Characterization of side population cells isolated from the colon cancer cell line SW480

Xiong,Binghong; Ma,Li; Hu,Xiang; Zhang,Caiquan; Cheng,Yong

doi:10.3892/ijo.2014.2498

Characterization of side population cells isolated from the colon cancer cell line SW480

Authors:
- Binghong Xiong
- Li Ma
- Xiang Hu
- Caiquan Zhang
- Yong Cheng
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Affiliations: Department of General Surgery, The First Affiliated Hospital of Chongqing Medical University, Yuanjiagang, Yuzhong, Chongqing 400016, P.R. China , Department of Internal Medicine, Chongqing Huaxi Hospital, Banan, Chongqing 400054, P.R. China
Published online on: June 13, 2014 https://doi.org/10.3892/ijo.2014.2498
Pages: 1175-1183

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Abstract

Side population (SP) cells may play a crucial role in tumorigenesis and the recurrence of cancer. Many types of cell lines and tissues have demonstrated the presence of SP cells, including colon cancer cell lines. This study aimed to identify cancer stem cells (CSCs) in the SP of the colon cancer cell line SW480. SP cells were isolated by fluorescence-activated cell sorting (FACS), followed by serum-free medium (SFM) culture. The self-renewal, differentiated progeny, clone formation, proliferation, invasion ability, cell cycle, chemosensitivity and tumorigenic properties in SP and non-SP (NSP) cells were investigated through in vitro culture and in vivo serial transplantation. The expression profiles of ATP-binding cassette (ABC) protein transporters and stem cell‑related genes were examined by RT-PCR and western blot analysis. The human colon cancer cell lines SW480, Lovo and HCT116 contain 1.1±0.10, 0.93±0.11 and 1.33±0.05% SP cells, respectively. Flow cytometry analysis revealed that SP cells could differentiate into SP and NSP cells. SP cells had a higher proliferation potency and CFE than NSP cells. Compared to NSP cells, SP cells were also more resistant to CDDP and 5-FU, and were more invasive and displayed increased tumorigenic ability. Moreover, SP cells showed higher mRNA and protein expression of ABCG2, MDR1, OCT-4, NANOG, SOX-2, CD44 and CD133. SP cells isolated from human colon cancer cell lines harbor CSC properties that may be related to the invasive potential and therapeutic resistance of colon 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	Jemal A, Siegel R, Xu J, et al: Cancer statistics. CA Cancer J Clin. 60:277–300. 2010.
3	Zhang YL, Zhang ZS, Wu BP, et al: Early diagnosis for colorectal cancer in China. World J Gastroenterol. 8:21–25. 2002.
4	Parkin DM, Bray F, Ferlay J, et al: Global cancer statistics, 2002. CA Cancer J Clin. 55:74–108. 2005. View Article : Google Scholar
5	Clarke MF, Dick JE, Dirks PB, et al: Cancer stem cells: perspectives on current status and future directions - AACR Workshop on Cancer Stem Cells. Cancer Res. 66:9339–9344. 2006. View Article : Google Scholar
6	Mittal S, Mifflin R and Powell DW: Cancer stem cells: the other face of Janus. Am J Med Sc. 338:107–112. 2009. View Article : Google Scholar : PubMed/NCBI
7	Ricci-Vitiani L, Lombardi DG, Pilozzi E, et al: Identification and expansion of human colon-cancer-initiating cells. Nature. 445:111–115. 2007. View Article : Google Scholar : PubMed/NCBI
8	O’Brien CA, Pollett A, Gallinger S, et al: A human colon cancer cell capable of initiating tumour growth in immunodeficient mice. Nature. 445:106–110. 2007.PubMed/NCBI
9	Dalerba P, Dylla SJ, Park IK, et al: Phenotypic characterization of human colorectal cancer stem cells. Proc Natl Acad Sci USA. 104:10158–10163. 2007. View Article : Google Scholar : PubMed/NCBI
10	Goodell MA, Brose K, Paradis G, et al: Isolation and functional properties of murine hematopoietic stem cells that are replicating in vivo. J Exp Med. 183:1797–1806. 1996. View Article : Google Scholar : PubMed/NCBI
11	Kondo T, Setoguchi T and Taga T: Persistence of a small subpopulation of cancer stem-like cells in the C6 glioma cell line. Proc Natl Acad Sci USA. 101:781–786. 2004. View Article : Google Scholar : PubMed/NCBI
12	Hirschmann-Jax C, Foster AE, Wulf GG, et al: A distinct ‘side population’ of cells with high drug efflux capacity in human tumor cells. Proc Natl Acad Sci USA. 101:14228–14233. 2004.
13	Wang J, Guo LP, Chen LZ, et al: Identification of cancer stem cell-like side population cells in human nasopharyngeal carcinoma cell line. Cancer Res. 67:3716–3724. 2007. View Article : Google Scholar : PubMed/NCBI
14	Zhang P, Zhang Y, Mao L, et al: Side population in oral squamous cell carcinoma possesses tumor stem cell phenotypes. Cancer Lett. 277:227–234. 2009. View Article : Google Scholar : PubMed/NCBI
15	Yanamoto S, Kawasaki G, Yamada S, et al: Isolation and characterization of cancer stem-like side population cells in human oral cancer cells. Oral Oncol. 47:855–860. 2011. View Article : Google Scholar : PubMed/NCBI
16	Huang D, Gao Q, Guo L, et al: Isolation and identification of cancer stem-like cells in esophageal carcinoma cell lines. Stem Cells Dev. 18:465–473. 2009. View Article : Google Scholar : PubMed/NCBI
17	Haraguchi N, Utsunomiya T, Inoue H, et al: Characterization of a side population of cancer cells from human gastrointestinal system. Stem Cells. 24:506–513. 2006. View Article : Google Scholar : PubMed/NCBI
18	Li R, Wu X, Wei H, et al: Characterization of side population cells isolated from the gastric cancer cell line SGC-7901. Oncol Lett. 5:877–883. 2013.PubMed/NCBI
19	She JJ, Zhang PG, Wang X, et al: Side population cells isolated from KATO III human gastric cancer cell line have cancer stem cell-like characteristics. World J Gastroenterol. 18:4610–4617. 2012. View Article : Google Scholar : PubMed/NCBI
20	Fukuda K, Saikawa Y, Ohashi M, et al: Tumor initiating potential of side population cells in human gastric cancer. Int J Oncol. 34:1201–1207. 2009.PubMed/NCBI
21	Zhang H, Xi H, Cai A, et al: Not all side population cells contain cancer stem-like cells in human gastric cancer cell lines. Dig Dis Sci. 58:132–139. 2013. View Article : Google Scholar
22	Chiba T, Kita K, Zheng YW, Yokosuka O, Saisho H, Iwama A, Nakauchi H and Taniguchi H: Side population purified from hepatocellular carcinoma cells harbors cancer stem cell-like properties. Hepatology. 44:240–251. 2006. View Article : Google Scholar : PubMed/NCBI
23	Shi GM, Xu Y, Fan J, et al: Identification of side population cells in human hepatocellular carcinoma cell lines with stepwise metastatic potentials. J Cancer Res Clin Oncol. 134:1155–1163. 2008. View Article : Google Scholar : PubMed/NCBI
24	Li CW, Heidt DG, Dalerba P, et al: Identification of pancreatic cancer stem cells. Cancer Res. 67:1030–1037. 2007. View Article : Google Scholar : PubMed/NCBI
25	Zhou J, Wang CY, Liu T, et al: Persistence of side population cells with high drug efflux capacity in pancreatic cancer. World J Gastroenterol. 14:925–930. 2008. View Article : Google Scholar : PubMed/NCBI
26	Sung JM, Cho HJ, Yi H, et al: Characterization of a stem cell population in lung cancer A549 cells. Biochem Biophys Res Commun. 371:163–167. 2008. View Article : Google Scholar : PubMed/NCBI
27	Zhang AM, Fan Y, Yao Q, et al: Identification of a cancer stem-like population in the Lewis lung cancer cell line. Asian Pac J Cancer Prev. 13:761–766. 2012. View Article : Google Scholar : PubMed/NCBI
28	Ho MM, Ng AV, Lam S, et al: Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells. Cancer Res. 67:4827–4833. 2007. View Article : Google Scholar : PubMed/NCBI
29	Hiraga T, Ito S and Nakamura H: Side population in MDA-MB-231 human breast cancer cells exhibits cancer stem cell-like properties without higher bone-metastatic potential. Oncol Rep. 25:289–296. 2011.PubMed/NCBI
30	Nakanishi T, Chumsri S, Khakpour N, et al: Side-population cells in luminal-type breast cancer have tumour-initiating cell properties, and are regulated by HER2 expression and signaling. Br J Cancer. 102:815–826. 2010. View Article : Google Scholar : PubMed/NCBI
31	Szotek PP, Pieretti-Vanmarcke R, Masiakos PT, et al: Ovarian cancer side population defines cells with stem cell-like characteristics and Mullerian Inhibiting Substance responsiveness. Proc Natl Acad Sci USA. 103:11154–11159. 2006. View Article : Google Scholar : PubMed/NCBI
32	Patrawala L, Calhoun T, Schneider-Broussard R, et al: Side population is enriched in tumorigenic, stem-like cancer cells, whereas ABCG2⁺ and ABCG2⁻ cancer cells are similarly tumorigenic. Cancer Res. 65:6207–6219. 2005. View Article : Google Scholar : PubMed/NCBI
33	Chen Y, Zhao J, Luo Y, Wang Y, et al: Isolation and identification of cancer stem-like cells from side population of human prostate cancer cells. J Huazhong Univ Sci Technolog Med Sci. 32:697–703. 2012. View Article : Google Scholar : PubMed/NCBI
34	Burkert J, Otto WR and Wright NA: Side populations of gastrointestinal cancers are not enriched in stem cells. J Pathol. 214:564–573. 2008. View Article : Google Scholar : PubMed/NCBI
35	Xu XT, Xu Q, Tong JL, et al: MicroRNA expression profiling identifies miR-328 regulates cancer stem cell-like SP cells in colorectal cancer. Br J Cancer 2012. 106:1320–1330. 2012.PubMed/NCBI
36	Triel C, Vestergaard ME, Bolund L, et al: Side population cells in human and mouse epidermis lack stem cell characteristics. Exp Cell Res. 295:79–90. 2004. View Article : Google Scholar : PubMed/NCBI
37	Morita Y, Ema H, Yamazaki S, et al: Non-side-population hematopoietic stem cells in mouse bone marrow. Blood. 108:2850–2856. 2006. View Article : Google Scholar : PubMed/NCBI
38	Dean M, Fojo T and Bates S: Tumour stem cells and drug resistance. Nat Rev Cancer. 5:275–284. 2005. View Article : Google Scholar
39	Behbod F, Xian W, Shaw CA, et al: Transcriptional profiling of mammary gland side population cells. Stem Cells. 24:1065–1074. 2006. View Article : Google Scholar : PubMed/NCBI
40	De Paiva CS, Chen Z, Corrales RM, et al: ABCG2 transporter identifies a population of clonogenic human limbal epithelial cells. Stem Cells. 23:63–73. 2005.PubMed/NCBI
41	Woodward WA, Chen MS, Behbod F, et al: On mammary stem cells. J Cell Sci. 118:3585–3594. 2005. View Article : Google Scholar : PubMed/NCBI
42	Marx J: Cancer research. Mutant stem cells may seed cancer. Science. 301:1308–1310. 2003. View Article : Google Scholar : PubMed/NCBI
43	Miyake K, Mickley L, Litman T, et al: Molecular cloning of cDNAs which are highly overexpressed in mitoxantrone-resistant cells: demonstration of homology to ABC transport genes. Cancer Res. 59:8–13. 1999.
44	Kim M, Turnquist H, Jackson J, et al: The multidrug resistance transporter ABCG2 breast cancer resistance protein 1 effluxes Hoechst 33342 and is overexpressed in hematopoietic stem cells. Clin Cancer Res. 8:22–28. 2002.PubMed/NCBI
45	Chaudhary PM and Roninson IB: Expression and activity of P-glycoprotein, a multidrug efflux pump, in human hematopoietic stem cells. Cell. 66:85–94. 1991. View Article : Google Scholar : PubMed/NCBI
46	Kim J, Chu J, Shen X, et al: An extended transcriptional network for pluripotency of embryonic stem cells. Cell. 132:1049–1061. 2008. View Article : Google Scholar : PubMed/NCBI
47	Naor D, Nedvetzki S, Golan I, et al: CD44 in cancer. Crit Rev Clin Lab Sci. 39:527–579. 2002. View Article : Google Scholar
48	Ponta H, Sherman L and Herrlich PA: CD44: from adhesion molecules to signalling regulators. Nat Rev Mol Cell Biol. 4:33–45. 2003. View Article : Google Scholar : PubMed/NCBI
49	Takaishi S, Okumura T, Tu S, et al: Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells. 27:1006–1020. 2009. View Article : Google Scholar : PubMed/NCBI
50	Yin AH, Miraglia S, Zanjani ED, et al: AC133, a novel marker for human hematopoietic stem and progenitor cells. Blood. 90:5002–5012. 1997.PubMed/NCBI
51	Shmelkov SV, Butler JM, Hooper AT, et al: CD133 expression is not restricted to stem cells, and both CD133⁺ and CD133⁻ metastatic colon cancer cells initiate tumors. J Clin Invest. 118:2111–2120. 2008.PubMed/NCBI
52	Meng X, Li M, Wang X, et al: Both CD133⁺ and CD133⁻ subpopulations of A549 and H446 cells contain cancer-initiating cells. Cancer Sci. 100:1040–1046. 2009.