Enrichment and characterization of cancer stem‑like cells from a cervical cancer cell line

Wang,Li; Guo,Huijie; Lin,Caiyu; Yang,Liuqi; Wang,Xiujie

doi:10.3892/mmr.2014.2063

Enrichment and characterization of cancer stem‑like cells from a cervical cancer cell line

Authors:
- Li Wang
- Huijie Guo
- Caiyu Lin
- Liuqi Yang
- Xiujie Wang
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Affiliations: Laboratory of Experimental Oncology, State Key Laboratory of Biotherapy, West China Hospital, West China Clinical Medical School, Sichuan University, Chengdu, Sichuan 610041, P.R. China
Published online on: March 20, 2014 https://doi.org/10.3892/mmr.2014.2063
Pages: 2117-2123
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

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Abstract

Cancer stem cells (CSCs) are proposed to be responsible for tumor recurrence, metastasis and the high mortality rate of cancer patients. Isolation and identification of CSCs is crucial for basic and preclinical studies. However, as there are currently no universal markers for the isolation and identification of CSCs in any type of cancer, the method for isolating CSCs from primary cancer tissues or cell lines is costly and ineffective. In order to establish a reliable model of cervical cancer stem cells for basic and preclinical studies, the present study was designed to enrich cervical cancer CSCs using a nonadhesive culture system and to characterize their partial stemness phenotypes. Human cervical cancer cells (HeLa) were cultured using a nonadhesive culture system to generate tumor spheres. Their stemness characteristics were investigated through colony formation, tumor sphere formation, self‑renewal, toluidine blue staining, chemoresistance, invasion assays, reverse transcription‑polymerase chain reaction, immunofluorescence staining of putative stem cell markers, including octamer‑binding transcription factor 4, SRY‑box 2 and aldehyde dehydrogenase 1 family, member A1, and adipogenic differentiation induction. Typical tumor spheres were formed within 5‑7 days under this nonadhesive culture system. Compared with the adherent parental HeLa cells, the colony formation capacity, self‑renewal potential, light cell population, cell invasion, chemoresistance and expression of putative stem cell markers of the tumor sphere cells increased significantly, and a subpopulation of tumor sphere cells were induced into adipogenic differentiation. Using the nonadhesive culture system, a reliable model of cervical cancer stem cells was established, which is inexpensive, effective and simple compared with the ultra‑low attachment serum free culture method. The stemness characteristics of the tumor sphere HeLa cells mirrored the CSC phenotypes. This CSC model may be useful for basic and preclinical studies of cervical cancer and other types of cancer.

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1	Jemal A, Bray F, Center MM, et al: Global Cancer Statistics. CA Cancer J Clin. 61:69–90. 2011. View Article : Google Scholar
2	Lea JS and Lin KY: Cervical Cancer. Obstet Gynecol Clin North Am. 39:233–253. 2012. View Article : Google Scholar
3	Green JA, Kirwan JM, Tierney JF, et al: Survival and recurrence after concomitant chemotherapy and radiotherapy for cancer of the uterine cervix: a systematic review and meta-analysis. Lancet. 358:781–786. 2001. View Article : Google Scholar : PubMed/NCBI
4	Pectasides D, Kamposioras K, Papaxoinis G, et al: Chemotherapy for recurrent cervical cancer. Cancer Treat Rev. 34:603–613. 2008. View Article : Google Scholar : PubMed/NCBI
5	Ichim CV and Wells RA: First among equals: the cancer cell hierarchy. Leuk Lymphoma. 47:2017–2027. 2006. View Article : Google Scholar : PubMed/NCBI
6	Mao XG, Guo G, Wang P, et al: Maintenance of critical properties of brain tumor stem-like cells after cryopreservation. Cell Mol Neurobiol. 30:775–786. 2010. View Article : Google Scholar : PubMed/NCBI
7	Cioce M, Gherardi S, Viglietto G, et al: Mammosphere-forming cells from breast cancer cell lines as a tool for the identification of CSC-like- and early progenitor-targeting drugs. Cell Cycle. 9:2878–2887. 2010. View Article : Google Scholar : PubMed/NCBI
8	Su YJ, Lai HM, Chang YW, et al: Direct reprogramming of stem cell properties in colon cancer cells by CD44. EMBO J. 30:3186–3199. 2011. View Article : Google Scholar : PubMed/NCBI
9	Zhang L, Jiao M, Li L, et al: Tumorspheres derived from prostate cancer cells possess chemoresistant and cancer stem cell properties. J Cancer Res Clin Oncol. 138:675–686. 2012. View Article : Google Scholar : PubMed/NCBI
10	López J, Poitevin A, Mendoza-Martínez V, et al: Cancer-initiating cells derived from established cervical cell lines exhibit stem-cell markers and increased radioresistance. BMC Cancer. 12:482012.PubMed/NCBI
11	Lee J, Kotliarova S, Kotliarov Y, et al: Tumor stem cells derived from glioblastomas cultured in bFGF and EGF more closely mirror the phenotype and genotype of primary tumors than do serum-cultured cell lines. Cancer Cell. 9:391–403. 2006. View Article : Google Scholar : PubMed/NCBI
12	Hueng DY, Sytwu HK, Huang SM, et al: Isolation and characterization of tumor stem-like cells from human meningiomas. J Neurooncol. 104:45–53. 2011. View Article : Google Scholar : PubMed/NCBI
13	Zhong Y, Guan K, Guo S, et al: Spheres derived from the human SK-RC-42 renal cell carcinoma cell line are enriched in cancer stem cells. Cancer Lett. 299:150–160. 2010. View Article : Google Scholar : PubMed/NCBI
14	Chen SF, Chang YC, Nieh S, et al: Nonadhesive culture system as a model of rapid sphere formation with cancer stem cell properties. PLoS One. 7:e318642012. View Article : Google Scholar : PubMed/NCBI
15	Ponti D, Costa A, Zaffaroni N, et al: Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties. Cancer Res. 65:5506–5511. 2005. View Article : Google Scholar : PubMed/NCBI
16	Wedel S, Hudak L, Seibel JM, et al: Molecular targeting of prostate cancer cells by a triple drug combination down-regulates integrin driven adhesion processes, delays cell cycle progression and interferes with the cdk-cyclin axis. BMC Cancer. 11:3752011. View Article : Google Scholar : PubMed/NCBI
17	Kim MJ, Kim YJ, Park HJ, et al: Apoptotic effect of red wine polyphenols on human colon cancer SNU-C4 cells. Food Chem Toxicol. 44:898–902. 2006. View Article : Google Scholar : PubMed/NCBI
18	Hu Y and Fu L: Targeting cancer stem cells: a new therapy to cure cancer patients. Am J Cancer Res. 2:340–356. 2012.PubMed/NCBI
19	Gilbert CA and Ross AH: Cancer stem cells: cell culture, markers, and targets for new therapies. J Cell Biochem. 108:1031–1038. 2009. View Article : Google Scholar : PubMed/NCBI
20	Kim RK, Kim MJ, Yoon CH, et al: A new 2-pyrone derivative, 5-bromo-3-(3-hydroxyprop-1-ynyl)-2H-pyran-2-one, suppresses stemness in glioma stem-like cells. Mol Pharmacol. 82:400–407. 2012. View Article : Google Scholar : PubMed/NCBI
21	Cheng W, Liu T, Wan X, et al: MicroRNA-199a targets CD44 to suppress the tumorigenicity and multidrug resistance of ovarian cancer-initiating cells. FEBS J. 279:2047–2059. 2012. View Article : Google Scholar : PubMed/NCBI
22	Mather JP: In vitro models. Stem Cells. 30:95–99. 2012. View Article : Google Scholar : PubMed/NCBI
23	Tirino V, Desiderio V, Paino F, et al: Methods for cancer stem cell detection and isolation. Methods Mol Biol. 879:513–529. 2012. View Article : Google Scholar : PubMed/NCBI
24	Chinn SB, Darr OA, Peters RD, et al: The role of head and neck squamous cell carcinoma cancer stem cells in tumorigenesis, metastasis, and treatment failure. Front Endocrinol (Lausanne). 3:902012.PubMed/NCBI
25	Zhang SL, Wang YS, Zhou T, et al: Isolation and characterization of cancer stem cells from cervical cancer HeLa cells. Cytotechnology. 64:477–484. 2012. View Article : Google Scholar : PubMed/NCBI
26	Pham PV, Phan NL, Nguyen NT, et al: Differentiation of breast cancer stem cells by knockdown of CD44: promising differentiation therapy. J Transl Med. 9:2092011. View Article : Google Scholar : PubMed/NCBI
27	Zheng ZH, Yang Y, Lu XH, et al: Mycophenolic acid induces adipocyte-like differentiation and reversal of malignancy of breast cancer cells partly through PPARγ. Eur J Pharmacol. 658:1–8. 2011.PubMed/NCBI
28	Yan J, Luo D, Luo Y, et al: Induction of G1 arrest and differentiation in MDA-MB-231 breast cancer cell by boehmeriasin A, a novel compound from plant. Int J Gynecol Cancer. 16:165–170. 2006. View Article : Google Scholar : PubMed/NCBI
29	Chakraborty A, Bodipati N, Demonacos MK, et al: Long term induction by pterostilbene results in autophagy and cellular differentiation in MCF-7 cells via ROS dependent pathway. Mol Cell Endocrinol. 355:25–40. 2012. View Article : Google Scholar : PubMed/NCBI
30	Zhau HE, He H, Wang CY, et al: Human prostate cancer harbors the stem cell properties of bone marrow mesenchymal stem cells. Clin Cancer Res. 17:2159–2169. 2011. View Article : Google Scholar : PubMed/NCBI