Circulating tumor cells in different stages of colorectal cancer
Abstract
Introduction. Liquid biopsies are noninvasive tests using blood or body fluids to detect circulating tumor cells (CTCs) or the products of tumor cells, such as fragments of nucleic acids or proteins that are shed into biological fluids from primary tumor or its metastates. The analysis of published clinical studies provides coherent evidence that the presence of CTCs detected in peripheral blood is a strong prognostic factor in patients with colorectal carcinoma (CRC). The aim of the study was to implement size-based separation protocol of CTCs in CRC patients.
Material and methods. Patients diagnosed with different stages of CRC (n = 98) were included in the study. All patients have been diagnosed for colorectal adenocarcinoma by pathology examination, 45 patients with colon carcinoma and 53 with rectosigmoid cancer. A size-based separation method (MetaCell®) for viable CTC enrichment from peripheral blood was used to assess the presence of CTCs by cytomorphological evaluation using vital fluorescence microscopy.
Results. Cytomorphological analysis revealed that 81 (83%) tested samples were CTC-positive and 17 (17%) were CTC-negative. We report a successful isolation of CTCs with proliferation potential in patients with CRC. The CTCs were cultured in vitro for further downstream applications. Some of the isolated CTCs were able to grow in vitro for 6 months as a standard cell culture.
Conclusions. We established a reliable, inexpensive and relatively fast protocol for CTCs enrichment in CRC patients by means of vital fluorescence staining which enables their further analysis in vitro.
Keywords: circulating tumor cellscolon cancerrectosigmoid cancerstagingcell cultureMetaCell®
References
- Rahbari NN, Aigner M, Thorlund K, et al. Meta-analysis shows that detection of circulating tumor cells indicates poor prognosis in patients with colorectal cancer. Gastroenterology. 2010; 138(5): 1714–1726.
- Liberko M, Kolostova K, Bobek V. Essentials of circulating tumor cells for clinical research and practice. Crit Rev Oncol Hematol. 2013; 88(2): 338–356.
- Eliasova P, Kolostova K, Kobierzycki C, et al. Clinical studies monitoring circulating and disseminated tumor cells in gastrointestinal cancers. Folia Histochem Cytobiol. 2013; 51(4): 265–277.
- Huang MY, Tsai HL, Huang JJ, et al. Clinical Implications and Future Perspectives of Circulating Tumor Cells and Biomarkers in Clinical Outcomes of Colorectal Cancer. Transl Oncol. 2016; 9(4): 340–347.
- Tan C, Zhou L, El-Deiry W. Circulating tumor cells versus circulating tumor DNA in colorectal cancer: pros and cons. Curr Colorectal Cancer Rep. 2016; 12(3): 151–161.
- Kolostova K, Matkowski R, Jędryka M, et al. The added value of circulating tumor cells examination in ovarian cancer staging. Am J Cancer Res. 2015; 5(11): 3363–3375.
- Kolostova K, Spicka J, Matkowski R, et al. Isolation, primary culture, morphological and molecular characterization of circulating tumor cells in gynecological cancers. Am J Transl Res. 2015; 7(7): 1203–1213.
- Cegan M, Kolostova K, Matkowski R, et al. In vitro culturing of viable circulating tumor cells of urinary bladder cancer. Int J Clin Exp Pathol. 2014; 7(10): 7164–7171.
- Bobek V, Matkowski R, Gürlich R, et al. Cultivation of circulating tumor cells in esophageal cancer. Folia Histochem Cytobiol. 2014; 52(3): 171–177.
- Kolostova K, Zhang Y, Hoffman RM, et al. In vitro culture and characterization of human lung cancer circulating tumor cells isolated by size exclusion from an orthotopic nude-mouse model expressing red fluorescent protein. J Fluoresc. 2014; 24(5): 1531–1536.
- Hermanek P. pTNM and residual tumor classifications: problems of assessment and prognostic significance. World J Surg. 1995; 19(2): 184–190.
- Cohen SJ, Punt CJA, Iannotti N, et al. Relationship of circulating tumor cells to tumor response, progression-free survival, and overall survival in patients with metastatic colorectal cancer. J Clin Oncol. 2008; 26(19): 3213–3221.
- Cohen SJ, Punt CJA, Iannotti N, et al. Prognostic significance of circulating tumor cells in patients with metastatic colorectal cancer. Ann Oncol. 2009; 20(7): 1223–1229.
- Groot Koerkamp B, Rahbari NN, Büchler MW, et al. Circulating tumor cells and prognosis of patients with resectable colorectal liver metastases or widespread metastatic colorectal cancer: a meta-analysis. Ann Surg Oncol. 2013; 20(7): 2156–2165.
- Matsusaka S, Suenaga M, Mishima Y, et al. Circulating tumor cells as a surrogate marker for determining response to chemotherapy in Japanese patients with metastatic colorectal cancer. Cancer Sci. 2011; 102(6): 1188–1192.
- Schuster R, Max N, Mann B, et al. Quantitative real-time RT-PCR for detection of disseminated tumor cells in peripheral blood of patients with colorectal cancer using different mRNA markers. Int J Cancer. 2004; 108(2): 219–227.
- Sher YP, Shih JY, Yang PC, et al. Prognosis of non-small cell lung cancer patients by detecting circulating cancer cells in the peripheral blood with multiple marker genes. Clin Cancer Res. 2005; 11(1): 173–179.
- Oberg ANV, Lindmark GE, Israelsson ACE, et al. Detection of occult tumour cells in lymph nodes of colorectal cancer patients using real-time quantitative RT-PCR for CEA and CK20 mRNAS. Int J Cancer. 2004; 111(1): 101–110.
- Ghajar C, Bissell M. Metastasis: Pathways of parallel progression. Nature. 2016; 540(7634): 528–529.
- Hosseini H, Obradović M, Hoffmann M, et al. Early dissemination seeds metastasis in breast cancer. Nature. 2016; 540(7634): 552–558.
- Yen LC, Yeh YS, Chen CW, et al. Detection of KRAS oncogene in peripheral blood as a predictor of the response to cetuximab plus chemotherapy in patients with metastatic colorectal cancer. Clin Cancer Res. 2009; 15(13): 4508–4513.
![](https://journals.viamedica.pl/plugins/generic/popups/images/icons/close.png)