appeared to predict a reduced survival time in breast cancer patients.

Keywords: breast cancer; circulating cancer cells; nested polymerase chain reaction; tumor marker.

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Figures

Figure 1
Figure 1
Evaluation of the rapid nested polymerase chain reaction (PCR). (A) The same detection rate was achieved using the (a) novel rapid and the (b) traditional nested PCR assays. Lanes 1–15, breast cancer patients; lane 16, healthy donor; and lane 17, negative control. (B) Sensitivity of the (a) novel rapid and the (b) traditional nested PCR assays. Lanes 1–7, 10-fold serial dilution of MCF-7 using peripheral blood mononuclear cells obtained from healthy donors (Lane 1, 102; lane, 2 103; lane 3, 104; lane 4, 105; lane 5, 106; lane 6, 107; and lane 7, 108 cells); and lane 8, negative control. (C) Amplification using outer and inner primers. Lanes 1 and 2, outer primers only; lanes 3 and 4, outer and inner primers; and lanes 5 and 6, inner primers only.
Figure 2
Figure 2
Analysis of positive detection rates with a multimarker gene panel. The positive detection rate for (A) the three individual marker genes. (B) The positive detection rate increased with the panel of markers.
Figure 3
Figure 3
Kaplan-Meier survival analysis for breast cancer patients, grouped according to the number of tumor markers detected in the peripheral blood.
See this image and copyright information in PMC

References

    1. Gardner B, Feldman J. Are positive axillary nodes in breast cancer markers for incurable disease? Ann Surg. 1993;218:270–278. - PMC - PubMed
    1. Zhao S, Yang H, Zhang M, et al. Circulating tumor cells (CTCs) detected by triple-marker EpCAM, CK19, and hMAM RT-PCR and their relation to clinical outcome in metastatic breast cancer patients. Cell Biochem Biophys. 2013;65:263–273. - PubMed
    1. Reinholz MM, Nibbe A, Jonart LM, et al. Evaluation of a panel of tumor markers for molecular detection of circulating cancer cells in women with suspected breast cancer. Clin Cancer Res. 2005;11:3722–3732. - PubMed
    1. Margossian A, Diaz J, Corvalan A. In silico analysis of breast cancer transcriptome libraries distinguish tumor subclasses. Cancer Res. 2009;69(Suppl 3):S1165.
    1. Rahbari NN, Bork U, Motschall E, et al. Molecular detection of tumor cells in regional lymph nodes is associated with disease recurrence and poor survival in node-negative colorectal cancer: a systematic review and meta-analysis. J Clin Oncol. 2012;30:60–70. - PubMed

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