Xu Yuehua, Cai Xueting, Zong Bin, Feng Rui, Ji Yali, Chen Gang, Li Zhongxing
Zhenjiang Hospital of Chinese Traditional and Western Medicine, Zhenjiang, China.
Affiliated Hospital of Integrated Traditional Chinese and Western Medicine, Nanjing University of Chinese Medicine, Nanjing, China.
Front Pharmacol. 2018 Apr 4;9:234. doi: 10.3389/fphar.2018.00234. eCollection 2018.
Qianlie Xiaozheng decoction (QLXZD), a traditional Chinese medicinal formula, has been used clinically to treat advanced prostate cancer (PCa) for more than 10 years. However, experimental evidence supporting its efficacy is lacking. Here, we investigated the anticancer properties and molecular mechanism of QLXZD in a human PCa cell line (PC3) and using PC3 xenografts in nude mice. We confirmed the antineoplastic activity of QLXZD by analyzing cell viability and tumor volume growth, which decreased significantly compared to that of the controls. Autophagy following QLXZD treatment was detected morphologically using transmission electron microscopy and was confirmed by measuring the expression of autophagy markers (LC3-II and p62) using fluorescence analysis, flow cytometry, and western blotting. Increasing autophagic flux induced by QLXZD was monitored via pmCherry-GFP-LC3 fluorescence analysis. QLXZD-induced autophagic cell death was alleviated by the autophagy inhibitors, 3-methyl adenine and hydroxychloroquine. We evaluated the total expression and phosphorylation levels of proteins involved in the Akt/mTOR pathway regulating autophagy. Phosphorylation of Akt, mTOR, and p70S6K, but not total protein levels, decreased following treatment. This is the first study to demonstrate the autophagy-related mechanistic pathways utilized during QLXZD-mediated antitumor activity both and . These findings support the clinical use of QLXZD for PCa treatment.
前列消症汤（QLXZD）是一种中药方剂，已在临床上用于治疗晚期前列腺癌（PCa）超过10年。然而，缺乏支持其疗效的实验证据。在此，我们在人PCa细胞系（PC3）中以及使用PC3裸鼠异种移植模型研究了QLXZD的抗癌特性和分子机制。我们通过分析细胞活力和肿瘤体积增长来确认QLXZD的抗肿瘤活性，与对照组相比，其显著降低。使用透射电子显微镜从形态学上检测QLXZD处理后的自噬，并通过荧光分析、流式细胞术和蛋白质印迹法测量自噬标志物（LC3-II和p62）的表达来进行确认。通过pmCherry-GFP-LC3荧光分析监测QLXZD诱导的自噬通量增加。自噬抑制剂3-甲基腺嘌呤和羟氯喹减轻了QLXZD诱导的自噬性细胞死亡。我们评估了参与调节自噬的Akt/mTOR途径中蛋白质的总表达和磷酸化水平。处理后，Akt、mTOR和p70S6K的磷酸化水平降低，但蛋白质总水平未降低。这是第一项证明QLXZD介导的抗肿瘤活性过程中所利用的自噬相关机制途径的研究。这些发现支持QLXZD在PCa治疗中的临床应用。
