骨髓微环境对多发性骨髓瘤细胞microRNA-21和microRNA-30b表达的影响

microRNA-21 and microRNA-30b expression in multiple myeloma

目的研究骨髓微环境对多发性骨髓瘤（MM）细胞microRNA（miR）-21和miR-30b的调变作用。方法培养破骨细胞，采用MACS法分选纯化CD138^＋骨髓瘤细胞。采用实时定量PCR的方法检测MM患者瘤细胞和细胞株以及与破骨细胞共培养的MM患者瘤细胞miR-21和miR-30b的表达量。结果MM患者的外周血单个核细胞经破骨细胞培养基培养后10～14d形成破骨细胞，瘤细胞与破骨细胞共培养组和未与破骨细胞共培养组相比较，细胞活力明显增高，miR-21的表达量增高了1．3～5．9倍，miR-30b表达量降低27．5％～76．9％。在正常浆细胞、MM患者瘤细胞和MM细胞株，miR-21表达水平分别为1．9±0．8、6．5±4．9和35．1±36．2，而miR-30表达分别为13．6±1．8、7．2±6．3和4．5±1．9。结论骨髓微环境可调控miR-21和miR-30b的表达，miR-21起癌基因作用，而miR-30b起抑癌基因作用。

Objective To investigate the expression of miR-21 and miR-30b in muhiple myeloma （MM）. Methods Peripheral blood mononuelear cells from patients with MM were cultured at 2.5 × 10^6 cells/ml in α-MEM supplemented with 10% of fetal bovine serum, antibiotics, RANKL （50 ng/ml） , and macrophage colony-stimulating factor （25 ng/ml） for 10 to 14 days to obtain osteoclasts with bone-resorbing activity. Primary myeloma cells were purified from 12 MM patients. Of them, 8 samples were cocuhured with osteoclasts and 4 as noncoeuhured control. The expression of miR-21 and miR-30b was detected by real-time PCR. Results The viability of MM cells recovered from cocuhures was higher than those of noneocuhured control. After eocuhured with osteoclasts, primary myeloma cells from eight patients exhibited a 2.3 - 6.9- fold increase in miR-21 expression and 1.38 - 4.32-fold decrease in miR-30b expression compared with controis. In highly purified plasma cells from 3 healthy subjects, 12 MM patients and 11 MM cell lines, the expression of miR-21 was 1.9 ± 0.8, 6.5± 4.9 and 35.1 ± 36.2, respectively ; the expression of miR-30b was 13.6 ± 1.8, 7.2 ± 6.3 and 4.5 ±1.9, respectively. Conclusions miR-21 acts as an oncogene and miR- 30b a tumor suppressor gene in MM.