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目的 从阻滞糖代谢途径,探讨三溴丙酮酸(3-BrPA)抑制恶性转化的树突状细胞(DC)增殖的可能性.方法 采用髓源性小鼠DC与人脑胶质瘤干细胞(SU3细胞株)体外共培养的方法,诱导DC恶性转化,对恶性转化的DC单克隆,培养的细胞命名为胶质瘤干细胞诱导恶性转化的DC (GSPC-MTDCs).采用CCK-8法和细胞平板克隆形成实验检测SU3、正常DC、GSPC-MTDCs的增殖能力;采用Transwell小室侵袭实验检测细胞的侵袭能力.将GSPC-MTDCs接种于20只无荧光裸小鼠皮下,接种后11d,选取致瘤均一的10只裸鼠分为干预组和对照组,每组5只.干预组给予3-BrPA(5 mg·kg-1·d-1)腹腔注射,对照组给予等体积的生理盐水腹腔注射,共干预24 d.比较两组的致瘤情况.结果 (1) GSPC-MTDCs具有永生化特征,与SU3相比,其增殖速度加快,克隆形成率明显增高[(38.1±1.9)％对比(30.2±2.7)％,P＜0.01];Transwell小室侵袭实验显示,GSPC-MTDCs侵袭能力强于SU3[(1 042±49)个对比(782 ±38)个,P＜0.01)].(2)GSPC-MTDCs接种于无荧光裸小鼠皮下和尾静脉后全部致瘤.3-BrPA干预后24 d,干预组和对照组肿瘤的重量分别为(1.5±0.6)g、(3.3±1.0)g,差异有统计学意义(P =0.0107).病理学观察显示,相较于干预组,对照组细胞核异形明显、核分裂象多、血管丰富、炎症细胞浸润明显、c-myc阳性细胞比例高.结论 胶质瘤干细胞具有诱导DC恶变的潜能;3-BrPA具有抑制GSPC-MTDCs增值的作用.

Objective To explore the possible inhibiting effect of 3-bromopyruvic acid (3-BrPA)on proliferation of malignant transformed dendritic cells by blocking the pathway of glucose metabolism.Methods In vitro,co-cultured mice myeloid dendritic cells (DC) with human glioma stem ceils (SU3)were used to induce the malignant transformation of DC.Among DC monoclonal for malignant transformation,i.e.,glioma stern/progenitor cells induced by malignant transformation of dendritic cells (GSPC-MTDCs),proliferative abilities of SU3,normal DC and GSPC-MTDCs were detected by CCK-8 and flat clone formation assay.Cell invasiveness was detected by Transwell chamber invasion assay.Subcutaneous inoculation of GSPC-MTDCs was performed in 20 non-fluorescent nude mice.At 11 d post inoculation,10 nude mice with uniform tumorigenesis were divided into intervention group and control group with 5 mice in each group.The intervention group was given 3-BrPA (5 mg · kg-1 · d-1) intraperitoneal treatment,while the control group was given the same volume of normal saline.After 24 days of intervention,the tumorigenesis was compared between 2 groups.Results (1) GSPC-MTDCs had immortalized characteristics.Compared with SU3,the proliferation rate of GSPC-MTDCs was faster and the clone formation rate was significantly higher (38.1 ± 1.9％ vs.30.2 ± 2.7％,P ＜ 0.01).Transwell chamber invasion test showed that GSPC-MTDCs had stronger invasive ability than SU3 (1 042 ± 49 vs.782 ± 38,P ＜ 0.01).(2) GSPC-MTDCs inoculated into the subcutaneous and caudal veins of non-fluorescent nude mice all formed tumors.At 24 days after 3-BrPA intervention,the tumor masses in the intervention group and the control group were 1.5 ± 0.6 g and 3.3 ± 1.0 g,respectively.The difference was statistically significant (P =0.0107).Pathological observation showed that compared with the intervention group,the control group had more abnormal nuclei,more mitosis,abundant blood vessels,obvious infiltration of inflammatory cells and higher proportion of cmyc positive cells.Conclusions Glioma stem cells have the potential to induce malignant transformation of DC cells.3-BrPA could inhibit the proliferation of GSPC-MTDCs.