Sorting of side population cells from multiple myeloma cell lines and analysis of their biological characteristics.
10.7534/j.issn.1009-2137.2014.03.032
- Author:
Xiao-Li ZHANG
1
;
Li-Na ZHANG
1
;
Hong-Ming HUANG
2
;
Run-Sheng DING
1
;
Wei SHI
3
;
Rui-Rong XU
1
;
Xiao-Tang YU
1
;
Sheng-Hua JIANG
1
Author Information
1. Department of Hematology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
2. Department of Hematology, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China. E-mail: hhmmmc@163.com.
3. Comprehensive Surgical Laboratory, Affiliated Hospital of Nantong University, Nantong 226001, Jiangsu Province, China.
- Publication Type:Journal Article
- MeSH:
Cell Line, Tumor;
Cytological Techniques;
methods;
Humans;
Multiple Myeloma;
Neoplastic Stem Cells;
cytology;
Side-Population Cells;
cytology
- From:
Journal of Experimental Hematology
2014;22(3):747-752
- CountryChina
- Language:Chinese
-
Abstract:
This study was aimed to sort the side population (SP) cells from human multiple myeloma cell lines, then detect the biological characteristics of those SP cells. After Hoechst33342 staining, intracellular Hoechst33342 fluorescence staining differences of myeloma cell lines observed by the fluorescence microscopy. The fluorescence-activated cell sorting (FACS) technology was used to isolate SP cells and main population (MP) cells; proliferative capacity in vitro was determined by cell growth curve; the cell colony forming ability was compared by colony forming test. The CD138 expression was detected by flow cytometry. The expression of ABCG2 mRNA was detected by reverse transcription PCR; CCK-8 assay and colony forming test were used to evaluate the effect of bortezomib on the cell proliferation, vitality and colony forming ability of the two populations. The results showed that the myeloma cell lines had a small proportion of SP cells, especially, RPMI 8226 cells accounted for the highest proportion of SP cells (7.10 ± 2.69)%, which have also been confirmed under the fluorescence microscope; the proliferative activity and cell colony forming ability of SP cells were significantly higher than those of MP cells (P < 0.05). The expression levels of CD138 in SP and MP cells were not significantly different (P > 0.05). RT-PCR results showed that SP cells expressed the drug-resistance gene ABCG2, but MP cells hardly express these genes. The inhibition rate of bortezomib on SP cells was significantly lower than that on MP cells (P < 0.05), however, the difference was not significant (P > 0.05) at bortezomib 40 nmol/L. Bortezomib could reduce colony formation in the both two cell populations, but more severe reduction appeared in the MP cells. It is concluded that the myeloma cell line contain a small amount of SP cells with the cancer stem cell characteristics.
