Qianlie Xiaozheng Decoction Induces Autophagy in Human Prostate Cancer Cells via Inhibition of the Akt/mTOR Pathway

Abstract

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 vitro in a human PCa cell line (PC3) and in vivo 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 in vitro and in vivo. These findings support the clinical use of QLXZD for PCa treatment.

Autophagy-related Proteins as a Prognostic Factor of Patients With Colorectal Cancer

“…Several clinical trials on patients with metastatic colorectal cancer have shown that chloroquine and HCQ are useful only when they are combined with chemotherapeutic agents, inhibitors of histone deacetylases and antiangiogenic agents 45,46. Inhibition of autophagy with molecules such as HCQ and 3-MA is identified by the accumulation of p62 and LC3B in CRC cell lines 47,48. Furthermore, 3-MA inhibits the first steps of autophagy and seems to reduce the protein levels of Beclin-1 in 3 out of 4 used MSS CRC cell lines.…”

Timosaponin�A‑III induces autophagy of T‑cell acute lymphoblastic leukemia Jurkat cells via inhibition of the PI3K/Akt/mTOR pathway

“…The results demonstrated that TAIII induces the expression of the autophagy-associated proteins Beclin 1 and LC3-II, while reducing the proliferation of Jurkat cells in vitro. There have been a substantial amount of studies indicating that PI3K/Akt/mTOR signaling are key axes involved in anticancer drug-induced autophagy (8,34,35). Further investigation was performed, with a conclusion that the PI3K/Akt/mTOR signaling pathway was inactivated by TAIII through suppressing the expression of p-PI3K, p-Akt and p-mTOR.…”

Unravelling the Anticancer Mechanisms of Traditional Herbal Medicines with Metabolomics

“…Qianlie Xiaozheng decoction (QLXZD), a polyherbal formulation containing Coix lacryma-jobi and six other herbs, is clinically used to treat prostate cancer. The only mechanistic study on QLXZD to date reported its autophagyrelated toxicity to involve inhibition of the Akt/mTOR pathway [146] in PC3 cells and PC3 xenografts mice.…”