Effect of Geniposide on Malignant Progression of Osteosarcoma Cells by Regulating the JAK2/STAT3/SOCS1 Signaling Pathway
LI Haixia1, MIAO Cunliang1 *, AN Zhihui1, LIU Guofeng1, YANG Donghai2
This study aims to investigate the effect of GEN (geniposide) on the malignant progression of osteosarcoma cells by regulating the JAK2 (janus kinase 2)/STAT3 (signal transducer and activator of transcription 3)/SOCS1 (suppressor of cytokine signaling 1) signaling pathway. Human osteosarcoma cells (U2OS) were treated with GEN at a concentration of 2.5-40 mg/mL, and cell activity was detected using the CCK-8 method to screen for the optimal drug concentration; U2OS cells were grouped into Control group, low, medium, and high concentration geniposide groups (GEN-L group, GEN-M group, GEN-H group), and high concentration geniposide+STAT3 activator group (GEN-H+colivelin group); cell proliferation was detected using colony formation assay; cell apoptosis was detected by flow cytometry; cell migration was detected using wound healing assay; cell invasion was detected using Transwell invasion assay; Western blot was applied to detect the expression of Cyclin D1, Ki67, Bax, Caspase-3, MMP-9, MMP-2, JAK2, STAT3, and SOCS1 proteins; nude mouse transplantation tumor experiment was applied to detect the effect of GEN on the growth of osteosarcoma transplantation tumors. 5 mg/mL, 10 mg/mL, and 20 mg/mL GEN were selected for the subsequent study. Compared with the Control group, the number of colony formation, scratch healing rate, number of cell invasion, and expression levels of Ki67, Cyclin D1, MMP-9, MMP-2, p-JAK2/JAK2, p-STAT3/STAT3 decreased in a concentration dependent manner in the GEN-L, GEN-M, and GEN-H groups, the expression of SOCS1 protein, cell apoptosis rate, and expression levels of Caspase-3 and Bax increased, in a concentration dependent manner (P<0.05); compared with the GEN-H group, the number of colony formation, scratch healing rate, number of cell invasion, and expression levels of Ki67, Cyclin D1, MMP-9, MMP-2, p-JAK2/JAK2, p-STAT3/STAT3 obviously increased in the GEN-H+colivelin group, the expression of SOCS1 protein, cell apoptosis rate, and expression levels of Caspase-3 and Bax obviously reduced (P<0.05). The transplantation experiment showed that the GEN group had slower growth of transplanted tumors compared to the Control group, the volume and mass of the transplanted tumor decreased, the expression levels of p-JAK2/JAK2, p-STAT3/STAT3 reduced, SOCS1 level increased (P<0.05). GEN can inhibit the malignant progression of osteosarcoma cells by regulating the JAK2/STAT3/SOCS1 signaling pathway.
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