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International Journal of Morphology

On-line version ISSN 0717-9502

Abstract

GHANBARI, Tara; AZADBAKHT, Mehri; VESI-RAYGANI, Asad  and  KHAZAEI, Mozafar. Isolation of Breast Cancer Stem Cell from MDA-MB231 Cell Line Using Vincristine. Int. J. Morphol. [online]. 2016, vol.34, n.4, pp.1197-1202. ISSN 0717-9502.  http://dx.doi.org/10.4067/S0717-95022016000400003.

Cancer has been considered as a stem cell disease. Suspension culture combined with anti-cancer drugs has recently been proposed for isolation of cancer stem cells (CSCs). In the current study, Vincristine as an anti-cancer drug combined with suspension culture was used for isolation and purification of CSCs from human breast cancer cell line (MDA-MB231). The cells were treated with different concentrations of vincristine (0, 2, 4, 6 and 8 ng/ml). Stem cells were identified with the expression of OCT4, nanog, SOX2 and nucleostemin genes by RT-PCR. Mammosphere forming unit was measured upon suspension culture containing EGF, bFGF, LIF, B27, insulin and BSA. The isolated mammospheres were investigated for CD44 expression. Results showed that 4 ng/ml of vincristine for 72 hours could be utilized as the best and most reliable dose which eliminates around 80 % of non-cancer stem cells with no destructive effect on CSCs' viability (P> 0.05). RT-PCR demonstrated that drug treated cells expressed OCT4, nanog, SOX2 and nucleostemin. Mammosphere formation unit of cells pretreated with vincristine was significantly higher than unpretreated ones (P>0.05). Immunofluorescence staining for CD44 depicted high expression of CSC marker among the isolated mammospheres. Vincristine combined with suspension culture can be considered as an appropriate method to isolate CSC.

Keywords : Cancer stem cell; Vincristine; Suspension culture; MDA-MB231.

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