GANT61 plays antitumor effects by inducing oxidative stress through the miRNA‐1286/RAB31 axis in osteosarcoma

Abstract

Osteosarcoma (OS) is a rare malignancy of bone associated with poor clinical outcomes. The antitumor effects of GANT61 on OS is unclear. To investigate antitumor effects and mechanism of GANT61 in OS cells and xenograft model. Effects of GANT61 on cell viability, clone formation, cell cycle, apoptosis, migration, and invasion ability of OS cells were assessed. Reactive oxygen species (ROS) levels measured by dichlorofluorescein fluorescence were used to evaluate oxidative stress. The Xenograft model was constructed to investigate the antitumor effects of GANT61 in vivo. The microRNA (miRNA)‐1286 was downregulated, while RAB31 upregulated in OS tissues and cells. GANT61 inhibited viability, migration, and invasion ability of OS cells (SaOS‐2 and U2OS), and induced apoptosis and the ROS production, along with miRNA‐1286 upregulation and RAB13 downregulation. After knockdown of miRNA‐1286, GANT6‐induced cell inhibition was attenuated, along with RAB31 upregulation. Inversely, miRNA‐1286 overexpression downregulated RAB31. Dual‐luciferase reporter assay verified that miR‐1286 negatively targeted RAB13. Moreover, the knockdown of RAB31 stimulated apoptosis and ROS production while inhibited viability, migration, and invasion of GANT61‐treated cells. In vivo experiments further confirmed that GANT61 inhibited tumor growth and RAB13 expression, but enhanced miRNA‐1286. The study demonstrated that GANT61 inhibited cell aggressive phenotype and tumor growth by inducing oxidative stress through the miRNA‐1286/RAB31 axis. Our findings provided a potential antitumor agent for the OS clinical treatment.

Silencing of circHIPK3 Sensitizes Paclitaxel-Resistant Breast Cancer Cells to Chemotherapy by Regulating HK2 Through Targeting miR-1286

“…31 Several previous reports had demonstrated the conflicting roles of miR-1286 in human carcinogenesis. 19,[32][33][34] These contradictory results might be in part due to the different types of tumors in these reports, where miR-1286 exerted an anti-tumor property in osteosarcoma 19,32 and contributed to the progression of cutaneous melanoma 33 and lung adenocarcinoma. 34 Importantly, miR-1286 expression was remarkably altered by cisplatin or 5-fluorouracil chemotherapy in esophageal cancer cells.…”

Evaluation of Hedgehog Pathway Inhibitors as a Therapeutic Option for Uterine Leiomyosarcoma Using the Xenograft Model

“…In thyroid tumors, the treatment downregulated the GLI1 protein expression and reduced the tumor volume [ 48 ]. In a xenograft model of osteosarcoma, Gant61 promoted tumor regression ( 49 ). c-MYC and BMP4 have been described as GLI target genes [ 58 , 59 ].…”

“…After the tumor development, the animals were randomized separately into three groups, SMO inhibitor LDE225 ( n = 5), GLI inhibitor Gant61 ( n = 6 × 2), and control ( n = 6 × 2). Gli treatments were repeated compared to the vehicle control group to receive enough tissues for cellular and molecular analysis due to Gant61’s marked inhibition of tumor growth; 20 mg/kg of LDE225 [ 47 ], 20 mg/kg of Gant61 [ 20 , 48 , 49 ], or corn oil (vehicle) were administrated via oral gavage three times per week for 10 days to the Gant61 group and 21 days to the LDE225 group. After the treatment, the animals were sacrificed, and tumors were collected for histopathological and RNA and protein expression profile analysis.…”

Sonic Hedgehog Signature in Pediatric Primary Bone Tumors: Effects of the GLI Antagonist GANT61 on Ewing’s Sarcoma Tumor Growth

“…For example, Hirotsu et al showed that cyclopamine, an SMO inhibitor, and a shRNA against SMO both inhibit the proliferation of 143B and HOS OS cells and the growth of the primary tumor [ 34 ]. One recent study demonstrated that the Gli1/2 inhibitor GANT61 is able to inhibit the growth of OS tumors by inducing oxidative stress via the miRNA-1286/RAB31 axis using an OS xenograft model [ 35 ].…”