摘要: 目的:观察粒细胞集落刺激因子(GCSF)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、GCSF单药组(1000、2000 U/ml)、ATRA单药组(10 μmol/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,AnnexinV/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<005),联合用药组生长抑制率均大于单药组(P<005);而在U266细胞这种作用不明显(P>005)。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<005),对照组与GCSF单药组RARα2表达无明显差别(均P>005)。U226细胞的对照组、GCSF单药组不表达RARα2,ATRA单药组及GCSF+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。
关键词 :
多发性骨髓瘤/药物疗法;多发性骨髓瘤病理学;粒细胞集落刺激因子 ,
维甲酸/治疗应用;细胞凋亡;细胞分化
Abstract :Objective: To investigate the effect of alltrans retinoid acid (ATRA) and granulocyte colonystimulating factor (GCSF)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 GCSF. The cells were divided into 6 groups: control groups, GCSF groups (treated with 1000 U/ml and 2000 U/ml), ATRA groups (treated with 10 μmol/L ATRA) and combined groups (treated with 1000 U/mL or 2000 U/mL GCSF plus 10 μmol/L ATRA). The cell viability, growth and apoptosis were examined by MTT method, inverted microscopy and AnnexinV/PI staining, respectively; RARα2 expression was detected by reverse transcription PCR; morphology change was evaluated by WrightGiemsa staining; CD49e expression were analyzed by flow cytometry. Results: The proliferation of OPM2 cells was inhibited by ATRA treatment(P<005). The growth inhibition rates in combined groups were higher than corresponding single ATRA groups (P<005). However, the above effects in U266 cells were not significant (P >005). The OPM2 cell stained by WrightGiemsa 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 CD49e 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<005); and there was no significant difference between control group and GCSF groups (P>005). Expression of RARα2 in U266 cells of control group and GCSF groups was not detected; and ATRA groups and combination groups had weak expression. Conclusion: ATRA can induce proliferation inhibition in RARα2exprssing myeloma cells, and it may also play a certain role in promoting differentiation of RARα2 positive myeloma cells.
Key words :
Multiple myeloma/drug therapy
Multiple myeloma/pathology
Granulocyte colonystimulating factor
Tretinoin/therapeutic use;
收稿日期: 2013-10-30
基金资助: 湖南省科技厅创新基金(2009SK3185)
通讯作者:
贺艳娟(1965-),女,博士,副教授,硕士生导师,主要从事血液系统恶性肿瘤的临床诊疗及耐药机制研究;Email:Yanjuanh@yahoo.com
Corresponding author:
HE Yanjuan, Email: Yanjuanh@yahoo.com
作者简介 : 蒋端凤(1986-),男,硕士,住院医师,主要从事血液系统疾病的临床诊疗及发病机制研究;Email∶duanfeng989949@126.com