Abstract
The objective of this study is to investigate the chemoresistance of CD133+ cancer stem cells in Hep-2 cells of laryngeal cancer and detect the expression mRNA and protein levels of BMI-1 in CD133+ cells and CD133− cells. The response of Hep-2 cells to different chemotherapeutic agents was investigated, and the expression of CD133 was studied. Fluorescence-activated cell sorting analysis was used to identify CD133, and the CD133+ subset of cells was separated and analyzed chemotherapy resistance. Colony formation assays were studied and cells were injected subcutaneously into axillary fossa of node mice to measure the tumor-forming ability. RT-PCR and Western blot analyses were used to detect the expression levels of BMI-1 in the different subpopulation cells. It was concluded that chemotherapy enriched the CD133+ subpopulation 2-fourfold, relative to the untreated cells. 1.55 ± 0.28 % of Hep-2 cells were observed to be CD133+ cells. Flow cytometric analysis revealed that after the treatment with these chemotherapeutic agents, the expression of CD133 was up to 5.16 ± 0.86 %, 4.94 ± 0.58 %, 3.66 ± 0.59 %. After 5-FU treatment, the expression of CD133 was 6.7 ± 1.6 % relative to the untreated mice 2.6 ± 0.96 % by nude mice tumor xenograft model. CD133+ cancer stem cells were more resistant to chemotherapy; the proliferation capability and tumor-forming ability were no difference after chemotherapy. Semi-quantitative RT-PCR and Western blot analyses provided strong evidence that BMI-1 expression in CD133+ cells is different from CD133− cells remarkably. Taken together, it was confirmed that CD133+ cancer stem cells were chemoresistant and BMI-1 was highly expressed in these CD133+ cells.
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Acknowledgments
This work is supported by the National Natural Science Foundation of China (No.30973288), the Natural Science Foundation of Jilin Province of China (No.20130101151JC), and Doctoral Fund of Ministry of Education of China (No. 20120061120092).
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Dan Yu and Yan Liu have contributed equally to this work.
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Yu, D., Liu, Y., Yang, J. et al. Clinical Implications of BMI-1 in Cancer Stem Cells of Laryngeal Carcinoma. Cell Biochem Biophys 71, 261–269 (2015). https://doi.org/10.1007/s12013-014-0194-z
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DOI: https://doi.org/10.1007/s12013-014-0194-z