靶向MTH1抑制多发性骨髓瘤细胞增殖和诱导凋亡的作用研究  

Proliferation-Inhibitory and Apoptosis-Inducing Effects of Targeting MTH1 on Multiple Myeloma Cells

作　　者：王欣[1] 吴颖[1] 张林[2,3] 李宇翔 韩冰 史雪 王伟[1] 王立生[2,3] WANG Xin;WU Ying;ZHANG Lin;LI Yu-Xiang;HAN Bing;SHI Xue;WANG Wei;WANG Li-Sheng(Department of Hematology,Affiliated Hospital of Qingdao University,Qingdao 266000,Shandong Province,China;Molecular Diagnosis and Regenerative Medicine Laboratory,Affiliated Hospital of Qingdao University,Qingdao 266000,Shandong Province,China;Institute of Radiation Medicine,Academy of Military Medical Sciences,Academy of Military Sciences,Beijing 100850,China;College of Nursing,Jilin University,Jilin Province,China)

机构地区：[1]青岛大学附属医院血液科,山东青岛266000 [2]青岛大学附属医院分子诊断与再生医学实验室,山东青岛266000 [3]军事科学院军事医学研究院放射医学研究所,北京100850 [4]吉林大学护理学院,吉林长春130021

出　　处：《中国实验血液学杂志》2020年第5期1598-1604,共7页Journal of Experimental Hematology

摘　　要：目的:检测MTH1在多发性骨髓瘤(MM)患者CD138阴性细胞(CD138^-)及CD138阳性细胞(CD138^+)中的表达,探讨MTH1抑制剂对MM细胞株U266细胞的形态、增殖和凋亡等细胞生物学特性的影响。方法:分离MM患者CD138^-及CD138^+并提取RNA,应用Q-PCR分别检测NUDT家族的表达情况。利用MM细胞系U266观察白细胞介素-6(IL-6)对MTH1表达变化的影响。在荧光显微镜下观察TH588作用于U266细胞48 h后的形态变化,用DAPI染色法观察细胞核的变化。应用荧光素酶活性检测TH588对U266细胞增殖的影响,应用流式细胞术检测TH588对U266细胞凋亡的影响。结果:MM患者CD138^+中MTH1、NUDT2、NUDT5及NUDT21表达较CD138^-高(P<0.05)。荧光素酶报告显示TH588处理U266细胞后,与对照组相比,荧光强度值明显降低,且药物浓度越大,荧光强度值越低(r=-0.91);应用IL-6能使MTH1表达水平增高,荧光显微镜下观察,TH588处理U266细胞48 h后,细胞出现皱缩、变小以及形状不规则等表现,DAPI染色后,TH588处理过的细胞呈现细胞核缩小、染色不均、花瓣状和放射状等凋亡特征;流式细胞术分析结果显示,TH588处理U266细胞48 h后,存活细胞比例明显降低(P<0.05)。结论:MTH1在部分MM患者的CD138^+中高表达,U266细胞在IL-6的作用下可见MTH1表达增高,MTH1抑制剂TH588对MM U266细胞株具有明显地抑制增殖和诱导细胞凋亡的作用。Objective:To detect the expression of MutT homolog 1(MTH1)in CD138-negative cells(CD138^-)and CD138-positive cells(CD138^+)cells of the patients with multiple myeloma(MM),and to explore the effect of MTH1 inhibitor TH588 on cell morphology,proliferation and apoptosis of MM cell U266.Methods:CD138-and CD138^+cells of MM patients were isolated,and RNA was extracted.The expression of NUDT family was detected by Q-PCR.MM cell line U266 was used to observe the effect of IL-6 on MTH1 expression.Using fluorescence microscopy to observe the morphological changes of U266 after treatment by TH588 for 48 hours.DAPI staining was used to investigate the nuclear change.Luciferase was used to detect the effect of TH588 on U266 cell proliferation.After treatment with TH588 for 48 h,the change of apoptosis in MM U266 cells was detected by flow cytometer.Results:In some MM patients,the expression of MTH1,NUDT2,NUDT5 and NUDT21 in CD138^+cells was higher than that in CD138^-(P<0.05).The luciferase report showed that after treatment of U266 cells with TH588,the fluorescence intensity was significantly lower than that of the control group,and the fluorescence intensity decreased with the increase of drug concentration(r=-0.91).IL-6 could increase the expression of MTH1.Fluorescence microscopy showed that after TH588 treatment of U266 cells for 48 hours,the cells appeared shrinking,smaller,and irregular in shape.After DAPI staining,the TH588-treated cells showed apoptosis characteristics,such as nuclear shrinkage,uneven staining,petal,and radial shape.Flow cytometry showed that the proportion of U266 viable cells decreased significantly after treatment with TH588 for 48 h(P<0.05).Conclusion:MTH1 highly expresses in CD138^+cells of some MM patients.MTH1 expression can increase in U266 cells treated by IL-6.The MTH1 inhibitor TH588 possesses proliferation-inhibitory effect and apoptosis-inducing effect on MM cell U266.

关 键 词：MTH1 NUDT TH588 多发性骨髓瘤 

分 类 号：R733.3[医药卫生—肿瘤]