SAR1A regulates the RhoA/YAP and autophagy signaling pathways to influence osteosarcoma invasion and metastasis

• Published in: Cancer science 2022-12, Vol.113 (12), p.4104-4119
• Main Authors: Zhan, Fangbiao, Deng, Qianrong, Chen, Zhiyu, Xie, Chaozheng, Xiang, Shuang, Qiu, Sheng, Tian, Lin, Wu, Chunrong, Ou, Yunsheng, Chen, Jian, Xu, Lixin
• Format: Article
• Language: English
• Subjects: Adolescent; Autophagy; Autophagy - genetics; Bioinformatics; Bone cancer; Bone Neoplasms - pathology; Cell Line, Tumor; Cell migration; Cell Movement - genetics; Cell Proliferation; Endoplasmic reticulum; ER stress; Gene expression; Homeostasis; Humans; Kinases; Lamellipodia; Lung Neoplasms - genetics; Lung Neoplasms - secondary; Malignancy; Mesenchyme; Metastases; Metastasis; Microscopy; Monomeric GTP-Binding Proteins - metabolism; Original; Osteosarcoma; Osteosarcoma - pathology; Osteosarcoma cells; Patients; Phalloidin; Proteins; Pseudopodia; ras Proteins - metabolism; Reactive oxygen species; rhoA GTP-Binding Protein - genetics; rhoA GTP-Binding Protein - metabolism; RhoA protein; RhoA/YAP; SAR1A; Signal Transduction; Tumors; Western blotting; Wound healing; Yes-associated protein
• ISSN: 1347-9032; 1349-7006
• DOI: 10.1111/cas.15551

Osteosarcoma is the most prevalent form of primary bone malignancy affecting adolescents. Secretion‐associated Ras‐related GTPase 1A (SAR1A) is a key regulator of endoplasmic reticulum (ER) homeostasis, but its role as a regulator of osteosarcoma metastasis has yet to be clarified. Bioinformatics analyses revealed SAR1A and RHOA to be upregulated in osteosarcoma patients, with the upregulation of these genes being associated with poor 5‐year metastasis‐free survival rates. In addition, the upregulation of SAR1A and RHOA in osteosarcoma was highly positively correlated. Immunohistochemical analyses additionally revealed that SAR1A levels were increased in osteosarcoma pulmonary metastases. In vitro wound healing and Transwell assays indicated that knocking down SAR1A or RHOA impaired the invasive and migratory activity of osteosarcoma cells, whereas RHOA overexpression had the opposite effect. Western blotting and immunofluorescent staining revealed the inhibition of osteosarcoma cell epithelial–mesenchymal transition following SAR1A or RHOA knockdown; RHOA overexpression had the opposite effect. Following SAR1A knockdown, phalloidin staining indicated that osteosarcoma cells showed reduced lamellipodia formation. Endoplasmic reticulum stress levels and reactive oxygen species production were enhanced following the knockdown of SAR1A, as was autophagic activity, with lung metastases being reduced in vivo after such knockdown. Knocking down SAR1A suppresses osteosarcoma cell metastasis through the RhoA/YAP, ER stress, and autophagic pathways, offering new insights into the regulation of autophagic activity in the context of osteosarcoma cell metastasis and suggesting that these pathways could be amenable to therapeutic intervention. These results offer novel evidence that SAR1A can function in manner to promote metastasis and to regulate autophagic activity. Knocking down SAR1A can prevent metastasis by modulating the RhoA/YAP, ER stress, and autophagic activities, offering new insight into the mechanisms whereby intratumoral autophagy is regulated and highlighting these pathways as promising targets for therapeutic intervention in osteosarcoma.