New benzimidazole acridine derivative induces human colon cancer cell apoptosis in vitro via the ROS-JNK signaling pathway  

作　　者：Kang CHEN Bi-zhu CHU Feng LIU Bin LI Chun-mei GAO Lu-lu LI Qin-sheng SUN Zhi-fa SHEN Yu-yang JIANG 

机构地区：[1]Department of Chemistry, Tsinghua University, Beijing 100084, China [2]The Ministry-Province Jointly Constructed Base for State Key Lab-Shenzhen Key Laboratory of Chemical Biology, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China [3]Shenzhen Anti-Tumor Drug Development Engineering Laboratory, Graduate School at Shenzhen, Tsinghua University, Shenzhen 518055, China [4]Shenzhen Kivita Innovative Drug Discovery Institute, Shenzhen 518055, China [5]School of Medicine, Tsinghua University, Beijing 100084, China

出　　处：《Acta Pharmacologica Sinica》2015年第9期1074-1084,共11页中国药理学报（英文版）

基　　金：This work was supported by grants from the National Natural Science Foundation of China （21272134 and 21402105） and the Shenzhen Municipal government SZSITIC （ZDSY20120619141412872, JCYJ20130402164027386, KC2013ZDZJ0019A and CXB201104210013A）.

摘　　要：Aim： To investigate the mechanisms underlying anticancer action of the benzimidazole acridine derivative N-{（1H-benzo[d]imidazol-2-yl） methyl}-2-butylacridin-9-amine （8m） against human colon cancer cells in vitro. Methods： Human colon cancer cell lines SW480 and HCT116 were incubated in the presence of 8m, and then the cell proliferation and apoptosis were measured. The expression of apoptotic/signaling genes and proteins was detected using RT-PCR and Western blotting. ROS generation and mitochondrial membrane depolarization were visualized with fluorescence microscopy. Results： 8m dose-dependently suppressed the proliferation of SW480 and HCT116 cells with ICso values of 6.77 and 3.33 pmol/L, respectively. 8m induced apoptosis of HCT116 cells, accompanied by down-regulation of Bcl-2, up-regulation of death receptor-5 （DR5）, truncation of Bid, cleavage of PARP, and activation of caspases （including caspase-8 and caspase-9 as well as the downstream caspases-3 and caspase-7）. Moreover, 8m selectively activated JNK and p38 without affecting ERK in HCT116 cells. Knockout of JNK1, but not p38, attenuated 8m-induced apoptosis. In addition, 8m induced ROS production and mitochondrial membrane depolarization in HCT116 cells. Pretreatment with the antioxidants N-acetyl cysteine or glutathione attenuated 8m-induced apoptosis and JNK activation in HCT116 cells. Conclusion： The new benzimidazole acridine derivative, 8m exerts anticancer activity against human colon cancer cells in vitro by inducing both intrinsic and extrinsic apoptosis pathways via the ROS-JNK1 pathway.Aim： To investigate the mechanisms underlying anticancer action of the benzimidazole acridine derivative N-{（1H-benzo[d]imidazol-2-yl） methyl}-2-butylacridin-9-amine （8m） against human colon cancer cells in vitro. Methods： Human colon cancer cell lines SW480 and HCT116 were incubated in the presence of 8m, and then the cell proliferation and apoptosis were measured. The expression of apoptotic/signaling genes and proteins was detected using RT-PCR and Western blotting. ROS generation and mitochondrial membrane depolarization were visualized with fluorescence microscopy. Results： 8m dose-dependently suppressed the proliferation of SW480 and HCT116 cells with ICso values of 6.77 and 3.33 pmol/L, respectively. 8m induced apoptosis of HCT116 cells, accompanied by down-regulation of Bcl-2, up-regulation of death receptor-5 （DR5）, truncation of Bid, cleavage of PARP, and activation of caspases （including caspase-8 and caspase-9 as well as the downstream caspases-3 and caspase-7）. Moreover, 8m selectively activated JNK and p38 without affecting ERK in HCT116 cells. Knockout of JNK1, but not p38, attenuated 8m-induced apoptosis. In addition, 8m induced ROS production and mitochondrial membrane depolarization in HCT116 cells. Pretreatment with the antioxidants N-acetyl cysteine or glutathione attenuated 8m-induced apoptosis and JNK activation in HCT116 cells. Conclusion： The new benzimidazole acridine derivative, 8m exerts anticancer activity against human colon cancer cells in vitro by inducing both intrinsic and extrinsic apoptosis pathways via the ROS-JNK1 pathway.

关 键 词：benzimidazole acridine anticancer drug colon cancer apoptosis death receptor-5 JNK1 ROS 

分 类 号：Q255[生物学—细胞生物学] Q813