HO-1 Regulates the Apoptosis of Osteosarcoma Cells through the P62/RelA Signaling Axis

作者全名："Chen, Lin; Zou, Luetao; Lei, Miao; Zhang, Xiaojun; Jiang, Wei; Hu, Zhenming"

作者地址："[Chen, Lin; Lei, Miao; Zhang, Xiaojun; Jiang, Wei; Hu, Zhenming] Chongqing Med Univ, Affiliated Hosp 1, Dept Orthoped, Chongqing 400016, Peoples R China; [Chen, Lin] Chongqing Univ Three Gorges Hosp, Dept Orthoped, Chongqing 404000, Peoples R China; [Zou, Luetao] Chongqing Univ Cent Hosp, Dept Orthoped, Chongqing 400015, Peoples R China; [Hu, Zhenming] Chongqing Med Univ, Univ Town Hosp, Dept Orthoped, Chongqing 401331, Peoples R China"

通信作者："Jiang, W; Hu, ZM (通讯作者)，Chongqing Med Univ, Affiliated Hosp 1, Dept Orthoped, Chongqing 400016, Peoples R China.; Hu, ZM (通讯作者)，Chongqing Med Univ, Univ Town Hosp, Dept Orthoped, Chongqing 401331, Peoples R China."

来源：JOURNAL OF BIOLOGICAL REGULATORS AND HOMEOSTATIC AGENTS

ESI学科分类：BIOLOGY & BIOCHEMISTRY

WOS号：WOS:001182536600001

JCR分区：Q4

影响因子：3.2

年份：2024

卷号：38

期号：3

开始页：2635

结束页：2649

文献类型：Article

关键词：HO-1; P62/RelA; osteosarcoma; apoptosis

摘要："Background: Heme oxygenase (HO)-1, a stress-induced heat shock protease, has been implicated in the occurrence and progression of various tumors, though its impact on osteosarcoma remains unclear. This study aimed to elucidate the role of the HO-1 gene in regulating apoptosis in osteosarcoma cells and its associated mechanism. Method: Initially, bioinformatics analysis was employed to analyze the expression of HO-1 in osteosarcoma. Subsequently, human osteosarcoma cells (MG63 and HOS cell lines) were cultured in vitro to establish osteosarcoma cells transfected with adenovirus overexpressing HO-1. Cell activity was assessed using cell counting kit-8 (CCK-8) and colony formation assays. Western blotting was utilized to analyze levels of HO-1, microtubule-associated proteins light chain 3 (LC3), recombinant nucleoporin 62 (P62), transcription factor P65 (P65), B-cell lymphoma-2 (Bcl-2), Bcl-2 assaciated X protein (Bax), cleaved caspase-3, beta-actin, and phosphorylation p65 (P65-RelA). Flow cytometry was conducted to examine distribution of cell cycle and cell apoptosis. Transmission electron microscopy (TEM) was employed to detect autophagy, while immunofluorescence was used to observe P65-RelA nuclear expression. Additionally, apoptosis of osteosarcoma cells was assessed via terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) staining. Knockdown of P62 and treatment with an nuclear factor-k-gene binding (NF-kappa B) inhibitor (PDTC) preceded Western blotting analysis, and apoptosis was evaluated using TUNEL staining. P65-RelA levels were assessed through immunofluorescence assay. Finally, a tumor-bearing nude mouse model was employed to validate the role of HO-1 in osteosarcoma in vivo. Results: The GSE11414 and GSE12865 chips were selected from the Gene Expression Omnibus (GEO) database and analyzed using the GEO 2R tool. The results indicated low expression of HO-1 in osteosarcoma. An in vitro osteosarcoma cell model with HO-1 overexpression was successfully established. Overexpression of HO-1 inhibited autophagic flow, promoted apoptosis in MG63 and HOS cells (p < 0.001), and reduced cell proliferation. Immunofluorescence analysis revealed an elevation in P65-RelA nuclear levels. Furthermore, transmission electron microscopy showed a significant reduction in the number of autophagic lysosomes. This phenomenon was reversed by pyrrolidine dithiocarbamate (PDTC) and siRNA-P62. Animal experiments confirmed the inhibitory effect of HO-1 on osteosarcoma. Conclusion: HO-1 is downregulated in osteosarcoma. HO-1 impairs autophagic flux in osteosarcoma cells, leading to the accumulation of P62, which activates apoptosis through the P62/RelA pathway and inhibits osteosarcoma tumorigenesis and progression."

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