乙酰哈巴苷通过Wnt信号通路诱导结肠癌HCT116细胞凋亡

8-O-Acetylharpagide induces apoptosis of colon cancer HCT116 cells through Wnt signaling pathway

本文探讨乙酰哈巴苷诱导结肠癌HCT116细胞凋亡机制。采用流式细胞术检测乙酰哈巴苷对HCT116细胞周期及凋亡的影响,发现细胞周期被阻滞在G1期,0.75 mmol/L乙酰哈巴苷处理48 h后,细胞凋亡明显增加。转录组测序显示乙酰哈巴苷引起HCT116细胞1921个mRNA上调和2208个mRNA下调,可显著影响Wnt信号通路。RT-qPCR和Western blot验证实验表明,乙酰哈巴苷处理后,HCT116细胞中β-catenin、CyclinD1、Survivin、Bcl-2和c-Myc蛋白显著降低(P<0.05),凋亡相关蛋白Bax和cleaved-caspase3表达增加(P<0.05)。上述结果表明,乙酰哈巴苷主要通过Wnt信号通路抑制结肠癌细胞增殖并诱导细胞凋亡。

In this study,we investigated the effect and molecular mechanism of 8-O-acetylharpagide in colon cancer.Human colon cancer line HCT116 cell cycle was blocked in G1 phase after 8-O-acetylharpagide treatment.In addition,0.75 mmol/L 8-O-acetylharpagide significantly increased apoptosis of HCT116 cells.The whole transcriptome sequencing analysis showed that 8-O-acetylharpagide caused a total of 1921 up-regulated mRNAs and 2208 down-regulated mRNAs in HCT116 cells,and Wnt signaling pathway-related genes transcription level changed significantly.In cellular experiments in vitro,8-O-acetylharpagide treatment led to significant down-regulation ofβ-catenin,CyclinD1,Survivin,c-Myc and Bcl-2,while significant up-regulation of proapoptotic proteins Bax and cleaved-caspase3.Together,these results suggest that 8-O-acetylharpagide inhibit the proliferation and induce apoptosis of colon cancer cells mainly through Wnt signaling pathway.