Kim Si Hyoung, Kang Jun Goo, Kim Chul Sik, Ihm Sung-Hee, Choi Moon Gi, Yoo Hyung Joon, Lee Seong Jin
1 Division of Endocrinology and Metabolism, Department of Internal Medicine, College of Medicine, Hallym University, Chuncheon, Republic of Korea.
2 Division of Endocrinology and Metabolism, Department of Internal Medicine, Hallym University Sacred Heart Hospital, Anyang-Si, Republic of Korea.
Tumour Biol. 2017 Feb;39(2):1010428317692252. doi: 10.1177/1010428317692252.
In this study, the combined effect of doxorubicin with cucurbitacin B on survival of anaplastic thyroid carcinoma cells was evaluated. For experiments, 8505C and CAL62 human anaplastic thyroid carcinoma cells were used. Cell viability, the percentage of viable cells, and cytotoxic activity were measured using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay, multiplexed cytotoxicity assay, and cytotoxicity assay, respectively. Reactive oxygen species production was measured. In experiments, doxorubicin and cucurbitacin B reduced cell viability in a dose- and time-dependent manner. Cotreatment of doxorubicin and cucurbitacin B, compared with treatment of doxorubicin alone, decreased the percentage of viable cells and increased cytotoxic activity. All of the combination index values were lower than 1.0, suggesting the synergism between doxorubicin and cucurbitacin B in induction of cytotoxicity. In cells treated with both doxorubicin and cucurbitacin B, compared with doxorubicin alone, the protein levels of cleaved poly(adenosine diphosphate-ribose) polymerase and cyclooxygenase 2 and reactive oxygen species production were enhanced. In contrast, the protein levels of B-cell chronic lymphocytic leukemia/lymphoma 2 and survivin and B-cell chronic lymphocytic leukemia/lymphoma 2/B-cell chronic lymphocytic leukemia/lymphoma 2-associated x protein ratio were diminished. The protein levels of Janus kinase 2 and signal transducer and activator of transcription 3 were reduced, while phospho-extracellular signal-regulated kinase 1/2 protein levels were elevated without change in total extracellular signal-regulated kinase 1/2 protein levels. These results suggest that doxorubicin synergizes with cucurbitacin B in induction of cytotoxicity in anaplastic thyroid carcinoma cells. Moreover, synergistic cytotoxicity of doxorubicin with cucurbitacin B is mediated by B-cell chronic lymphocytic leukemia/lymphoma 2 family proteins, survivin, and reactive oxygen species and modulated by Janus kinase 2/signal transducer and activator of transcription 3 and extracellular signal-regulated kinase 1/2 in anaplastic thyroid carcinoma cells.
在本研究中，评估了阿霉素与葫芦素B联合作用对间变性甲状腺癌细胞存活率的影响。实验采用8505C和CAL62人间变性甲状腺癌细胞。分别使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法、多重细胞毒性测定法和细胞毒性测定法测量细胞活力、活细胞百分比和细胞毒性活性。检测了活性氧的产生。实验中，阿霉素和葫芦素B以剂量和时间依赖性方式降低细胞活力。与单独使用阿霉素相比，阿霉素与葫芦素B联合处理降低了活细胞百分比并增加了细胞毒性活性。所有联合指数值均低于1.0，表明阿霉素与葫芦素B在诱导细胞毒性方面具有协同作用。在同时用阿霉素和葫芦素B处理的细胞中，与单独使用阿霉素相比，裂解的聚（腺苷二磷酸核糖）聚合酶和环氧化酶2的蛋白水平以及活性氧的产生增加。相反，B细胞慢性淋巴细胞白血病/淋巴瘤2和存活素的蛋白水平以及B细胞慢性淋巴细胞白血病/淋巴瘤2/B细胞慢性淋巴细胞白血病/淋巴瘤2相关X蛋白比值降低。Janus激酶2和信号转导及转录激活因子3的蛋白水平降低，而磷酸化细胞外信号调节激酶1/2蛋白水平升高，细胞外信号调节激酶1/2总蛋白水平无变化。这些结果表明，阿霉素与葫芦素B在诱导间变性甲状腺癌细胞毒性方面具有协同作用。此外，阿霉素与葫芦素B的协同细胞毒性由B细胞慢性淋巴细胞白血病/淋巴瘤2家族蛋白、存活素和活性氧介导，并由Janus激酶2/信号转导及转录激活因子3和细胞外信号调节激酶1/2在间变性甲状腺癌细胞中调节。
