Department of Anatomy, Harbin Medical University, Harbin, China.
Anat Rec (Hoboken). 2012 Aug;295(8):1260-6. doi: 10.1002/ar.22497. Epub 2012 May 23.
Bladder cancer is the most common neoplasm in the urinary system. This study assesses arctigenin anti-tumor activity in human bladder cancer T24 cells in vitro and the underlying molecular events. The flow cytometry analysis was used to detect cell-cycle distribution and apoptosis. Western blotting was used to detect changes in protein expression. The data showed that arctigenin treatment reduced viability of bladder cancer T24 cells in a dose- and time-dependent manner after treatment with arctigenin (10, 20, 40, 80, and 100 μmol/L) for 24 hr and 48 hr. Arctigenin treatment clearly arrested tumor cells in the G1 phase of the cell cycle. Apoptosis was detected by hoechst stain and flow cytometry after Annexin-V-FITC/PI double staining. Early and late apoptotic cells were accounted for 2.32-7.01% and 3.07-7.35%, respectively. At the molecular level, arctigenin treatment decreased cyclin D1 expression, whereas CDK4 and CDK6 expression levels were unaffected. Moreover, arctigenin selectively altered the phosphorylation of members of the MAPK superfamily, decreasing phosphorylation of ERK1/2 and activated phosphorylation of p38 significantly in a dose-dependent manner. These results suggest that arctigenin may inhibit cell viability and induce apoptosis by direct activation of the mitochondrial pathway, and the mitogen-activated protein kinase pathway may play an important role in the anti-tumor effect of arctigenin. The data from the current study demonstrate the usefulness of arctigenin in bladder cancer T24 cells, which should further be evaluated in vivo before translation into clinical trials for the chemoprevention of bladder cancer.
膀胱癌是泌尿系统最常见的肿瘤。本研究评估了牛蒡子苷元在体外人膀胱癌 T24 细胞中的抗肿瘤活性及其潜在的分子事件。采用流式细胞术分析检测细胞周期分布和细胞凋亡。采用 Western blot 检测蛋白表达变化。结果显示，牛蒡子苷元处理后，膀胱癌 T24 细胞活力在浓度和时间依赖性下降，处理浓度分别为 10、20、40、80 和 100μmol/L，作用时间分别为 24h 和 48h。牛蒡子苷元处理明显将肿瘤细胞阻滞于细胞周期 G1 期。用 Hoechst 染色和 Annexin-V-FITC/PI 双染后流式细胞术检测到凋亡。早期和晚期凋亡细胞分别占 2.32%-7.01%和 3.07%-7.35%。在分子水平上，牛蒡子苷元处理降低了细胞周期蛋白 D1 的表达，而 CDK4 和 CDK6 的表达水平不受影响。此外，牛蒡子苷元选择性地改变了 MAPK 超家族成员的磷酸化，ERK1/2 的磷酸化明显减少，p38 的激活磷酸化显著增加，呈剂量依赖性。这些结果表明，牛蒡子苷元可能通过直接激活线粒体途径抑制细胞活力并诱导细胞凋亡，丝裂原激活蛋白激酶途径可能在牛蒡子苷元的抗肿瘤作用中发挥重要作用。本研究的数据表明牛蒡子苷元在膀胱癌 T24 细胞中的有效性，在将其转化为膀胱癌化学预防的临床试验之前，应进一步在体内进行评估。
