去甲基斑蝥素促进2-氰基-3，12-二氧代齐墩果烷-1，9（11）-二烯-28-酸甲酯介导的抗骨肉瘤作用  

NCTD promotes C - 28 methyl ester of 2 - cyano - 3,12 - dioxoolen - 1,9 - dien - 28 - oic acid （ CDDO - Me） - mediated anti - osteosarcoma in osteosarcoma cells

作　　者：张翼[1] 时程程 张华鹏[3] 王海涛[1] 殷力[1] Zhang Yi;Shi Chengcheng;Zhang Huapeng;Wang Haitao;Yin Li(Department of Orthopaedics, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Chin;Department of Pharmacy, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China;Department of Hepatobiliary and Pancreatic Surgery, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Chin)

机构地区：[1]郑州大学第一附属医院骨科,450052 [2]郑州大学第一附属医院药学部,450052 [3]郑州大学第一附属医院肝胆胰外科,450052

出　　处：《中华实验外科杂志》2018年第6期1115-1117,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金青年科学基金（81702663）;河南省医学科技攻关计划项目（201701001）;河南省科技厅科技计划项目（162300410094）;河南省教育厅：河南省高等学校重点科研项目（17A310011、18A320049）;郑州大学第一附属医院院内青年基金项目（YNQN2015163、YNQN2017044）

摘　　要：目的观察去甲基斑蝥素（NCTD）促进2-氰基-3，12-二氧代齐墩果烷-1，9（11）-二烯．28-酸甲酯（CDDO-Me）介导的抗骨肉瘤作用，探讨相关分子机制。方法采用NCTD单药、CDDO-Me单药或者两药联合处理U-20S和Saos-2骨肉瘤细胞株，通过细胞计数试剂盒（CCK-8）细胞活性检测、锥虫蓝染色计数检测细胞死亡率，检测NCTD和CDDO-Me的联合抗骨肉瘤效应；通过Westernblot实验，研究两药联合抗骨肉瘤作用的分子机制。结果CCK-8结果显示，在U-20S细胞株中，CDDO-Me的半数抑制剂量（IC50）值为1．1μmol／L，与3、10和30μmol／L的NCTD联合处理后，Ic50值分别为0．70、0．30和0．03txmol／L，分别降低了37％、81％和99％；在Saos-2细胞株中观察到NCTD对CDDO-Me介导的细胞抑制作用也具有明显的增强作用。10μmol／L的NCTD单药、0．5μmol／L的CDDO-Me单药以及两者联合处理U一20S和Saos-2细胞24h，锥虫蓝染色计数检测显示在U-20S细胞株中，NCTD和CDDO-Me单药分别诱导25％和36％的细胞死亡，而两者联合则能够诱导79％左右的细胞死亡，比单药均具有极为显著的增强作用（P=0．000）；在Saos-2细胞株中，NCTD和CDDO-Me单药分别诱导19％和43％的细胞死亡，而两者联合则能够诱导82％左右的细胞发生死亡，也比单药具有极为显著的增强作用（P=0．000）。Westernblot结果显示在U-20S和Saos-2细胞株中，NCTD或CDDO-Me联合应用能够激发半胱氨酰天冬氨酸特异性蛋白酶（Caspase）-3高度活化，导致聚腺苷二磷酸核糖聚合酶（PARP）绝大部分裂解，促进了凋亡信号的活化。结论NCTD通过上调凋亡信号能够促进CDDO-Me对骨肉瘤细胞的杀伤作用，促进CDDO-Me介导的抗骨肉瘤作用。Objective To explore whether demethylcantharidin （NCTD） could promote C -28 methyl ester of 2 - eyano - 3,12 - dioxoolen - 1,9 - dien - 28 - oic acid （ CDDO - Me） - mediated anti - osteosarcoma and related molecular mechanisms. Methods U -20S and Saos -2 osteosarcoma cells were treated with single or dual drugs. Viability of cells was measured by cell counting kit - 8 （ CCK - 8 ）, and trypan blue staining count was used to measure the cell mortality. The anti - osteosarcoma effects of NCTD combined with CDDO -M were tested, and the molecular mechanisms were studied by Western blotting. Results The results of CCK - 8 showed that the half maximal inhibitory concentration （ IC50 ） value of CDDO - Me was 1.1 μmol/L in U -20S cells, and the ICs0 values were 0. 70, 0. 30, and 0. 03 μmol/L after treatment with 3, 10, and 30 μmoL/L NCTD, decreased by 37% , 81% , and 99% , respectively. In the Saos - 2 ceils, it was also observed that NCTD had a significant enhancement effect on CDDO - Me - mediated cell inhibition. After treatment of U -2OS and Saos -2 cells with 10 μmol/L NCTD single drug, 0. 5 μmoL/L CDDO - Me single drug, or combination for 24 h, the results of trypan blue staining counts showed that in U -2OS cells, NCTD and CDDO -Me alone induced 25% and 36% cell death respective- ly, and the combination of NCTD and CDDO- Me could induce about 79% cell death, which was signifi- cantly more potent than single drug （P = 0. 000）. In the Saos - 2 cells, NCTD and CDDO - Me induced 19% and 43% of cell death respectively, while the combination of NCTD and CDDO - Me resulted in ap- proximately 82% of cell death, which was also significantly more potent than monotherapy （ P = 0. 000）. Western blotting results showed that in U - 2OS and Saos - 2 cells, the combination of NCTD and CDDO - Me could stimulate the activation of cysteinyl aspartate - specific protease （Caspase） - 3, resul- ting in the majority of poly adenosine diphosphate -ribose polymerase （PARP） cleavage, and promote the acti

关 键 词：骨肉瘤 2-氰基-3 12-二氧代齐墩果烷-1 9(11)-二烯-28-酸甲酯 去甲基斑蝥素 脱噬作用 

分 类 号：R738.1[医药卫生—肿瘤]