Liensinine Inhibits Osteosarcoma Growth by ROS-Mediated Suppression of the JAK2/STAT3 Signaling Pathway

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.

KLF5‑induced miR‑487a augments the progression of osteosarcoma cells by targeting NKX3‑1 in vitro

“…It was hypothesized that ROS may also serve a crucial role in development of osteosarcoma. Jia et al revealed that liensinine inhibits osteosarcoma growth by ROS-mediated suppression of the JAK2/STAT3 signaling pathway ( 35 ) and Rawat and Nayak ( 36 ) reported that Piperlongumine induces ROS-mediated apoptosis by transcriptional regulation of SMAD4/P21/P53 genes and synergizes with doxorubicin in osteosarcoma cells.…”

Liensinine inhibits progression of intrahepatic cholangiocarcinoma by regulating TGF‐β1 /P‐smad3 signaling through HIF‐1a

“…Previous studies have shown that liensinine may be associated with the induction of tumor cell apoptosis. 22,24 Therefore, we examined apoptosis in ICC cells following liensinine treatment. However, we found that liensinine did not significantly induce apoptosis in ICC cells, and apoptosisrelated proteins showed no significant trend in Western blot analysis (Figures S1C,D).…”

“…20 In 2015, it was the first time that liensinine was identified as an autophagy/mitophagy inhibitor that could inhibit the growth of breast cancer. 21 Besides, this alkaloid has also been demonstrated to inhibit growth of osteosarcoma cells, 22 gallbladder, and colon cancer cells, 23,24 The secondary antibodies were purchased from BioSharp. HIF-1a overexpressed plasmid was from Genechem.…”

ROS-Generating Hyaluronic Acid-Modified Zirconium Dioxide-Acetylacetonate Nanoparticles as a Theranostic Platform for the Treatment of Osteosarcoma

“…This material shows intriguing photoluminescent properties in the visible range [ 21 ], which, combined with the capability of producing ROS, make the hybrid ZrO 2 a powerful theranostic material for cancer therapy. In particular, the generation of ROS has been used as a synergistic process in the treatment of osteosarcoma (OS) to increase the efficacy of chemotherapeutic drugs [ 22 ]. OS is a bone cancer occurring most often in children and young adults between the age of 10 and 20 years [ 23 ].…”

“…Numerous studies have reported the overexpression of surface molecules on the OS cells, including CD44, the major hyaluronic acid (HA) receptor [ 27 ]. The binding between CD44 and HA has been proven to be involved in multidrug resistance [ 28 ], tumor progression, metastasis, and proliferation [ 22 ]. Numerous ROS-generating NPs targeting CD44 + cells have been proposed [ 23 , 29 , 30 ], but, to the best of our knowledge, none of them combines imaging and therapy in a label-free and light-independent approach, such as our ZrO 2 -acac NPs.…”