Wang Sheng-Tao, Xu Shu-Juan, Gui Peng, Li Xin-Ning, Sui Yu-Han, Li Zhao-Xu
Department of Joint Surgery and Sports Medicine,Nanxishan Hospital of Guangxi Zhuang Autonomous Region, Guilin,Guangxi 541002,China.
Department of Hematology,Affiliated Hospital of Guilin Medical University,Guilin,Guangxi 541001,China.
Zhongguo Yi Xue Ke Xue Yuan Xue Bao. 2024 Feb;46(1):11-18. doi: 10.3881/j.issn.1000-503X.15746.
Objective To investigate the effect of staphylococcal nuclease and tudor domain containing 1(SND1) on the biological function of osteosarcoma cells and decipher the mechanism of SND1 in regulating ferroptosis in osteosarcoma cells SLC7A11. Methods Human osteoblasts hFOB1.19 and osteosarcoma cell lines Saos-2,U2OS,HOS,and 143B were cultured,in which the expression level of SND1 was determined.Small interfering RNA was employed to knock down the expression of SND1(si-SND1) in the osteosarcoma cell line HOS and 143B.The CCK8 assay kit,colony formation assay,and Transwell assay were employed to examine the effect of SND1 expression on the biological function of osteosarcoma cells.Furthermore,we altered the expression of SND1 and SLC7A11 in osteosarcoma cells to investigate the effect of SND1 on osteosarcoma ferroptosis SLC7A11. Results The mRNA and protein levels of SND1 in Saos-2,U2OS,HOS,and 143B cells were higher than those in hFOB1.19 cells(all <0.01).Compared with the control group,transfection with si-SND1 down-regulated the expression level of SND1 in HOS and 143B cells(all <0.01),decreased the viability of HOS and 143B cells,reduced the number of colony formation,and inhibited cell invasion and migration(all <0.001).The ferroptosis inducer Erastin promoted the apoptosis of HOS and 143B cells,while the ferroptosis inhibitor Ferrostatin-1 improved the viability of HOS and 143B cells(all <0.001).After SND-1 knockdown,Erastin reduced the viability of HOS and 143B cells,while Ferrostatin-1 restored the cell viability(all <0.001).After treatment with Erastin in the si-SND1 group,the levels of iron and malondialdehyde were elevated,and the level of glutathione was lowered(all <0.001).The results of experiments showed that SND1 knockdown inhibited the mass of the transplanted tumor in 143B tumor-bearing nude mice(<0.001).Knocking down the expression of SND1 resulted in down-regulated SLC7A11 expression(all <0.001) and increased ferroptosis in HOS and 143B cells(<0.001,=0.020). Conclusions SND1 presents up-regulated expression in osteosarcoma cells.It may inhibit ferroptosis by up-regulating the expression of SLC7A11,thereby improving the viability of osteosarcoma cells.
目的 探讨含葡萄球菌核酸酶和 Tudor 结构域 1（SND1）对骨肉瘤细胞生物学功能的影响，并阐明 SND1 在调控骨肉瘤细胞铁死亡中的机制及溶质载体家族 7 成员 11（SLC7A11）的作用。方法 培养人成骨细胞 hFOB1.19 及骨肉瘤细胞系 Saos-2、U2OS、HOS 和 143B，检测 SND1 的表达水平。采用小干扰 RNA 敲低骨肉瘤细胞系 HOS 和 143B 中 SND1（si-SND1）的表达。使用 CCK8 检测试剂盒、集落形成实验和 Transwell 实验检测 SND1 表达对骨肉瘤细胞生物学功能的影响。此外，改变骨肉瘤细胞中 SND1 和 SLC7A11 的表达，研究 SND1 对骨肉瘤细胞铁死亡及 SLC7A11 的影响。结果 Saos-2、U2OS、HOS 和 143B 细胞中 SND1 的 mRNA 和蛋白水平均高于 hFOB1.19 细胞（均 P<0.01）。与对照组相比，转染 si-SND1 可下调 HOS 和 143B 细胞中 SND1 的表达水平（均 P<0.01），降低 HOS 和 143B 细胞的活力，减少集落形成数量，并抑制细胞侵袭和迁移（均 P<0.001）。铁死亡诱导剂 Erastin 促进 HOS 和 143B 细胞凋亡，而铁死亡抑制剂 Ferrostatin-1 提高 HOS 和 143B 细胞的活力（均 P<0.001）。敲低 SND1 后，Erastin 降低 HOS 和 143B 细胞的活力，而 Ferrostatin-1 恢复细胞活力（均 P<0.001）。在 si-SND1 组中用 Erastin 处理后，铁和丙二醛水平升高，谷胱甘肽水平降低（均 P<0.001）。实验结果表明，敲低 SND1 可抑制 143B 荷瘤裸鼠移植瘤的质量（P<0.001）。敲低 SND1 的表达导致 SLC7A11 表达下调（均 P<0.001），并增加 HOS 和 143B 细胞的铁死亡（P<0.001，P = 0.020）。结论 SND1 在骨肉瘤细胞中呈上调表达。它可能通过上调 SLC7A11 的表达来抑制铁死亡，从而提高骨肉瘤细胞的活力。
