摘要
目的探讨FMS样的酪氨酸激酶3(FLT3)内部串联重复(ITD)突变(FLT3-ITD)在多发性骨髓瘤中的表达及对信号调节蛋白Notch的影响。方法选择2014年7月至2017年8月自愿接受治疗的初发多发性骨髓瘤患者64例设为观察组;选择同期接受治疗的白细胞、血小板略低但骨髓检查未见明显异常者64例设为对照组。采集骨髓并分离单个核细胞,采用实时荧光定量聚合酶链式反应(qRT-PCR)法测定骨髓单个核细胞FLT3-ITD mRNA、Notch mRNA表达水平;采用免疫组织化学染色法检测FLT3-ITD、Notch蛋白在骨髓中表达水平。选择骨髓瘤U226细胞,传代培养后,用Lipofectamine 2000转染细胞(阴性对照)及FLT3-ITD/小干扰RNA(siRNA)慢病毒上清液培养(FLT3-ITD组),观察siRNA沉默骨髓瘤U226细胞FLT3-ITD突变对Notch mRNA表达的影响。结果观察组骨髓单个核细胞FLT3-ITD、Notch mRNA表达水平均高于对照组(P <0. 01)。免疫组化结果表明,两组患者浆细胞均可见FLT3-ITD、Notch蛋白阳性表达;与对照组相比,观察组中FLT3-ITD阳性细胞比例显著增加[(30. 45±2. 74)%vs (13. 62±1. 92)%,t=12. 066,P <0. 05],而Notch阳性细胞比例显著减少[(10. 77±2. 15)%vs (92. 08±3. 36)%,t=20. 307,P <0. 05]。siRNA沉默骨髓瘤U226细胞致FLT3-ITD突变后Notch mRNA表达水平下降(P <0. 01)。结论 FLT3-ITD在多发性骨髓瘤中呈高表达,并可调节Notch信号调节蛋白mRNA表达水平,可能与多发性骨髓瘤的发生、发展关系密切。
Objective To investigate the expression of FMS-like tyrosine kinase-3（FLT-3）-internal tandem duplication（ITD） mutation（FLT3-ITD） in multiple myeloma and its impact on the level of signal regulating protein Notch mRNA.Methods Sixty-four patients with primary multiple myeloma voluntarily received treatment between July 2014 and August2017 were selected as research objects（observation group）,and 64 patients with slightly lower white blood cells count and platelets count but without obvious abnormalities in bone marrow examination at the same period were selected as control group. The bone marrow were collected and the mononuclear cells were isolated from bone marrow. Real-time fluorescence quantitative polymerase chain reaction（qRT-PCR） was used to detect the expressions of FLT3-ITD and Notch mRNAs in bone marrow mononuclear cells. Immunohistochemical staining method was used to detect the expressions of FLT3-ITD and Notch proteins in bone marrow. After subculture,the myeloma U226 cells were transfected with Lipofectamine 2000（negative control group） or with lentivirus FLT3-ITD/siRNA（small interfering RNA）supernatant fluid（FLT3-ITD group）to observe the influence of siRNA silencing of FLT3-ITD on Notch mRNA expression in myeloma U226 cells. Results FLT3-ITD and Notch mRNAs expression levels in bone marrow mononuclear cells in observation group were significantly higher than those in control group（all P〈0. 01）. Immunohistochemical staining showed that the positive expressions of FLT3-ITD and Notch proteins were found in plasma cells of bone marrow specimens in both two groups. Compared with control group,the proportion of cells with FLT3-ITD protein positive expression increased significantly[（30. 45 ± 2. 74） %vs（13. 62 ± 1. 92） %,t = 12. 066,P〈0. 05],while the proportion of cells with Notch protein positive expression decreased significantly[（10. 77 ± 2. 15） % vs（92. 08 ± 3. 36） %,t = 20. 307,P〈0. 05] in observation group. After silencing FLT3-ITD with siR
作者
刘涛
赵一
LIU Tao;ZHAO Yi(Department of Pathology,Yunnan Traditional Chinese Medicine Hospital,Kunming,Yunnan 650021,China)
出处
《中国临床研究》
CAS
2018年第11期1485-1488,共4页
Chinese Journal of Clinical Research
