Lnc-CLSTN2-1:1 Promotes Osteosarcoma Progression by Disrupting Redox Balance through PI3K/AKT Signaling Pathway

Hao Lin; Xinjian Wei; Junhong Ye; Jiaxian Chen; Jing Huang; Tingrui Wu; Zhenju Chen; Yuming Zeng; Lijiao Peng

doi:10.7150/jca.91579

Lnc-CLSTN2-1:1 Promotes Osteosarcoma Progression by Disrupting Redox Balance through PI3K/AKT Signaling Pathway

J Cancer. 2024 Jan 12;15(5):1287-1298. doi: 10.7150/jca.91579. eCollection 2024.

Authors

Hao Lin¹, Xinjian Wei¹, Junhong Ye¹, Jiaxian Chen¹, Jing Huang², Tingrui Wu¹, Zhenju Chen³, Yuming Zeng¹, Lijiao Peng²

Affiliations

¹ Department of Orthopedics, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong,534001, China.
² Oncology Hospital, Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong,534001, China.
³ Department of Orthopedics, Suixi Hospital of Affiliated Hospital of Guangdong Medical University, Zhanjiang, Guangdong,534001, China.

Abstract

Objective: Most patients with osteosarcoma (OS) have an extremely poor prognosis. The primary purpose of this investigation was to explore the biological effect of Lnc-CLSTN2-1:1 on OS and the potential processes involved. Materials and procedures: We selected differentially overexpressed Lnc-CLSTN2-1:1 from our laboratory's existing RNA sequence analysis data (fibroblast osteoblast (hFOB 1.19) and three osteosarcoma cell lines (HOS, MG63, and U2OS) as the research object. Next, we detected Lnc-CLSTN2-1:1 in the osteosarcoma HOS cell line and fibroblast cells using qRT-PCR. We evaluated cell proliferation ability using EdU incorporation test, CCK-8 test, and cell clone formation; cell invasion and migration were assessed using the Transwell test, while flow cytometry examined cell cycle, apoptosis, and reactive oxygen species (ROS); Subsequently, the activity changes of selenase (GPx) glutathione peroxidase and (TrxR) thioredoxin reductase were detected. In addition, changes in related proteins were analyzed through Western blotting. Results: The expression of Lnc-CLSTN2-1:1 in osteosarcoma cells was significantly increased. The proliferation, invasion, and migration of osteosarcoma cells were significantly inhibited by knockdown of the expression of Lnc-CLSTN2-1:1, and the cell cycle-related signaling pathway PI3K/AKT/GSK-3β/cycinD1 was also inhibited. However, insulin-like growth factor-1 (igf-1) could reverse this process. In addition, we examined the activity of two selenophenases (TrxR and GPx) and the changes of ROS before and after Lnc-CLSTN2-1:1 knockdown. The results showed that both TrxR and GPx activities were reduced after Lnc-CLSTN2-1:1 knockdown, resulting in the inhibition of antioxidant stress levels, while intracellular ROS levels were high, which eventually caused killing effects on tumor cells due to the imbalance between oxidative stress and antioxidant stress. Conclusion: Our results showed that Lnc-CLSTN2-1:1 enhanced anti-oxidative stress TrxR and GPx selenoprotein activities through the PI3K/AKT signaling pathway while counteracting the loss of reactive oxygen species ROS produced by mitochondria to osteosarcoma cells, which protected osteosarcoma cells and thus promoted the proliferation and metastatic ability of OS.

Keywords: GPx; ROS; TrxR; long non-coding RNA; osteosarcoma.