Liao Kuan-Fu, Chiu Tsung-Lang, Huang Sung-Ying, Hsieh Teng-Fu, Chang Shu-Fang, Ruan Jhen-Wei, Chen Shee-Ping, Pang Cheng-Yoong, Chiu Sheng-Chun
Graduate Institute of Integrated Medicine, China Medical University, Taichung, Taiwan.
Department of Internal Medicine, Taichung Tzu Chi Hospital, Buddhist Tzu Chi Medical Foundation, Taichung, Taiwan.
Cell Physiol Biochem. 2018;48(6):2231-2246. doi: 10.1159/000492641. Epub 2018 Aug 16.
BACKGROUND/AIMS: Radix Angelica Sinensis (danggui in Chinese) is widely used in traditional chinese medicine (TCM). N-butylidenephthalide (BP), a bioactive compound in danggui, is a potential antitumor agent for various cancer types. However, its clinical effect and mechanism in the treatment of gastric cancer remain undetermined.
The in vivo protective effect of danggui in patients with gastric cancer were validated using data from Taiwan's National Health Insurance Research Database (NHIRD). The genes induced by BP-treatment were analyzed by whole transcriptome RNA sequencing (RNA-seq) and validated by real-time PCR, western blot and siRNA transfection. The effect of BP on AGS cell migration and invasion was evaluated in transwell assays. The antitumor effects of BP were evaluated in vivo in an AGS xenograft animal model.
Danggui users were found to have an increased survival rate when compared with danggui nonusers (log-rank test p = 0.002) . The use of danggui highly associated with decreased mortality (the adjusted hazard ratio (HR) of danggui user was 0.72 [95 % CI, 0.57-0.92] (p = 0.009). The in vitro results showed that BP inhibited gastric cancer cell proliferation, and triggered cellular apoptosis depending on the activation of mitochondrial apoptotic pathway. Using RNA-seq analysis we found that REDD1 was the highest transcript induced by BP in gastric cancer cells. BP induce an increase of REDD1 expression that inhibits mTOR signaling, thus inhibiting gastric cancer growth. We used RNA interference to demonstrate that the knock-down of REDD1 attenuated the BP-induced mTORC1 activation and growth inhibition. BP suppressed the growth of AGS xenografts tumor in vivo.
Danggui can prolong the survival rate of gastric cancer patients in Taiwan. BP caused gastric cancer cell death through the activation of mitochondria-intrinsic pathway and induced the REDD1 expression leading to mTOR signal pathway inhibition in gastric cancer cells. BP inhibited the in vivo growth of AGS xenograft tumors. These results may provide the basis for a new therapeutic approach toward the treatment of gastric cancer progression.
背景/目的：当归在中国传统医学中被广泛应用。正丁烯基苯酞（BP）是当归中的一种生物活性化合物，是多种癌症类型的潜在抗肿瘤药物。然而，其在胃癌治疗中的临床效果和机制仍未明确。
利用台湾国民健康保险研究数据库（NHIRD）的数据验证当归对胃癌患者的体内保护作用。通过全转录组RNA测序（RNA-seq）分析BP处理诱导的基因，并通过实时PCR、蛋白质印迹和小干扰RNA转染进行验证。在Transwell实验中评估BP对AGS细胞迁移和侵袭的影响。在AGS异种移植动物模型中体内评估BP的抗肿瘤作用。
与未使用当归的患者相比，使用当归的患者生存率有所提高（对数秩检验p = 0.002）。当归的使用与死亡率降低高度相关（当归使用者的调整后风险比（HR）为0.72 [95%置信区间，0.57 - 0.92]（p = 0.009））。体外结果表明，BP抑制胃癌细胞增殖，并通过激活线粒体凋亡途径引发细胞凋亡。通过RNA-seq分析，我们发现REDD1是BP在胃癌细胞中诱导表达最高的转录本。BP诱导REDD1表达增加，抑制mTOR信号传导，从而抑制胃癌生长。我们使用RNA干扰证明，敲低REDD1可减弱BP诱导的mTORC1激活和生长抑制。BP在体内抑制AGS异种移植肿瘤的生长。
当归可延长台湾地区胃癌患者的生存率。BP通过激活线粒体内在途径导致胃癌细胞死亡，并诱导REDD1表达，从而抑制胃癌细胞中的mTOR信号通路。BP抑制AGS异种移植肿瘤的体内生长。这些结果可能为胃癌进展的新治疗方法提供依据。
