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Characterization of breast cancer stem cells isolated from 4T1 cell line cultured in vitro

Do Thi Phuong, Nguyen Thi Nga, Nguyen Thi Cuc, Vu Thi Thu Phuong, Do Thi Thao

Abstract


Breast cancer is the most common cancer in women of which the main causes of death are its relapse and drug resistance. Recently, researchers clarified a small population of cancer cells holding the abilities of self-renewal, unlimited propagation responsible for tumor progression, metastasis, therapy resistance, and tumor recurrence. These cells are termed as cancer sterm cells (CSCs), possessing characteristics of stem cells. Thus, cancer stem cells have become to be a potential target for cancerous drug discovery. They can be used in a new research model for establishing novel therapy as well as sreening new compounds that target CSCs. In our studies, we isolated cancer stem cells from 4T1 cell line using serum-free medium and non-adherent culture surface. Those CSCs clones were formed after 10 culturing days from 1 initial cell. The clones could initiate subtumorsphere better than 4T1 cells themselves. There were 3 selected clones named as Đ3G5, Đ3C3 and Đ4E7 which had mammosphere formation efficiency over 30% (31.67%, 36.33%, and 35.33%, respectively). These clones showed the significant withstanding against ellipticine and/or doxorobicin stronger than that of  4T1 cells (P<0.05). The IC50 values (inhibition of cell growth at 50%) of elliticine was 0.89 ± 0.07 μg/mL and of doxorobicin was 0.58 ± 0.04 μg/mL toward 4T1 cell growth while those values were 2 - 3 folds higher in our isolated cancer stem cell clones (P<0.05). Those clones also exhibited strong expression of several typical genes of stem cells such as nanog and Sox2.


Keywords


4T1, cancer stem cell, cell clone, mammosphere, nanog, Sox2, tumorsphere

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References


Aparicio LA, Espinosa JC, Campelo RG, Veiga FG, Prado SD, Gallego GA (2007) Prostate carcinoma and stem cells. Clin Transl Oncol 9(2): 66–76.

Calvet CY, André FM, Mir LM (2014) The culture of cancer cell lines as tumorspheres does not systematically result in cancer stem cell enrichment. PLoS One 9(2): e89644.

Cao L, Zhou Y, Zhai B, Liao J, Xu W, Zhang R, Li J, Zhang Y, Chen L, Qian H, Wu M (2011) Sphere-forming cell subpopulations with cancer stem cell properties in human hepatoma cell lines. BMC Gastroenterol 11(1): 71.

Ceder JA, Jansson L, Ehrnström RA, Rönnstrand L, Abrahamsson PA (2008) The characterization of epithelial and stromal subsets of candidate stem/progenitor cells in the human adult prostate. Eur Urol 53(3): 524–31

Clevers H (2011) The cancer stem cell: premises, promises and challenges. Nat Med 7: 313–319.

Colleoni M, Viale G, Zahrieh D, Pruneri G, Gentilini O, Veronesi P, Gelber RD, Curigiano G, Torrisi R, Luini A, Intra M, Galimberti V, Renne G, Nolè F, Peruzzotti G, Goldhirsch A (2004) Chemotherapy is more effective in patients with breast cancer not expressing steroid hormone receptors: a study of preoperative treatment. Clin Cancer Res 10(19): 6622–6628.

Dontu G, Abdallah WM, Foley JM, Jackson KW, Clarke MF, Kawamura MJ, Wicha MS (2003) In vitro propagation and transcriptional profiling of human mammary stem/progenitor cells. Genes Dev 17(10): 1253–1270.

Eramo A, Lotti F, Sette G, Pilozzi E, Biffoni M, Di Virgilio A, Conticello C, Ruco L, Peschle C, De Maria R (2008) Identification and expansion of the tumorigenic lung cancer stem cell population. Cell Death Differ 15: 504–514.

Fillmore CM, Kuperwasser C (2008) Human breast cancer cell lines contain stem-like cells that self-renew, give rise to phenotypically diverse progeny and survive chemotherapy. Breast Cancer Res 10(2): R25. doi:10.1186/bcr1982.

Glinsky GV (2007) Stem cell origin of death-from-cancer phenotypes of human prostate and breast cancers. Stem Cell Rev 3(1): 79–93.

Ho MM, Ng AV, Lam S, Hung JY (2007) Side population in human lung cancer cell lines and tumors is enriched with stem-like cancer cells. Cancer Res 67(10): 4827–4833.

Li C, Heidt DG, Dalerba P, Burant CF, Zhang L, Adsay V, Wicha M, Clarke MF, Simeone DM (2007) Identification of pancreatic cancer stem cells. Cancer Res 67: 1030-1037. Ponti D, Costa A, Zaffaroni N, Pratesi G, Petrangolini G, Coradini D, Pilotti S, Pierotti MA, Daidone MG (2005) Isolation and in vitro propagation of tumorigenic breast cancer cells with stem/progenitor cell properties. Cancer Res 65(13): 5506–5511.

Prince ME, Sivanandan R, Kaczorowski A, Wolf GT, Kaplan MJ, Dalerba P, Weissman IL, Clarke MF, Ailles LE (2007) Identification of a subpopulation of cells with cancer stem cell properties in head and neck squamous cell carcinoma. Proc Natl Acad Sci 104: 973–978.

Rao GH, Liu HM, Li BW, Hao JJ, Yang YL, Wang MR, Wang XH, Wang J, Jin HJ, Du L, Chen Q (2013) Establishment of a human colorectal cancer cell line P6C with stem cell properties and resistance to chemotherapeutic drugs. Acta Pharmacol Sin 34(6): 793–804.

Vinogradov S, Wei X (2012) Cancer stem cells and drug resistance: the potential of nanomedicine. Nanomedicine (Lond) 7(4): 597–615.

Li Y, Kong D, Ahmad A, Bao B, Sarkar FH (2012) Pancreatic cancer stem cells: Emerging target for designing novel therapy. Cancer Lett

Yu C, Yao Z, Jiang Y, Keller ET (2012a) Prostate cancer stem cell biology. Minerva Urol Nefrol: 64(1):19–33.

Yu Y, Ramena G, Elble RC (2012b) The role of cancer stem cells in relapse of solid tumors. Front Biosci (Elite Ed) 4:1528–1541

Shi-cang Yu, Yi-fang Ping, Liang Yi, Zhi-hua Zhou, Jian-hong Chen, Xiao-hong Yao, Lei Gao, Ji Ming Wang, Xiu-wu Bian (2008) Isolation and characterization of cancer stem cells from a human glioblastoma cell line U87. Cancer Lett 265: 124–134.




DOI: https://doi.org/10.15625/1811-4989/16/3/10643 Display counter: Abstract : 269 views. PDF : 2 views.