Odeh Lena Hisham, Talib Wamidh H, Basheti Iman A
Department of Clinical Pharmacy and Therapeutics, Applied Science University, Amman, Jordan.
J Cancer Res Ther. 2018 Jun;14(Supplement):S324-S330. doi: 10.4103/0973-1482.235349.
To test the anticancer potential of a combination of thymoquinone (TQ) and melatonin (MLT) against breast cancer implanted in mice.
The antiproliferative activity of TQ, MLT, and their combination was tested against mouse epithelial breast cancer cell line (EMT6/P) using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. The combination index (CI) was calculated using isobolographic method. Balb/C mice were transplanted with EMT6/P cell line and in vivo antitumor activity was assessed for TQ, MLT, and their combination. Changes in tumor size were measured for each treatment. Histological examination of tumor sections was performed using standard hematoxylin/eosin staining protocol and TUNEL colorimetric assay was used to test the apoptosis induction ability for all treatments. Immunohistochemical staining was used to detect vascular endothelial growth factor (VEGF) expression in tumor section and ELISA was used to measure serum levels of interferon gamma (INF-γ) and interleukin-4. Serum levels of the liver enzymes aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were used as biomarkers of hepatotoxicity of the combination therapy.
Synergistic anticancer effect was observed between TQ and MLT with CI value of 0.552. The combination of TQ and MLT caused a significant decrease in tumor size with a percentage cure of 60%. The combination therapy induced extensive necrosis, increased apoptosis rate, and decreased VEGF expression in tumor sections. Serum levels of INF-γ were increased in mice treated with combination therapy and AST and ALT levels were close to their normal values.
The combination TQ and MLT act synergistically to inhibit breast cancer implanted in mice. The anticancer effect of this combination is mediated by induction of apoptosis, angiogenesis inhibition, and activation of T helper 1 anticancer immune response.
测试百里醌（TQ）与褪黑素（MLT）联合使用对植入小鼠体内的乳腺癌的抗癌潜力。
采用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法检测TQ、MLT及其组合对小鼠上皮性乳腺癌细胞系（EMT6/P）的抗增殖活性。使用等效线法计算联合指数（CI）。将EMT6/P细胞系移植到Balb/C小鼠体内，评估TQ、MLT及其组合的体内抗肿瘤活性。测量每种治疗方法下肿瘤大小的变化。使用标准苏木精/伊红染色方案对肿瘤切片进行组织学检查，并使用TUNEL比色法测试所有治疗方法的凋亡诱导能力。采用免疫组织化学染色检测肿瘤切片中血管内皮生长因子（VEGF）的表达，采用酶联免疫吸附测定法测量血清中干扰素γ（INF-γ）和白细胞介素-4的水平。血清中天冬氨酸转氨酶（AST）和丙氨酸转氨酶（ALT）水平作为联合治疗肝毒性的生物标志物。
TQ与MLT之间观察到协同抗癌作用，CI值为0.552。TQ与MLT联合使用可使肿瘤大小显著减小，治愈率达60%。联合治疗诱导肿瘤切片出现广泛坏死，增加凋亡率，并降低VEGF表达。联合治疗的小鼠血清INF-γ水平升高，AST和ALT水平接近正常。
TQ与MLT联合使用具有协同作用，可抑制植入小鼠体内的乳腺癌。这种联合用药的抗癌作用是通过诱导凋亡、抑制血管生成和激活辅助性T细胞1抗癌免疫反应介导的。
