急性淋巴细胞白血病骨髓细胞单核细胞趋化蛋白-1的表达及其意义

Expression and function of chemokine CCL2 in the bone marrow cells of patients with acute lymphoblastic leukemia

目的探讨单核细胞趋化蛋白-1（CCL2）在B系急性淋巴细胞白血病（B-ALL）患者骨髓中的表达及其对白血病细胞增殖和黏附的影响。方法提取30例B—ALL患者（试验组）及30例非血液肿瘤患者（对照组）骨髓基质细胞（BMMSC）及骨髓单个核细胞（BMMNC），采用酶联免疫吸附法（ELISA）定量检测细胞培养上清液CCL2的质量浓度，反转录一聚合酶链反应（RT-PCR）检测CCL2基因mRNA水平；四甲基偶氮唑蓝比色（M1Tr）法检测细胞增殖及黏附；流式细胞术进行白血病细胞计数。结果试验组的CCL2蛋白质量浓度[（780．12±129．61）Pg／m1]明显高于对照组[（120．49±25．21）pg／ml]（t=4．96，P=0．00）；BMMSC体外培养E清液的CCL2蛋白质量浓度[（572．38±35．39）pg／m1]明显高于BMMNC[（193．85±15．45）Pg／ml]（t=5．37，P=0．00）。在体外，CCL2不能单独刺激白血病细胞增殖，但在BMMSC＋BMMNC共培养体系中，CCL2能刺激白血病细胞增殖，且随质量浓度增加，其增殖作用增强。CCL2在体外能增强白血病细胞与BMMSC黏附，且与质量浓度呈正相关（r=0．824，P=0．02）。结论B-ALL患者骨髓中CCL2水平增高，主要是由BMMSC分泌，白血病细胞能促进BMMSC分泌CCL2，CCL2在BMMSC环境下能刺激白血病细胞增殖，促进白血病细胞与BMMSC黏附，这些作用与CCL2浓度呈正相关。

Objective To investigate the expression of CCL2 and its effects on the proliferation and adhesiveness on leukemia cells in patients with acute lymphoblastic leukemia （ALL）. Methods The bone marrow mesenchymal cells （BMMSC） and bone marrow mononuclear cells （BMMNC） of 30 ALL patients and 30 healthy controls were studied, and CCL2 level was evaluated by ELISA. CCL2 gene mRNA level in ALL was determined by RT-PCR. The cell proliferation and adhesiveness were detected by MTI＂ assays. The cell counts were measured by flow cytometry. Results The BM plasma levels of CCL2 in patients at diagnosis were significantly higher than that in healthy controls [（780.12±129.61） pg/ml vs （120.49±25.21） pg/ml, t = 4.96, P = 0.00]. Supernatant levels of CCL2 in BMMSC were significantly higher than that of BMMNC [（572.38±35.39） pg/ml vs （193.85±15.45） pg/ml, t = 5.37, P = 0.00] in vitro. CCL2 cannot induce leukemia proliferation alone, but could induce leukemia proliferation in BMMNC and BMMSC co-culture in a dose- and time-dependent manner, CCL2 could increase the leukemia adhesive to the BMMSC compared with control （r = 0.824, P = 0.02）. Conclusion Patients with B type ALL had higher levels of CCL2 which was secreted by BMMSC. The leukemia could induce the BMMSC to secrete CCL2. CCL2 could promote the survival and proliferation of leukemia in the presence of BMMSC and BMMNC, and enhance ALL cells adhesion to BMMSC in a dose-dependent manner.