Isolation and characterization of side population cells from the human ovarian cancer cell line SK-OV-3

Ruan,Zhengyi; Liu,Jianhua; Kuang,Yanping

doi:10.3892/etm.2015.2836

Isolation and characterization of side population cells from the human ovarian cancer cell line SK-OV-3

Authors:
- Zhengyi Ruan
- Jianhua Liu
- Yanping Kuang
View Affiliations

Affiliations: Department of Gynecology, Shanghai Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China, Department of Assisted Reproduction, Shanghai Ninth People's Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200011, P.R. China
Published online on: October 30, 2015 https://doi.org/10.3892/etm.2015.2836
Pages: 2071-2078
Copyright: © Ruan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Ovarian cancer (OC) is the most malignant type of gynecological tumor due to its high recurrence rate following initial treatment. Previous studies have indicated that cancer stem cells (CSCs) may be a potential cause underlying the high proportion of recurrence. Side population (SP) cells isolated from cancer cell lines have been shown to exhibit characteristics associated with CSCs, but studies on SP cells in human ovarian SK‑OV‑3 cell line are limited. In the present study, the SP cell fraction (4.83% of the total cell population) was isolated using flow cytometry, and analyzed by immunocytochemical analysis and reverse transcription‑quantitative polymerase chain reaction. The results showed that SP cells exhibited a high mean fluorescence intensity for CD44, a CSC marker, in addition to elevated expression of the CSCs‑associated genes, ATP‑binding cassette sub‑family G member 2 and Nestin. These findings indicated the stem cell‑like features of the SP cells. Furthermore, a colony formation test showed that the isolated SP cells possessed a marked capacity for self‑regeneration and proliferation. In addition, a cell cycle assay involving cisplatin indicated that the SP cells were strongly resistant to chemotherapy. In conclusion, the present results suggested that SP cells isolated from the SK‑OV‑3 cell line exhibited properties typically associated with CSCs. Therefore, the isolated SP cells may be used to provide novel insight into potential therapies against OC.

View Figures

View References

1	Ruscito I, Dimitrova D, Vasconcelos I, Gellhaus K, Schwachula T, Bellati F, Zeillinger R, Benedetti-Panici P, Vergote I, Mahner S, et al: BRCA1 gene promoter methylation status in high-grade serous ovarian cancer patients-a study of the tumour bank ovarian cancer (TOC) and ovarian cancer diagnosis consortium (OVCAD). Eur J Cancer. 50:2090–2098. 2014. View Article : Google Scholar : PubMed/NCBI
2	Chen J, Wang J, Zhang Y, Chen D, Yang C, Kai C, Wang X, Shi F and Dou J: Observation of ovarian cancer stem cell behavior and investigation of potential mechanisms of drug resistance in three-dimensional cell culture. J Biosci Bioeng. 118:214–222. 2014. View Article : Google Scholar : PubMed/NCBI
3	Cannistra SA: Cancer of the ovary. N Engl J Med. 351:2519–2529. 2004. View Article : Google Scholar : PubMed/NCBI
4	Purdie DM, Bain CJ, Siskind V, Webb PM and Green AC: Ovulation and risk of epithelial ovarian cancer. Int J Cancer. 104:228–232. 2003. View Article : Google Scholar : PubMed/NCBI
5	Boyd J and Rubin SC: Hereditary ovarian cancer: Molecular genetics and clinical implications. Gynecol Oncol. 64:196–206. 1997. View Article : Google Scholar : PubMed/NCBI
6	Goff BA, Mandel L, Muntz HG and Melancon CH: Ovarian carcinoma diagnosis. Cancer. 89:2068–2075. 2000. View Article : Google Scholar : PubMed/NCBI
7	Meunier L, Puiffe ML, Le Page C, Lali-Mouhim A, Chevrette M, Tonin PN, Provencher DM and Mes-Masson AM: Effect of ovarian cancer ascites on cell migration and gene expression in an epithelial ovarian cancer in vitro model. Transl Oncol. 3:230–238. 2010. View Article : Google Scholar : PubMed/NCBI
8	Fishman DA, Cohen L, Blank SV, Shulman L, Singh D, Bozorgi K, Tamura R, Timor-Tritsch I and Schwartz PE: The role of ultrasound evaluation in the detection of early-stage epithelial ovarian cancer. Am J Obstet Gynecol. 192:1214–1221; discussion 1221–1222. 2005. View Article : Google Scholar : PubMed/NCBI
9	Chobanian N and Dietrich CS III: Ovarian cancer. Surg Clin North Am. 88:285–299. 2008. View Article : Google Scholar : PubMed/NCBI
10	Naora H and Montell DJ: Ovarian cancer metastasis: Integrating insights from disparate model organisms. Nat Rev Cancer. 5:355–366. 2005. View Article : Google Scholar : PubMed/NCBI
11	Eyler CE and Rich JN: Survival of the fittest: Cancer stem cells in therapeutic resistance and angiogenesis. J Clin Oncol. 26:2839–2845. 2008. View Article : Google Scholar : PubMed/NCBI
12	Klonisch T, Wiechec E, Hombach-Klonisch S, Ande SR, Wesselborg S, Schulze-Osthoff K and Los M: Cancer stem cell markers in common cancers-therapeutic implications. Trends Mol Med. 14:450–460. 2008. View Article : Google Scholar : PubMed/NCBI
13	Bonnet D and Dick JE: Human acute myeloid leukemia is organized as a hierarchy that originates from a primitive hematopoietic cell. Nat Med. 3:730–737. 1997. View Article : Google Scholar : PubMed/NCBI
14	Dexter DL, Spremulli EN, Fligiel Z, Barbosa JA, Vogel R, VanVoorhees A and Calabresi P: Heterogeneity of cancer cells from a single human colon carcinoma. Am J Med. 71:949–956. 1981. View Article : Google Scholar : PubMed/NCBI
15	Al-Hajj M and Clarke MF: Self-renewal and solid tumor stem cells. Oncogene. 23:7274–7282. 2004. View Article : Google Scholar : PubMed/NCBI
16	Bonner WA, Hulett HR, Sweet RG and Herzenberg LA: Fluorescence activated cell sorting. Rev Sci Instrum. 43:404–409. 1972. View Article : Google Scholar : PubMed/NCBI
17	Shi GM, Xu Y, Fan J, Zhou J, Yang XR, Qiu SJ, Liao Y, Wu WZ, Ji Y, Ke AW, 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
18	Ojha R, Jha V, Singh SK and Bhattacharyya S: Autophagy inhibition suppresses the tumorigenic potential of cancer stem cell enriched side population in bladder cancer. Biochim Biophys Acta. 1842:2073–2086. 2014. View Article : Google Scholar : PubMed/NCBI
19	Yanamoto S, Kawasaki G, Yamada S, Yoshitomi I, Kawano T, Yonezawa H, Rokutanda S, Naruse T and Umeda M: 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
20	Kobayashi Y, Seino K, Hosonuma S, Ohara T, Itamochi H, Isonishi S, Kita T, Wada H, Kojo S and Kiguchi K: Side population is increased in paclitaxel-resistant ovarian cancer cell lines regardless of resistance to cisplatin. Gynecol Oncol. 121:390–394. 2011. View Article : Google Scholar : PubMed/NCBI
21	Chung WM, Chang WC, Chen L, Lin TY, Chen LC, Hung YC and Ma WL: Ligand-independent androgen receptors promote ovarian teratocarcinoma cell growth by stimulating self-renewal of cancer stem/progenitor cells. Stem Cell Res. 13:24–35. 2014. View Article : Google Scholar : PubMed/NCBI
22	Shah MM and Landen CN: Ovarian cancer stem cells: Are they real and why are they important? Gynecol Oncol. 132:483–489. 2014. View Article : Google Scholar : PubMed/NCBI
23	Provencher DM, Lounis H, Champoux L, Tétrault M, Manderson EN, Wang JC, Eydoux P, Savoie R, Tonin PN and Mes-Masson AM: Characterization of four novel epithelial ovarian cancer cell lines. Vitro Cell Dev Biol Anim. 36:357–361. 2000. View Article : Google Scholar
24	Kaur M, Ita KB, Popova IE, Parikh SJ and Bair DA: Microneedle-assisted delivery of verapamil hydrochloride and amlodipine besylate. Eur J Pharm Biopharm. 86:284–291. 2014. View Article : Google Scholar : PubMed/NCBI
25	Lecoeur H: Nuclear apoptosis detection by flow cytometry: Influence of endogenous endonucleases. Exp Cell Res. 277:1–14. 2002. View Article : Google Scholar : PubMed/NCBI
26	Prochazka L, Tesarik R and Turanek J: Regulation of alternative splicing of CD44 in cancer. Cell Signal. 26:2234–2239. 2014. View Article : Google Scholar : PubMed/NCBI
27	Cao Q, Zhang J, Liu H, Wu Q, Chen J and Chen GQ: The mechanism of anti-osteoporosis effects of 3-hydroxybutyrate and derivatives under simulated microgravity. Biomaterials. 35:8273–8283. 2014. View Article : Google Scholar : PubMed/NCBI
28	Manoharan-Valerio M, Ortiz C, Quiñones J, Feliciano G, Diaz A, Rivera D and Castro M: Determination of the IC₅₀ and LD₅₀ of calcium sulfide (CaS) clusters on malignant carcinoma and normal fibroblasts cell lines (LB580). The FASEB Journal. 28:LB5802014.
29	Poff AM, Ari C, Arnold P, Seyfried TN and D'Agostino DP: Ketone supplementation decreases tumor cell viability and prolongs survival of mice with metastatic cancer. Int J Cancer. 135:1711–1720. 2014. View Article : Google Scholar : PubMed/NCBI
30	Eissing N, Heger L, Baranska A, Cesnjevar R, Büttner-Herold M, Söder S, Hartmann A, Heidkamp GF and Dudziak D: Easy performance of 6-color confocal immunofluorescence with 4-laser line microscopes. Immunol Lett. 161:1–5. 2014. View Article : Google Scholar : PubMed/NCBI
31	Barber HR: Ovarian cancer. CA Cancer J Clin. 36:149–184. 1986. View Article : Google Scholar : PubMed/NCBI
32	Mueller H and Hahn M: Cytoreductive surgery plus intraperitoneal hyperthermic perfusion in the treatment of recurrent epithelial ovarian cancer: Oral Presentation 00012. Int J Gynecol Cancer. 15:542005.
33	Koch U, Krause M and Baumann M: Cancer stem cells at the crossroads of current cancer therapy failures-radiation oncology perspective. Semin Cancer Biol. 20:116–124. 2010. View Article : Google Scholar : PubMed/NCBI
34	Shigdar S, Lin J, Li Y, Yang CJ, Wei M, Zhus Y, Liu H and Duan W: Cancer stem cell targeting: The next generation of cancer therapy and molecular imaging. Ther Deliv. 3:227–244. 2012. View Article : Google Scholar : PubMed/NCBI
35	Espinoza LA, Albanese C and Rodriguez OC: The potential target therapy of prostate cancer stem cells. Prostate Cancer - From Bench to Bedside. Spiess PE: InTech. 2011.
36	Scatena R, Bottoni P, Pontoglio A and Giardina B: Cancer stem cells: The development of new cancer therapeutics. Expert Opin Biol Ther. 11:875–892. 2011. View Article : Google Scholar : PubMed/NCBI
37	Visvader JE and Lindeman GJ: Cancer stem cells in solid tumours: Accumulating evidence and unresolved questions. Nat Rev Cancer. 8:755–768. 2008. View Article : Google Scholar : PubMed/NCBI
38	Clevers H: The cancer stem cell: Premises, promises and challenges. Nat Med. 17:313–319. 2011. View Article : Google Scholar : PubMed/NCBI
39	Li XX, Wang J, Wang HL, Wang W, Yin XB, Li QW, Chen YY and Yi J: Characterization of cancer stem-like cells derived from a side population of a human gallbladder carcinoma cell line, SGC-996. Biochem Biophys Res Commun. 419:728–734. 2012. View Article : Google Scholar : PubMed/NCBI
40	Szotek PP, Pieretti-Vanmarcke R, Masiakos PT, Dinulescu DM, Connolly D, Foster R, Dombkowski D, Preffer F, Maclaughlin DT and Donahoe PK: 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
41	Hu L, McArthur C and Jaffe RB: Ovarian cancer stem-like side-population cells are tumourigenic and chemoresistant. Br J Cancer. 102:1276–1283. 2010. View Article : Google Scholar : PubMed/NCBI
42	Spring FA, Dalchau R, Daniels GL, Mallinson G, Judson PA, Parsons SF, Fabre JW and Anstee DJ: The Ina and Inb blood group antigens are located on a glycoprotein of 80,000 MW (the CDw44 glycoprotein) whose expression is influenced by the In(Lu) gene. Immunology. 64:37–43. 1988.PubMed/NCBI
43	Ricardo S, Vieira AF, Gerhard R, Leitão D, Pinto R, Cameselle-Teijeiro JF, Milanezi F, Schmitt F and Paredes J: Breast cancer stem cell markers CD44, CD24 and ALDH1: Expression distribution within intrinsic molecular subtype. J Clin Pathol. 64:937–946. 2011. View Article : Google Scholar : PubMed/NCBI
44	Lee CJ, Dosch J and Simeone DM: Pancreatic cancer stem cells. J Clin Oncol. 26:2806–2812. 2008. View Article : Google Scholar : PubMed/NCBI
45	Du L, Wang H, He L, Zhang J, Ni B, Wang X, Jin H, Cahuzac N, Mehrpour M, Lu Y and Chen Q: CD44 is of functional importance for colorectal cancer stem cells. Clin Cancer Res. 14:6751–6760. 2008. View Article : Google Scholar : PubMed/NCBI
46	Takaishi S, Okumura T, Tu S, Wang SS, Shibata W, Vigneshwaran R, Gordon SA, Shimada Y and Wang TC: Identification of gastric cancer stem cells using the cell surface marker CD44. Stem Cells. 27:1006–1020. 2009. View Article : Google Scholar : PubMed/NCBI
47	Weng D, Song B, Durfee J, Sugiyama V, Wu Z, Koido S, Calderwood SK and Gong J: Induction of cytotoxic T lymphocytes against ovarian cancer-initiating cells. Int J Cancer. 129:1990–2001. 2011. View Article : Google Scholar : PubMed/NCBI
48	Au A, Aziz Baba A, Goh AS, Wahid Fadilah SA, Teh A, Rosline H and Ankathil R: Association of genotypes and haplotypes of multi-drug transporter genes ABCB1 and ABCG2 with clinical response to imatinib mesylate in chronic myeloid leukemia patients. Biomed Pharmacother. 68:343–349. 2014. View Article : Google Scholar : PubMed/NCBI
49	Sodani K, Patel A, Anreddy N, Singh S, Yang DH, Kathawala RJ, Kumar P, Talele TT and Chen ZS: Telatinib reverses chemotherapeutic multidrug resistance mediated by ABCG2 efflux transporter in vitro and in vivo. Biochem Pharmacol. 89:52–61. 2014. View Article : Google Scholar : PubMed/NCBI
50	Castillo V, Valenzuela R, Huidobro C, Contreras HR and Castellon EA: Functional characteristics of cancer stem cells and their role in drug resistance of prostate cancer. Int J Oncol. 45:985–994. 2014.PubMed/NCBI
51	Yanamoto S, Yamada S, Takahashi H, Naruse T, Matsushita Y, Ikeda H, Shiraishi T, Seki S, Fujita S, Ikeda T, et al: Expression of the cancer stem cell markers CD44v6 and ABCG2 in tongue cancer: Effect of neoadjuvant chemotherapy on local recurrence. Int J Oncol. 44:1153–1162. 2014.PubMed/NCBI
52	El Deeb NM and Abdelzaher E: Stem cell markers OCT4 and nestin in laryngeal squamous cell carcinoma and their relation to survivin expression. Pathol Res Pract. 210:751–758. 2014. View Article : Google Scholar : PubMed/NCBI
53	Krüger K, Stefansson IM, Collett K, Arnes JB, Aas T and Akslen LA: Microvessel proliferation by co-expression of endothelial nestin and Ki-67 is associated with a basal-like phenotype and aggressive features in breast cancer. Breast. 22:282–288. 2013. View Article : Google Scholar : PubMed/NCBI
54	Ishiwata T, Matsuda Y and Naito Z: Nestin in gastrointestinal and other cancers: Effects on cells and tumor angiogenesis. World J Gastroenterol. 17:409–418. 2011. View Article : Google Scholar : PubMed/NCBI
55	Wan G, Zhou L, Xie M, Chen H and Tian J: Characterization of side population cells from laryngeal cancer cell lines. Head Neck. 32:1302–1309. 2010. View Article : Google Scholar : PubMed/NCBI
56	Dou J, Wen P, Hu W, Li Y, Wu Y, Liu C, Zhao F, Hu K, Wang J, Jiang C, et al: Identifying tumor stem-like cells in mouse melanoma cell lines by analyzing the characteristics of side population cells. Cell Biol Int. 33:807–815. 2009. View Article : Google Scholar : PubMed/NCBI
57	Zhang JL, Wang Z, Hu W, Chen SS, Lou XE and Zhou HJ: DHA regulates angiogenesis and improves the efficiency of CDDP for the treatment of lung carcinoma. Microvasc Res. 87:14–24. 2013. View Article : Google Scholar : PubMed/NCBI
58	Fan LM and Li JM: Evaluation of methods of detecting cell reactive oxygen species production for drug screening and cell cycle studies. J Pharmacol Toxicol Methods. 70:40–47. 2014. View Article : Google Scholar : PubMed/NCBI
59	Matsumoto A and Nakayama KI: Role of key regulators of the cell cycle in maintenance of hematopoietic stem cells. Biochim Biophys Acta. 1830:2335–2344. 2013. View Article : Google Scholar : PubMed/NCBI