Diallyl trisulfide regulates cell apoptosis and invasion in human osteosarcoma U2OS cells through regulating PI3K/AKT/GSK3β signaling pathway

Pan He¹, Zhijun Wang¹, Bin Sheng¹, Yongqiang Xu¹, Siyin Feng¹, Yan Huang¹, Fuqiang Gong², Liting Tang² and Liming Xie<sup>2

1</sup>The Department of Traumatic and Osteopathology, Hunan Provincial People's Hospital, Changsha and ²The First Affiliated Hospital, University of South China, Hengyang, Hunan, China

Offprint requests to: Pan He, The Department of Traumatic and Osteopathology, Hunan provincial people's hospital, Changsha, Hunan, China. e-mail: h13487490749@126.com and Liming Xie, The First Affiliated Hospital, University of South China, 69 Chuanshan Road, Hengyang, 421001, China. e-mail: xlmusc@163.com

Summary. Aims. To investigate the effects and the mechanisms of action of Diallyl trisulfide (DATS) on the proliferation and metastasis of human osteosarcoma (OS) U2OS. Methods. U2OS cells were treated by different concentrations of DATS at different time points. Cell proliferations were measured by MTT assay. DATS induced cell cycle distribution and apoptosis were evaluated by flow cytometry (FCM) with Annexin-V. Cell migration and invasion were detected by wound healing assay and transwell assay. The effects of DATS in U2OS cell growth and metastasis were also detected in a mouse OS xenograft model. Results. A time- and concentration-dependent cytotoxic effect of DATS was observed in U2OS cells. FCM with PI staining and Annexin-V -FITC indicated that DATS induces apoptosis and a G0/G1 cell cycle arrest of U2OS cells at all concentrations from 25 μmol/l to 100 μmol/l. DATS also inhibits the migration and invasion of U2OS cells. Western blot showed that the expression levels of p-AKT, p-GSK3β, Bcl-2, Vimentin and β-catenin were decreased, while the expression levels of Bad, Bax and E-cadherin were significantly increased in DATS treated U2OS cells. Analysis using a mouse xenograft model indicated that xenografts of DATS treatment group had a significant decrease in tumor volume and weight compared to the control group. Lung metastasis models in mice demonstrated that treatment of DATS inhibits lung metastasis of OS in vivo. Conclusions. These data suggested that DATS inhibits OS development and progression through the regulation of PI3K/AKT/GSK3β signaling pathways, accompanied by downregulation of Bcl-2, Vimentin and β-catenin, as well as upregulation of Bad, Bax and E-cadherin. Therefore, our data demonstrated that DATS exerted its anticancer effects by inhibiting cell proliferation, migration and invasion in vitro and in vivo. These results provide evidence for the use of the natural product DATS either alone or in combination with standard therapy for OS. Histol Histopathol 35, 1511-1520 (2020)

Key words: Osteosarcoma, Diallyl trisulfide, p-AKT, p-GSK3β

DOI: 10.14670/HH-18-299