Department of Orthopaedic, Shanghai Tenth People's Hospital, School of Medicine, Tongji University, Shanghai, China.
Drug Dev Res. 2023 Dec;84(8):1751-1763. doi: 10.1002/ddr.22120. Epub 2023 Oct 2.
Osteosarcoma is a prevalent malignant bone tumor with a poor prognosis. Mothers against decapentaplegic homolog 3 (Smad3) present as a therapeutic target in antitumor treatment, whereas its functions in the osteosarcoma have not been well explored. In the current study, we aimed to investigate the effects of Smad3 in the progression of osteosarcoma. The tumor immune single-cell hub 2 website was used for graph-based visualization of Smad3 status in osteosarcoma single-cell database. Western Blot was applied to detect the expression of Smad3 protein in cell lines. Colony formation and cell counting kit-8 assays were used to evaluate cell proliferation. Transwell and wound healing assays were used to detect the migration and invasion abilities of cells. Cell apoptosis rates and cell cycle changes were explored by using flow cytometry analysis. The xenograft tumor growth model was applied to explore the effect in tumor growth after Smad3 blockage in vivo. Moreover, to confirm the potential mechanism of Smad3's effects on osteosarcoma, bioinformatics analysis was performed in TARGET-Osteosarcoma and GSE19276 databases. Our study found that the Smad3 protein is overexpressed in 143B and U2OS cells, suppressing the expression of Smad3 protein in osteosarcoma cells by Smad3 target inhibitor (E)-SIS3 or lentivirus can inhibit the proliferation, migration, invasion, promote cell apoptosis, arrest cell G1 cycle in osteosarcoma cells in vitro, and suppress tumor growth in vivo. Furthermore, the bioinformatics analysis demonstrated that high expression of Smad3 is closely associated with low immune status in TARGET-Osteosarcoma and GSE19276 databases. Our study suggested that Smad3 could contribute positively to osteosarcoma progression via the regulation of tumor immune microenvironment, and Smad3 may represent as an valuable potential therapeutic target in osteosarcoma therapy.
骨肉瘤是一种预后不良的常见恶性骨肿瘤。母亲对抗 Decapentaplegic 同源物 3（Smad3）作为抗肿瘤治疗的治疗靶点，但其在骨肉瘤中的作用尚未得到充分探索。在本研究中，我们旨在研究 Smad3 在骨肉瘤进展中的作用。肿瘤免疫单细胞集线器 2 网站用于基于图的可视化骨肉瘤单细胞数据库中的 Smad3 状态。Western blot 用于检测细胞系中 Smad3 蛋白的表达。集落形成和细胞计数试剂盒-8 测定用于评估细胞增殖。Transwell 和划痕愈合测定用于检测细胞的迁移和侵袭能力。通过流式细胞术分析探索细胞凋亡率和细胞周期变化。异种移植肿瘤生长模型用于研究体内阻断 Smad3 后对肿瘤生长的影响。此外，为了证实 Smad3 对骨肉瘤影响的潜在机制，在 TARGET-Osteosarcoma 和 GSE19276 数据库中进行了生物信息学分析。我们的研究发现，Smad3 蛋白在 143B 和 U2OS 细胞中过度表达，通过 Smad3 靶标抑制剂（E）-SIS3 或慢病毒抑制骨肉瘤细胞中 Smad3 蛋白的表达，可抑制骨肉瘤细胞的增殖、迁移、侵袭，促进细胞凋亡，体外阻滞细胞 G1 周期，并抑制体内肿瘤生长。此外，生物信息学分析表明，TARGET-Osteosarcoma 和 GSE19276 数据库中 Smad3 表达水平高与免疫状态低密切相关。我们的研究表明，Smad3 可通过调节肿瘤免疫微环境正向促进骨肉瘤进展，Smad3 可能成为骨肉瘤治疗的有价值的潜在治疗靶点。
