【摘要】 目的:观察粒细胞集落刺激因子(G-CSF)与全反式维甲酸(ATRA)对骨髓瘤细胞生长、分化及RARα2表达的影响,探讨两药联合用于临床治疗骨髓瘤的可能性。方法:以人骨髓瘤细胞RARα2阳性细胞株(OPM2)和RARα2阴性细胞株(U266)为研究对象,设对照组、G-CSF单药组(1000、2000 U/ml)、ATRA单药组(1.0μmol/L)及两药联合组共6个平行组。采用MTT法检测细胞活力,倒置显微镜动态观察细胞凋亡过程,Annexin-V/PI双染法检测早期凋亡;瑞氏-姬母萨染色观察细胞形态学,流式细胞术分析CD49e表达;逆转录PCR检测RARα2 mRNA表达。结果:ATRA可抑制OPM2细胞的生长(P<0.05),联合用药组生长抑制率均大于单药组(P<0.05);而在U266细胞这种作用不明显(P>0.05)。瑞氏-姬母萨染色显示在含ATRA的处理组中,OPM2细胞有细胞核缩小、染色质聚集浓缩、核仁数量减少、胞浆量增加并呈深蓝色改变;此类改变在U226细胞不明显。U266、OPM2细胞均弱表达CD49e。在OPM2细胞中,联合用药组较对照组、ATRA单药组RARα2表达均增加(P <0.05),对照组与G-CSF单药组RARα2表达无明显差别(均P>0.05)。U226细胞的对照组、G-CSF单药组不表达RARα2,ATRA单药组及G-CSF+ATRA联合组弱表达RARα2。结论:ATRA对RARα2阳性骨髓瘤细胞有抑制细胞增殖作用;ATRA对RARα2阳性骨髓瘤细胞的分化也有一定促进作用。更多还原

【Abstract】 Objective:To investigate the effect of all-trans retinoid acid(ATRA)and granulocyte colony-stimulating factor(G-CSF) on the growth,apoptosis,differentiation and expression of RARα2 of myeloma cells.Methods:Myeloma cell lines OPM2(RARα2 positive) and U266(RARα2 negative) were treated with ATRA in the presence or absence of G-CSF.The cells were divided into 6 groups:control groups,G-CSF groups(treated with 1000 U/ml and 2000 U/ml),ATRA groups(treated with 1.0 μmol/L ATRA) and combined groups(treated with 1000 U/mL or2000 U/mL G-CSF plus 1.0 μmol/L ATRA).The cell viability,growth and apoptosis were examined by MTT method,inverted microscopy and Annexin-V/PI staining,respectively;RARα2 expression was detected by reverse transcription PCR;morphology change was evaluated by Wright-Giemsa staining;CD49 e expression were analyzed by flow cytometry.Results:The proliferation of OPM2 cells was inhibited by ATRA treatment(P <0.05).The growth inhibition rates in combined groups were higher than corresponding single ATRA groups(P <0.05).However,the above effects in U266 cells were not significant(P> 0.05).The OPM2 cell stained by Wright-Giemsa in ATRA groups showed that the cell nucleus became smaller,chromatin condensed,number of nucleolus reduced,the volume of cytoplasm increased and the cytoplasm became dark blue.Expression rates of CD49 e were low in both U266 and OPM2 cells.Expression of RARα2 in OPM2 cells of combination groups were higher than those of control group and corresponding single groups(P <0.05);and there was no significant difference between control group and G-CSF groups(P> 0.05).Expression of RARα2 in U266 cells of control group and G-CSF groups was not detected;and ATRA groups and combination groups had weak expression.Conclusion:ATRA can induce proliferation inhibition in RARα2-exprssing myeloma cells,and it may also play a certain role in promoting differentiation of RARα2 positive myeloma cells.更多还原