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Primary cultures of human colon cancer as a model to study cancer stem cells

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Tumor Biology

Abstract

The principal cause of death in cancer involves tumor progression and metastasis. Since only a small proportion of the primary tumor cells, cancer stem cells (CSCs), which are the most aggressive, have the capacity to metastasize and display properties of stem cells, it is imperative to characterize the gene expression of diagnostic markers and to evaluate the drug sensitivity in the CSCs themselves. Here, we have examined the key genes that are involved in the progression of colorectal cancer and are expressed in cancer stem cells. Primary cultures of colorectal cancer cells from a patient’s tumors were studied using the flow cytometry and cytological methods. We have evaluated the clinical and stem cell marker expression in these cells, their resistance to 5-fluorouracil and irinotecan, and the ability of cells to form tumors in mice. The data shows the role of stem cell marker Oct4 in the resistance of primary colorectal cancer tumor cells to 5-fluorouracil.

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References

  1. Siegel RL, Miller KD, Jemal A. Cancer statistics, 2016. CA Cancer J Clin. 66(1):7–30.

  2. Chang GJ, CY H, Eng C, Skibber JM, Rodriguez-Bigas MA. Practical application of a calculator for conditional survival in colon cancer. J Clin Oncol. 2009;27:5938–43.

    Article  PubMed  Google Scholar 

  3. Ishida H, Fujita K, Akiyama Y, Sunakawa Y, Yamashita K, Mizuno K, et al. Regimen selection for first-line FOLFIRI and FOLFOX based on UGT1A1 genotype and physical background is feasible in Japanese patients with advanced colorectal cancer. Jpn J Clin Oncol. 2011;41:617–23.

    Article  PubMed  Google Scholar 

  4. Sanli UA, Karabulut B, Uslu R, Korkut M, Goker E. Single-agent irinotecan for recurrent/metastatic colorectal cancer: a retrospective analysis. Med Princ Pract. 2006;15:288–92.

    Article  PubMed  Google Scholar 

  5. Valent P, Bonnet D, De Maria R, Lapidot T, Copland M, Melo JV, et al. Cancer stem cell definitions and terminology: the devil is in the details. Nat Rev Cancer. 2012;12:767–75.

    Article  CAS  PubMed  Google Scholar 

  6. Lapidot T, Sirard C, Vormoor J, Murdoch B, Hoang T, Caceres-Cortes J, et al. A cell initiating human acute myeloid leukaemia after transplantation into SCID mice. Nature. 1994;367:645–8.

    Article  CAS  PubMed  Google Scholar 

  7. Ricci-Vitiani L, Fabrizi E, Palio E, De Maria R. Colon cancer stem cells. J Mol Med (Berl). 2009;87:1097–104.

    Article  Google Scholar 

  8. Fabian A, Barok M, Vereb G, Szollosi J. Die hard: are cancer stem cells the Bruce Willises of tumor biology? Cytometry A. 2009;75:67–74.

    Article  PubMed  Google Scholar 

  9. Vaiopoulos AG, Kostakis ID, Koutsilieris M, Papavassiliou AG. Colorectal cancer stem cells. Stem Cells. 2012;30:363–71.

    Article  CAS  PubMed  Google Scholar 

  10. Mathonnet M, Perraud A, Christou N, Akil H, Melin C, Denizot Y, et al. Hallmarks in colorectal cancer: angiogenesis and cancer stem-like cells. World J Gastroenterol. 2014;20:4189–96.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  11. O’Brien CA, Pollett A, Gallinger S, Dick JE. A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature. 2007;445:106–10.

    Article  PubMed  Google Scholar 

  12. Van de Wetering M, Sancho E, Verweij C, de Lau W, Oving I, Hurlstone A, van der Horn K, Clevers H. The beta-catenin/TCF-4 complex imposes a crypt progenitor phenotype on colorectal cancer cells. Cell. 2002;111:241–50.

    Article  CAS  PubMed  Google Scholar 

  13. XS W, Xi HQ, Chen L. Lgr5 is a potential marker of colorectal carcinoma stem cells that correlates with patient survival. World J Surg Oncol. 2012;10:244.14.

    Google Scholar 

  14. Ritsma L, Ellenbroek SI, Zomer A, Snippert HJ, de Sauvage FJ, van Rheenen J, et al. Intestinal crypt homeostasis revealed at single-stem-cell level by in vivo live imaging. Nature. 2014;507:362–5.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Gargett CE, Schwab KE, Zillwood RM, Nguyen HP, Wu D. Isolation and culture of epithelial progenitors and mesenchymal stem cells from human endometrium. Biol Reprod. 2009;80:1136–45.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Ebben JD, Treisman DM, Zorniak M, Kutty RG, Clark PA, Kuo JS. The cancer stem cell paradigm: a new understanding of tumor development and treatment. Expert Opin Ther Targets. 2010;14:621–32.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. CS Y, Huang AC, Lai KC, Huang YP, Lin MW, Chung JG, et al. Diallyltrisulfide induces apoptosis in human primary colorectal cancer cells. Oncol Rep. 2012;28:94954.

    Google Scholar 

  18. Todaro M, Orlando V, Cicero G, Caccamo N, Meraviglia S, Dieli F, et al. Chemotherapy sensitizes colon cancer initiating cells to Vy9V52 T cell-mediated cytotoxicity. PLoS One. 2013;8.

  19. Khan IN, Al-Karim S, Bora RS, Chaudhary AG, Saini KS. Cancer stem cells: a challenging paradigm for designing targeted drug therapies. Drug Discov Today. 2015;20(10):1205–16.

    Article  CAS  PubMed  Google Scholar 

  20. Tomayko MM, Reynolds CP, Euhus DM, Hudd C, LaRegina MC, Johnson FE. Determination of subcutaneous tumor size in athymic (nude) mice. Cancer Chemother Pharmacol. 1989;24(3):148–54.

    Article  CAS  PubMed  Google Scholar 

  21. Liskovykh MA, Chuikin IA, Ranjan A, Safina DA, Tolkunova EN, Tomilin AN, et al. Generation of rat induced pluripotent stem cells: the analysis of reprogramming and culturing media. Tsitologiia. 2011;53:939–45.

    CAS  PubMed  Google Scholar 

  22. Davydov-Sinitsyn AP, Bazhenova OV, Liskovykh MA, Ponomartsev CV, Tomilin AN, Tolkunova EN, et al. In vitro derivation and characterization of a colorectal cancer stem cell subpopulation. Tsitologiia. 2013;55:318–23.

    CAS  PubMed  Google Scholar 

  23. Wolmark N, Rockette H, Fisher B, Wickerham DL, Redmond C, Petrelli NJ, et al. The benefit of leucovorin-modulated fluorouracil as postoperative adjuvant therapy for primary colon cancer: results from National Surgical Adjuvant Breast and Bowel Project protocol C-03. J ClinOncol. 1993;11:187987.

    Google Scholar 

  24. Cunningham D, Pyrhonen S, James RD, Punt CJ, Hickish TF, Herait P, et al. Randomised trial of irinotecan plus supportive care versus supportive care alone after fluorouracil failure for patients with metastatic colorectal cancer. Lancet. 1998;352:1413–8.

    Article  CAS  PubMed  Google Scholar 

  25. Hochedlinger K, Yamada Y, Beard C, Jaenisch R. Ectopic expression of Oct-4 blocks progenitor-cell differentiation and causes dysplasia in epithelial tissues. Cell. 2005;121:465–77.

    Article  CAS  PubMed  Google Scholar 

  26. DeSantis CE, Lin CC, Mariotto AB, Siegel RL, Stein KD, Jemal A. Cancer treatment and survivorship statistics, 2014. CA Cancer J Clin. 2014;64:252–71.

    Article  PubMed  Google Scholar 

  27. Siegel R, DeSantis C, Jemal A. Colorectal cancer statistics, 2014. CA Cancer J Clin. 2014;64:104–17.

    Article  PubMed  Google Scholar 

  28. Gungor C, Hofmann BT, Wolters-Eisfeld G, Bockhorn M. Pancreatic cancer. Br J Pharmacol. 2014;171:849–58.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Kaddis N, Saif MW. Second-line treatment for pancreatic cancer. JOP. 2014;15:344347.

    Google Scholar 

  30. Tanase CP, Neagu AI, Necula LG, Mambet C, Enciu AM, Albulescu R, et al. Cancer stem cells: involvement in pancreatic cancer pathogenesis and perspectives on cancer therapeutics. World J Gastroenterol. 2014;20:10790–801.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Muqbil I, Bao GW, El-Kharraj R, Shah M, Mohammad RM, Azmi AS et al. Systems and network pharmacology approaches to cancer stem cells research and therapy. J Stem Cell Res Ther. 2012;suppl 7.

  32. Wang YJ, Herlyn M. The emerging roles of Oct4 in tumor-initiating cells. Am J Phys Cell Phys. 2015;309(11):C709–18.

    CAS  Google Scholar 

  33. Poursani EM, Mohammad Soltani B, Mowla SJ. Differential expression of OCT4 pseudogenes in pluripotent and tumor cell lines. Cell J. 2016;18(1):28–36.

    PubMed  PubMed Central  Google Scholar 

  34. Kobayashi I, Takahashi F, Nurwidya F, Nara T, Hashimoto M, Takahashi K. Oct4 plays a crucial role in the maintenance of gefitinib-resistant lung cancer stem cells. Biochem Biophys Res Commun. 2016;473(1):125–32.

    Article  CAS  PubMed  Google Scholar 

  35. Fazlul H. Sarkar. Novel holistic approaches for overcoming therapy resistance in pancreatic and colon cancers. Med Princ Pract 2015.

  36. Meacham CE, Morrison SJ. Tumour heterogeneity and cancer cell plasticity. Nature. 2013;501:328–37.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  37. Todaro M, Alea MP, Di Stefano AB, Cammareri P, Vermeu-len L, Stassi G. Colon cancer stem cells dictate tumor growth and resist cell death by production of interleukin-4. Cell Stem Cell. 2007;1:389–402.

    Article  CAS  PubMed  Google Scholar 

  38. Dallas NA, Xia L, Fan F, Gray MJ, Gaur P, Ellis LM, et al. Chemoresistant colorectal cancer cells, the cancer stem cell phenotype, and increased sensitivity to insulin-like growth factor-I receptor inhibition. Cancer Res. 2009;69:1951–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  39. Dieter SM, Ball CR, Hoffmann CM, Nowrouzi A, Herbst F, Glimm H, et al. Distinct types of tumor-initiating cells form human colon cancer tumors and metastases. Cell Stem Cell. 2011;9:357–65.

    Article  CAS  PubMed  Google Scholar 

  40. Pointer KB, Clark PA, Zorniak M, Alrfaei BM, Kuo JS. Glioblastoma cancer stem cells: biomarker and therapeutic advances. Neurochem Int. 2014;71:1–7.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Pitynski K, Banas T, Pietrus M, Milian-Ciesielska K, Ludwin A, Okon K. SOX-2, but not Oct4, is highly expressed in early-stage endometrial adenocarcinoma and is related to tumour grading. Int J Clin Exp Pathol. 2015;8:8189–98.

    PubMed  PubMed Central  Google Scholar 

  42. Ji J, Wei X, Wang Y. Embryonic stem cell markers Sox-2 and OCT4 expression and their correlation with WNT signal pathway in cervical squamous cell carcinoma. Int J Clin Exp Pathol. 2014;7:2470–86.

    PubMed  PubMed Central  Google Scholar 

  43. Yang F, Gao Y, Geng J, Qu D, Han Q, Chen G, et al. Elevated expression of SOX2 and FGFR1 in correlation with poor prognosis in patients with small cell lung cancer. Int J Clin Exp Pathol. 2013;6:2846–54.

    PubMed  PubMed Central  Google Scholar 

  44. Wang Q, He W, Lu C, Wang Z, Wang J, Suo Z, et al. Oct3/4 and Sox2 are significantly associated with an unfavorable clinical outcome in human esophageal squamous cell carcinoma. Anticancer Res. 2009;29:1233–41.

    CAS  PubMed  Google Scholar 

  45. Saigusa S, Tanaka K, Toiyama Y, Yokoe T, Okugawa, Kusunoki M, et al. Correlation of CD133, OCT4, and SOX2 in rectal cancer and their association with distant recurrence after chemoradiotherapy. Ann Surg Oncol. 2009;16:3488–98.

    Article  PubMed  Google Scholar 

  46. Hosokawa Y, Takahashi H, Inoue A, Kawabe Y, Funahashi Y, Tanaka J, et al. Oct-3/4 modulates the drug- resistant phenotype of glioblastoma cells through expression of ATP binding cassette transporter G2. Biochim Biophys Acta. 2015;1850:1197–205.

    Article  CAS  PubMed  Google Scholar 

  47. Kashihara H, Shimada M, Kurita N, Iwata T, Sato H, Matsumoto N, et al. CD133 expression is correlated with poor prognosis in colorectal cancer. Hepato-Gastroenterology. 2014;61:1563–7.

    PubMed  Google Scholar 

  48. Vicente-Duenas C, Gutierrez de Diego J, Rodriguez FD, Jimenez R, Cobaleda C. The role of cellular plasticity in cancer development. Curr Med Chem. 2009;16:3676–85.

    Article  CAS  PubMed  Google Scholar 

  49. Ren F, Sheng WQ, Du X. CD133: a cancer stem cells marker, is used in colorectal cancers. World J Gastroenterol. 2013;19:2603–11.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  50. Deng YH, XX P, Huang MJ, Xiao J, Zhou JM, Lin EH, et al. 5-Fluorouracil upregulates the activity of Wnt signaling pathway in CD133-positive colon cancer stemlike cells. Chin J Cancer. 2010;29:810–5.

    Article  CAS  PubMed  Google Scholar 

  51. Qi L, Sun B, Liu Z, Li H, Gao J, Leng X. Dickkopf-1 inhibits epithelial-mesenchymal transition of colon cancer cells and contributes to colon cancer suppression. Cancer Sci. 2012;103:828–35.

    Article  CAS  PubMed  Google Scholar 

  52. Fan F, Bellister S, Lu J, Ye X, Boulbes DR, Ellis LM, et al. The requirement for freshly isolated human colorectal cancer (CRC) cells in isolating CRC stem cells. Br J Cancer. 2015;112:539–46.

    Article  CAS  PubMed  Google Scholar 

  53. Barker N, Clevers H. Leucine-rich repeat-containing G-protein-coupled receptors as markers of adult stem cells. Gastroenterology. 2010;138:1681–96.

    Article  CAS  PubMed  Google Scholar 

  54. Sangiorgi E, Capecchi MR. Bmi1 is expressed in vivo in intestinal stem cells. Nat Genet. 2008;40:915–20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Authors and Affiliations

  1. Institute of Cytology, Russian Academy of Sciences, Tikhoretsky ave. 4, St. Petersburg, Russia, 194064

    Sergey Koshkin, Nikolai Petrov, Alexey Tomilin & Elena Tolkunova

  2. N.N. Petrov Oncology Research Institute, St. Petersburg, 197758, Russia

    Anna Danilova

  3. Russian Research Centre for Radiology and Surgical Technologies, St. Petersburg, 197758, Russia

    Grigory Raskin

  4. Theodosius Dobzhansky Center for Genome Bioinformatics, St. Petersburg State University, St. Petersburg, 199034, Russia

    Olga Bajenova & Stephen J. O’Brien

  5. Department of Genetics and Biotechnology, St. Petersburg State University, St. Petersburg, 199034, Russia

    Olga Bajenova

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  1. Sergey Koshkin
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  2. Anna Danilova
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  3. Grigory Raskin
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  4. Nikolai Petrov
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  5. Olga Bajenova
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  6. Stephen J. O’Brien
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  7. Alexey Tomilin
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  8. Elena Tolkunova
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Correspondence to Sergey Koshkin.

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Grant support

This work was financially supported by the Russian Science Foundation (project 14-50-00068) to the Molecular Stem Cell Biology Laboratory, Institute of Cytology RAS, and by the Government of the Russian Federation mega grant 11G34.31.0068 to Dr. S. J. O’Brien.

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Koshkin, S., Danilova, A., Raskin, G. et al. Primary cultures of human colon cancer as a model to study cancer stem cells. Tumor Biol. 37, 12833–12842 (2016). https://doi.org/10.1007/s13277-016-5214-8

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  • DOI: https://doi.org/10.1007/s13277-016-5214-8

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