Wang Zhao-Xia, Yang Zhi-Min, Zou Ya-Wei, Li Min-Min, Chen Fu-Xiong, Zhong Guo-Yu, Guan Jing-Ming, Wei Feng-Gui, Wu Shang-Zhi, He Zheng-Tao, Wu Zi-Liang
Department of Pediatrics, Wendeng Central Hospital, Weihai 264400, Shandong Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Feb;19(1):19-23.
The aim of study was to investigate the effect of acute lymphoblastic leukemia (ALL) children bone marrow mesenchymal stem cells (MSC) on resistance of K562/A02 cells and its mechanism. MSC obtained from bone marrow of AL children were cultured and identified. The co-culture of MSC and K562/A02 and the culture of K562/A02 cell suspension alone was performed, of which 2 kinds of cells were treated with same concentration of adriamycin (ADM), and the rate of apoptosis was detected by flow cytometry, bcl-2 and bax of K562/A02 were detected by RT-PCR, while mdr1 gene level was detected by FQ-PCR. The results indicated that the MSC separation and proliferation were viable and steady. The apoptosis rate of the K562/A02 cells co-cultured with MSC was 1.97 ± 0.11%, while apoptosis rate of the K562/A02 cells cultured alone was 8.38 ± 0.29%, there was significant difference (p < 0.05). As compared with the K562/A02 cells cultured alone, the bcl-2 gene expression in K562/A02 cells co-cultured with MSC obviously increased; ratio of bcl-2/bax was obviously enhanced. The mdr1 gene level in K562/A02 co-cultured with MSC was no statistical different from K562/A02 cultured alone (p > 0.05), which suggested that adhesion co-cultured with MSC did not induce mdr1 expression higher than the culture of suspension. It is concluded that the MSC of ALL children can escape the leukemia cells from proapoptotic effect of drugs, the resistance of K562/A02 to ADM may be involved in enhancement of bcl-2 gene expression of K562/A02 cells co-cultured with MSC, but not in relation to mdr1 gene in K562/A02 cells themselves.
本研究旨在探讨急性淋巴细胞白血病（ALL）患儿骨髓间充质干细胞（MSC）对K562/A02细胞耐药性的影响及其机制。从ALL患儿骨髓中获取MSC并进行培养和鉴定。将MSC与K562/A02进行共培养，并单独培养K562/A02细胞悬液，两种细胞均用相同浓度的阿霉素（ADM）处理，采用流式细胞术检测凋亡率，用RT-PCR检测K562/A02的bcl-2和bax，用FQ-PCR检测mdr1基因水平。结果表明，MSC分离及增殖良好且稳定。与MSC共培养的K562/A02细胞凋亡率为1.97±0.11%，而单独培养的K562/A02细胞凋亡率为8.38±0.29%，差异有统计学意义（p<0.05）。与单独培养的K562/A02细胞相比，与MSC共培养的K562/A02细胞中bcl-2基因表达明显增加；bcl-2/bax比值明显升高。与MSC共培养的K562/A02中mdr1基因水平与单独培养的K562/A02相比无统计学差异（p>0.05），提示与MSC黏附共培养未诱导mdr1表达高于悬液培养。结论：ALL患儿的MSC可使白血病细胞逃避药物促凋亡作用，K562/A02对ADM的耐药性可能与MSC共培养的K562/A02细胞bcl-2基因表达增强有关，而与K562/A02细胞自身的mdr1基因无关。
