Separation and identification of tumor stem cells in human nasopharyngeal carcinoma cell lines and its mechanism of multidrug resistance(PDF)
《中国医学物理学杂志》[ISSN:1005-202X/CN:44-1351/R]
- Issue:
- 2018年第8期
- Page:
- 956-961
- Research Field:
- 医学生物物理
- Publishing date:
Info
- Title:
- Separation and identification of tumor stem cells in human nasopharyngeal carcinoma cell lines and its mechanism of multidrug resistance
- Author(s):
- LIU Chaoqun
- Department of Oncology, the Third Xiangya Hospital of Central South University, Changsha 410013, China
- Keywords:
- ; Keywords: nasopharyngeal carcinoma; malignant tumor; tumor stem cell; isolation and identification; multidrug resistance; immunocytochemistry; flow cytometry; nasopharyngeal carcinoma cells; in vitro differentiation; immunomagnetic cell separation technology
- PACS:
- R739.6
- DOI:
- DOI:10.3969/j.issn.1005-202X.2018.08.017
- Abstract:
- Abstract: Objective To study the method for separating and identifying tumor stem cells in human nasopharyngeal carcinoma cell lines, and investigate the mechanism of multidrug resistance. Methods The cell culture, subculture and identification of nasopharyngeal carcinoma cell lines (SUNE) were carried out, and the in vitro differentiation ability of CD133- cells and CD133+ cells in SUNE was detected with immunocytochemistry combined with flow cytometry. CD133+ tumor cells were purified with immunomagnetic cell separation technology. The in vitro proliferation ability of CD133+ cells was determined, and then compared with that of unsorted cells and CD133- cells. The drug resistance of CD133+ cells in the treatment of cisplatin and paclitaxel was determined by CCK-8 method. ten ICR mice aged 4 weeks were injected subcutaneously with human nasopharyngeal carcinoma cell lines and normal cell lines, and the mechanism of multidrug resistance was analyzed. Results The SUNE isolated and cultured in serum-containing medium were adherent growth, with active growth. After SUNE being cultured for 2-3 d, inverted microscope showed that the cells were spindle, flat, with good gloss. After SUNE being cultured for about 2 weeks, the cell fusion bottle was 80.00%; and with the use of flow cytometry, antigen CD133 was detected on about 0.35% of cell surface membrane. Immunocytochemistry showed that SUNE cells were adherent growth, some of which were combined with CD133- in SUNE, appearing orange red and globose under fluorescence microscope. The cells after immunomagnetic cell separation were added to the medium, and the cells were single cells, spherical, in suspended growth. With the prolonging of culture time, cells began to appear nodular growth, increase in volume and cell quantity. The cells were adherent growth after being cultured for 24 h, and began to grow in clusters after 7 d of culture. The number of unsorted cells, CD133- tumor cells and CD133+ tumor cells were increased with the extension of time, and the proliferation of CD133+ tumor cells was significantly higher than that of CD133- tumor cells and unsorted cells (P<0.05). The tumors were found in all 10 mice, and the volume of human nasopharyngeal carcinoma cell lines in the right side were significantly larger than that of the common cell lines (P<0.05). Conclusion CD133+ cells in nasopharyngeal carcinoma stem cells have a strong resistance to chemotherapeutic drugs.
Last Update: 2018-07-26