Abstract
Osteosarcoma (OS) is the most common malignancy of bone. Liensinine exerts antitumor effects on cancers of the colon, breast, and gallbladder. However, its antitumor activity in OS remains unclear. This study is aimed at investigating the efficacy of liensinine against OS and the underlying mechanism of action. Cell proliferation, apoptosis, and cycle arrest in OS were detected using the Cell Counting Kit-8 (CCK-8), colony formation, and flow cytometry assays, respectively. The production of reactive oxygen species (ROS), glutathione (GSH) and glutathione disulfide (GSSG) concentrations, and mitochondrial membrane potential (MMP) of OS cells were measured by flow cytometry, colorimetry, and JC-1 staining. The expressions of factors related to apoptosis, cell cycle, and activation of the JAK2/STAT3 pathway were determined by Western blotting. To examine the potential role of ROS, an antioxidant (N-acetyl cysteine, NAC) was used in combination with liensinine. In vivo, we generated a xenograft mouse model to assess its antitumor efficacy. Tissue level expressions of factors related to apoptosis and activation of the JAK2/STAT3 pathway were assessed by immunohistochemistry or Western blotting. Liensinine inhibited the proliferation and induced G0/G1 phase arrest and apoptosis of OS cells in a dose-dependent manner. Additionally, liensinine promoted intracellular ROS production, enhanced the GSSG/GSH ratio, and induced MMP loss and ROS-mediated suppression of the JAK2/STAT3 pathway. NAC significantly attenuated the liensinine-induced antitumor activities and activated the JAK2/STAT3 pathway. In vivo, liensinine effectively inhibited the OS growth and promoted apoptosis; however, it had no negative effect on the internal organs. In conclusion, liensinine-induced ROS production could suppress the activation of the JAK2/STAT3 pathway and inhibit the OS growth both in vivo and in vitro. Our findings provided a new rationale for subsequent academic and clinical research on OS treatment.
Highlights
Osteosarcoma (OS) is a malignant tumor originating from mesenchymal tissues, accounts for 20% of the primary malignant bone acanthomas; it is the most common primary malignant bone tumor in adolescents [1]
Cell viability was evaluated by the Cell Counting Kit-8 (CCK-8) assay, which showed that liensinine at 5 μM, 10 μM, 20 μM, 40 μM, and 80 μM did not inhibit the viability of hFOB 1.19 cells (Figure 1(b)); it significantly reduced the viability of SaOS-2 (Figure 1(c)), MG-63 (Figure 1(d)), 143B (Figure 1(e)), and U2OS (Figure 1(f)) cells
We evaluated the effect of the same concentration of liensinine on apoptosis and JAK2/STAT3 pathway activation (p-STAT3, STAT3, p-JAK2, and JAK2) of hFOB 1.19 cells
Summary
Osteosarcoma (OS) is a malignant tumor originating from mesenchymal tissues, accounts for 20% of the primary malignant bone acanthomas; it is the most common primary malignant bone tumor in adolescents [1]. OS is characterized by high malignancy, rapid growth, early metastasis, and poor prognosis. It is the leading cause of cancerrelated death among children and adolescents [2, 3]. Surgery combined with multiagent chemotherapy is the standard treatment regime for OS [2, 4]. Longterm chemotherapy causes many irreversible systemic side effects, including cardiotoxicity, secondary malignancies, neurotoxicity, and infertility. Physical disability caused by surgical procedures has a great impact on the mental health and quality of life of the patients [2, 5]. The development of novel anti-OS drugs is important for effectively inhibiting tumor progression and prolonging the survival duration of patients
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