Influences of co-culture with primary bone marrow stromal cells on imatinib sensitivity and cell cycles of K562 cells

doi:10.3969/j.issn.2095-4344.2014.28.004

Abstract

Abstract:

BACKGROUND: Leukemia cells can obtain drug resistance phenotype mediated by adhesion to bone marrow stromal cells. But, for chronic myelogenous leukemia with adhesion functional defects, the role and mechanism of bone marrow stromal cells in imatinib-resistant formation remain unclear.
OBJECTIVE: To construct the co-cultured model of bone marrow stromal cells–K562 cells and to investigate the influences of the co-culture with bone marrow stromal cells from the patients with chronic myelogenous leukemia on imatinib sensitivity of K562 cells and cell cycle.
METHODS: The co-culture model was constructed by co-culturing K562 cells with bone marrow stromal cells isolated and cultured from the patients with chronic myelogenous leukemia. The IC50 values of K562 cells exposed to imatinib were quantified by MTT assay. The apoptotic rates of K562 cells exposed to 0.5 μmol/L imatinib for 72 hours were detected by flow cytometry through Annexin V-FIT/PI labeling. The cell cycles, cell cycle protein (cyclin A, cyclin D1 and cyclin E) expression of K562 cells co-cultured with bone marrow stromal cells for 72 hours were analyzed by flow cytometry.
RESULTS AND CONCLUSION: The IC50 values of co-culture group and suspension culture group were respectively (0.52±0.02) μmol/L and (1.27±0.05) μmol/L, and their comparison showed significant differences (P < 0.01). After 72 hours of treatment with 0.5 μmol/L imatinib, the apoptotic rates in the co-culture group and suspension culture group were respectively (15.48±4.17)% and (32.01±6.83)%, and their comparison showed significant differences (P < 0.01). The percentages of G₀-G₁phase of K562 cells co-cultured with bone marrow stromal cells for 72 hours were (48.81±8.27)%, which were significantly higher than the suspension culture group (25.78±3.26%) (P < 0.01). The co-culture with bone marrow stromal cells from the patients with chronic myelogenous leukemia could mediate K562 cells resistance to imatinib. The mechanism was possibly related with G₀/G₁arrest of K562 cells induced by co-culture with bone marrow stromal cells.

中国组织工程研究杂志出版内容重点：干细胞；骨髓干细胞；造血干细胞；脂肪干细胞；肿瘤干细胞；胚胎干细胞；脐带脐血干细胞；干细胞诱导；干细胞分化；组织工程

全文链接：

Key words: leukemia, myelogenous, chronic, BCR-ABL positive, bone marrow cells, K562 cells, cell cycle

CLC Number:

R394.2

Wang Ji-gang, Zhou Fan, Liu Yan-qin, Bai Ying, Liu Jing-hua, Wu Dan-tong. Influences of co-culture with primary bone marrow stromal cells on imatinib sensitivity and cell cycles of K562 cells [J]. Chinese Journal of Tissue Engineering Research, 2014, 18(28): 4450-4454.

Figures/Tables 3

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