Shen Peng, Wang Hong-Gang, Li Miao-Miao, Ma Qian-Yun, Zhou Chuan-Wen, Pan Feng, Xie Rui
Department of Gastroenterology, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, Jiangsu 223300, PR China.
Department of Gastroenterology, Huai'an First People's Hospital, Nanjing Medical University, Huai'an, Jiangsu 223300, PR China.
Biomed Pharmacother. 2017 Aug;92:78-85. doi: 10.1016/j.biopha.2017.05.065. Epub 2017 May 19.
Isofraxidin (IF), a natural coumarin compound, has been reported to possess anti-cancer activity in human liver cancer. However, whether IF is involved in the regulation of colorectal cancer tumorigenesis and development has been not well elucidated.
The cell proliferation were assessed by Cell Counting Kit-8 (CCK-8) and colony formation test, respectively. The transwell assays were conducted to estimate cell migration and invasion abilities. Further, cell apoptosis was evaluated by confocal microscopy analysis, flow cytometry detection and TdT-mediated dUTP Nick-End Labeling (TUNEL) method. Western blot were performed to detect the expression of related protein.
Herein, the result indicated that IF remarkably bated cell proliferation in human colorectal cancer cells HT-29 and SW-480 in a dose- and time-dependent manner. In addition, IF treatment showed obvious inhibitory activity to cell colony formation in HT-29 and SW-480 cells. Confocal microscopy analysis and flow cytometry detection revealed that IF dramatically induced cell apoptosis in HT-29 and SW-480 cells compared with the control. And IF markedly decreased the expression of anti-apoptotic protein bcl-2, whereas the expression of pro-apoptotic proteins, including caspase-3, caspase-9 and bax, notably increased in HT-29 and SW-480 cells. Besides, IF blocked Akt pathway via inhibition expression of p-Akt. Furthermore, MK2206, an Akt inhibitor, could inhibit cell colony formation and induced apoptosis. This effect is even more obvious in the presence of MK2206 and IF compared to that of either agent alone.
Together, the present study reports a novel use of IF in mitigating human colorectal cancer proliferation and inducing apoptosis via blockage of Akt pathway.
异秦皮啶（IF）是一种天然香豆素化合物，据报道在人类肝癌中具有抗癌活性。然而，IF是否参与结直肠癌的发生和发展调控尚未得到充分阐明。
分别通过细胞计数试剂盒-8（CCK-8）和集落形成试验评估细胞增殖。进行Transwell试验以评估细胞迁移和侵袭能力。此外，通过共聚焦显微镜分析、流式细胞术检测和TdT介导的dUTP缺口末端标记（TUNEL）法评估细胞凋亡。进行蛋白质印迹法检测相关蛋白的表达。
在此，结果表明IF以剂量和时间依赖性方式显著抑制人结肠癌细胞HT-29和SW-480的细胞增殖。此外，IF处理对HT-29和SW-480细胞的细胞集落形成显示出明显的抑制活性。共聚焦显微镜分析和流式细胞术检测显示，与对照相比，IF显著诱导HT-29和SW-480细胞凋亡。并且IF显著降低抗凋亡蛋白bcl-2的表达，而在HT-29和SW-480细胞中，包括caspase-3、caspase-9和bax在内的促凋亡蛋白的表达显著增加。此外，IF通过抑制p-Akt的表达阻断Akt通路。此外，Akt抑制剂MK2206可抑制细胞集落形成并诱导凋亡。与单独使用任何一种药物相比，在MK2206和IF共同存在时这种作用更为明显。
总之，本研究报道了IF通过阻断Akt通路减轻人结直肠癌增殖并诱导凋亡的新用途。
