Low-frequency ultrasound enhances chemotherapy sensitivity and induces autophagy in PTX-resistant PC-3 cells via the endoplasmic reticulum stress-mediated PI3K/Akt/mTOR signaling pathway

• Main Authors: Wu Y, Liu X, Qin Z, Hu L, Wang X
• Format: Article
• Language: English
• Published: Dove Medical Press 2018-09-01
• Series: OncoTargets and Therapy
• Subjects: Autophagy; Multidrug resistance; Sonodynamic therapy; Endoplasmic reticulum stress; apoptosis
• Online Access: https://www.dovepress.com/low-frequency-ultrasound-enhances-chemotherapy-sensitivity-and-induces-peer-reviewed-article-OTT

Yuqi Wu,1 Xiaobing Liu,2 Zizhen Qin,1 Li Hu,1 Xiangwei Wang1 1Department of Urology, Carson International Cancer Center, Shenzhen University General Hospital & Shenzhen University Clinical Medical Academy Center, Shenzhen University, Shenzhen, People’s Republic of China; 2Department of Urology, Second Affiliated Hospital, Third Military Medical University, Chongqing, People’s Republic of China Background: Sonodynamic therapy (SDT) is an emerging tumor-inhibiting method that has gained attention in cancer therapy in the last several years. Although autophagy has been observed in SDT-treated cancer cells, its role and mechanism of action remain unclear. This study aimed to investigate the effects of low-frequency ultrasound on autophagy and drug-resistance of paclitaxel (PTX)-resistant PC-3 cells via the endoplasmic reticulum stress (ERs)-mediated PI3K/AT/mTOR signaling pathway. Methods: CCK-8 assay was conducted to select the appropriate exposure time for PTX-resistant PC-3 cells under low-frequency ultrasound. PTX-resistant PC-3 cells were divided into a control group, PTX group, ultrasound group, ultrasound + PTX group, ultrasound + PTX + autophagy-related gene 5 (Atg5) siRNA group, and ultrasound + 4-PBA (an ERs inhibitor) group. Autophagy was observed by transmission electron microscopy (TEM) and fluorescence microscopy. Cell proliferation was evaluated using CCK-8 assay; apoptosis was detected by flow cytometry. Expression of multiple drug-resistance genes was detected by qRT-PCR. Western blotting was used to detect the expression of ERS-related proteins, autophagy-related proteins, apoptosis-related proteins, and PI3K/AKT/mTOR pathway-related proteins. Results: Ten-second exposure was selected as optimal for all experiments. Compared to the PTX group, the level of autophagy, inhibition rate, apoptosis rate, and expression of ERS-related proteins (GRP78) increased, whereas the expression of multiple drug-resistance genes (MRP3, MRP7, and P-glycoprotein), PI3K/AKT/mTOR pathway-related proteins (PI3K, p-AKT, mTORC1), and apoptosis-related proteins (Bcl-2, NF-κB) decreased in PTX-resistant PC-3 cells after low-frequency ultrasound and PTX treatment for 24 h. These trends were more obvious after treatment with Atg5 siRNA, excluding the autophagy level. Post 4-PBA-treatment, the expression of GRP78 and LC3II proteins decreased, whereas that of PI3K, p-AKT, and mTORC1 increased. Conclusion: Results indicated that ultrasound induces autophagy by ERs-mediated PI3K/AKT/mTOR signaling pathway in PTX-resistant PC-3 cells; this autophagy acts as a cytoprotector during low-frequency ultrasound-mediated reversal of drug resistance. Keywords: prostate cancer, multidrug resistance, sonodynamic therapy, autophagy, apoptosis, endoplasmic reticulum stress