多发性骨髓瘤中miR-21与SPRY2基因表达的相关性研究

The research of the correlation of miR-21 with SPRY2 expression in multiple myeloma

目的检测多发性骨髓瘤(MM)患者外周血miR-21的表达水平,研究MM中miR-21与SPRY2基因表达的相关性。方法选择30例MM患者、15例意义未明单克隆丙种球蛋白病(MGUS)患者及20例正常对照(NC)的门诊患者,用实时荧光PCR定量检测miR-21和SPRY2表达水平;Western blot检测miR-21和SPRY2在MM细胞系中表达;在荧光显微镜下观察:U266用脂质体转染荧光标记的miR-21mimic/inhibitor后miR-21的表达。结果 MM组血清循环miR-21的表达水平较MGUS组和NC组显著增高,差异有统计学意义(P<0.01);在miR-21内源性高表达的MM细胞株中,SPRY2明显低表达;反之,明显高表达;荧光标记的miR-21 mimic/inhibitor,转染效率90%以上,转染mimics细胞miR-21表达量(98.6±14.2)较未处理的U266细胞miR-21表达量(0.82±0.13)升高了120.2倍(差异有显著的统计学意义,P<0.001),转染inhibitor细胞miR-21表达量(0.37±0.06)较未处理细胞降低了61.9%(差异有明显的统计学意义,P<0.05)。结论 miR-21可能是导致SPRY2在MM中表达下调的一个负性调控因子。miR-21与MM的发生发展和疾病预后有密切关系,可作为判断MM患者预后不良指标之一。

Objective To detect the level of miR-21 expression in peripheral blood of multiple myeloma （MM） patients and to study the correlation between miR-21 and SPRY2 expression.Methods30 MM patients, 15 monoclonal gammopathy of undetermined significance（MGUS） patients and 20 normal control （NC） participants were chosen and their miR-21 and SPRY2 expression were detected by real-time fluorescent quantitative PCR. The miR-21 and SPRY2 expression in the U266 MM cell line was detected using Western blot. The miR-21 mimic/inhibitor was tagged by fluorescent and transfected into U266 cells using liposomal transfection method, and the miR-21 expression in both transfected and untreated U266 cells were then observed using fluorescence microscope.Results The serum circulating miR-21 level in MM patients was significantly higher than in MGUS patients and NC participants （P〈0.01）. In the MM cell lines where the miR-21 was endogenously high-expressed, the SPRY2 expression was low, and vice versa. The transfection efficiency of the fluorescent-tagged miR-21 mimic/inhibitor into U266 cells was over 90%. The miR-21 expression level in mimic-transfected U266 cells（98.6±14.2） was over 120 times higher than in untreated U266 cells（0.82±0.13）（P〈0.001）. On the other hand, the miR-21 expression level in inhibitor-transfected U266 cells（98.6±14.2） was 61.9% lower than in untreated U266 cells（0.82±0.13）（P〈0.05）.ConclusionMiR-21 may contribute to the down-regulation of SPRY2 expression in MM patients and may involve in the development and prognosis of MM. Therefore, miR-21 becomes a potential prognostic indicator for MM patients.