Won Shen-Jeu, Yen Cheng-Hsin, Lin Ting-Yu, Jiang-Shieh Ya-Fen, Lin Chun-Nan, Chen Jyun-Ti, Su Chun-Li
Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, Tainan, Taiwan.
Department of Human Development and Family Studies, National Taiwan Normal University, Taipei, Taiwan.
J Cell Physiol. 2018 Jan;233(1):497-505. doi: 10.1002/jcp.25910. Epub 2017 May 3.
The tautomeric pair of garcinielliptone FC (GFC) is a novel tautomeric pair of polyprenyl benzophenonoid isolated from the pericarps of Garcinia subelliptica Merr. (G. subelliptica, Clusiaceae), a tree with abundant sources of polyphenols. Our previous report demonstrated that GFC induced apoptosis on various types of human cancer cell lines including chemoresistant human colorectal cancer HT-29 cells. In the present study, we observed that many autophagy-related genes in GFC-treated HT-29 cells were up- and down-regulated using a cDNA microarray containing oncogenes and kinase genes. GFC-induced autophagy of HT-29 cells was confirmed by observing the formation of acidic vesicular organelles, LC3 puncta, and double-membrane autophagic vesicles using flow cytometry, confocal microscopy, and transmission electron microscopy, respectively. Inhibition of AKT/mTOR/P70S6K signaling as well as formation of Atg5-Atg12 and PI3K/Beclin-1 complexes were observed using Western blot. Administration of autophagy inhibitor (3-methyladenine and shRNA Atg5) and apoptosis inhibitor Z-VAD showed that the GFC-induced autophagy was cytotoxic form and GFC-induced apoptosis enhanced GFC-induced autophagy. Our data suggest the involvement of autophagy and apoptosis in GFC-induced anticancer mechanisms of human colorectal cancer.
藤黄椭圆酮FC（GFC）的互变异构对是从多花山竹子（藤黄科）的果皮中分离出的一种新型的聚异戊二烯基二苯甲酮互变异构对，该植物富含多酚。我们之前的报告表明，GFC可诱导多种类型的人类癌细胞系发生凋亡，包括对化疗耐药的人类结肠直肠癌HT-29细胞。在本研究中，我们使用包含癌基因和激酶基因的cDNA微阵列观察到，GFC处理的HT-29细胞中许多自噬相关基因的表达上调和下调。分别使用流式细胞术、共聚焦显微镜和透射电子显微镜观察酸性囊泡细胞器、LC3斑点和双膜自噬泡的形成，证实了GFC诱导HT-29细胞发生自噬。使用蛋白质免疫印迹法观察到AKT/mTOR/P70S6K信号通路的抑制以及Atg5-Atg12和PI3K/Beclin-1复合物的形成。给予自噬抑制剂（3-甲基腺嘌呤和Atg5短发夹RNA）和凋亡抑制剂Z-VAD表明，GFC诱导的自噬是细胞毒性形式，且GFC诱导的凋亡增强了GFC诱导的自噬。我们的数据表明自噬和凋亡参与了GFC诱导的人类结肠直肠癌抗癌机制。
