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Expression of stem cell markers in the side population of ovarian cancer SKOV3 cells
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Abstract:
Objective: To study the differential expression of ovarian cancer stem cell specific markers in the side population (SP) and non-SP of human epithelial ovarian cancer SKOV3 cells. Methods: The SP of SKOV3 cells was identified and sorted by flow cytometry using the blue fluorescent dye Hoechst 33342. The expression of CD133, CD117, CD44, ABCG2 and ALDH2 at the protein level in SP and non-SP cells was assessed by flow cytometry. The expression of ALDH1, ABCG2, NANOG, OCT4, SOX2, CD133 and CD117 at the mRNA level in SP and non-SP cells was assessed by RT-PCR. Results: In SKOV3 cells, SP accounted for (1.56±0.35)%. ALDH1 and ABCG2 proteins were detected in (87.3±5.76)% and (29.48±4.43)% of SP cells respectively, both significantly higher than those in non-SP cells ( P <0.05). CD44 protein was detected in greater than 99% of cells in both subpopulations ( P >0.05). Neither CD133 nor CD117 protein was detected in both subpopulations. The mRNA levels of ALDH1, ABCG2, NANOG, OCT4 and SOX2 in SP cells were increased by 21.03 ( P =0.001), 3.14( P =0.001), 23.94 ( P =0.001), 10.73 ( P =0.009), 21.46 ( P =0.001) folds , respectively, as compared with the levels in Non-SP cells. CD133 and CD117 transcripts were detected in neither sub-population of SKOV 3 cells. Conclusion: There is a SP in human ovarian cancer SKOV3 cells expressing stem cell markers ALDH1 and ABCG2 at the protein level and NANOG, OCT4 , and SOX2 at the mRNA level like ovarian cancer stem cells. This SP of SKOV3 cells may offer a very useful tool for ovarian cancer stem cell research.
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Project Supported:
Project supported by Key Projects of Guangxi Health Bureau（No.2010074）, the Natural Science Foundation of Guangxi Province（No .2012GXNSFAA053134）, and the Innovation Project of Guangxi Postgraduate Education Fund（No. YCSZ2012045）
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