Department of Orthopedics, The First Affiliated Hospital of Chongqing Medical University, No. 1 YouYi Road, Yuan Jia Gang, Yu Zhong District, Chongqing, 400016, People's Republic of China.
J Orthop Surg Res. 2023 Feb 27;18(1):145. doi: 10.1186/s13018-023-03572-4.
The treatment of osteosarcoma (OS) is still mainly surgery combined with systematic chemotherapy, and gene therapy is expected to improve the survival rate of patients. This study aimed to explore the effect of DEP domain 1 protein (DEPDC1) and kinesin super-family protein 4A (KIF4A) in OS and understand its mechanism. Th expression of DEPDC1 and KIF4A in OS cells was detected by RT-PCR and western blot. The viability, proliferation, invasion and migration of OS cells and tube formation of human umbilical vein endothelial cells (HUVECs) after indicated treatment were in turn detected by CCK-8 assay, EdU staining, wound healing assay, transwell assay and tube formation assay. The interaction between DEPDC1 and KIF4A was predicted by STRING and confirmed by co-immunoprecipitation. The expression of epithelial-mesenchymal transition (EMT)-related proteins, tube formation-related proteins and Hippo signaling pathway proteins was detected by western blot. As a result, the expression of DEPDC1 and KIF4A was all increased in U2OS cells. Down-regulation of DEPDC1 suppressed the viability, proliferation, invasion and migration of U2OS cells and tube formation of HUVECs, accompanied by the increased expression of E-cadherin and decreased expression of N-cadherin, Vimentin and VEGF. DEPDC1 was confirmed to be interacted with KIF4A. Upregulation of KIF4A partially reversed the effect of DEPDC1 interference on the above biological behaviors of U2OS cells. Down-regulation of DEPDC1 promoted the expression of p-LATS1 and p-YAP in Hippo signaling pathway, which was reversed by upregulation of KIF4A. In conclusion, down-regulation of DEPDC1 inhibited the malignant biological behavior of OS cells through the activation of Hippo signaling pathway, which could be reversed by upregulation of KIF4A.
骨肉瘤（OS）的治疗仍然主要是手术结合系统化疗，基因治疗有望提高患者的生存率。本研究旨在探讨 DEP 结构域 1 蛋白（DEPDC1）和驱动蛋白超家族蛋白 4A（KIF4A）在骨肉瘤中的作用，并了解其机制。通过 RT-PCR 和 Western blot 检测 OS 细胞中 DEPDC1 和 KIF4A 的表达。依次通过 CCK-8 测定、EdU 染色、划痕愈合测定、Transwell 测定和管形成测定检测指示处理后 OS 细胞的活力、增殖、侵袭和迁移以及人脐静脉内皮细胞（HUVECs）的管形成。通过 STRING 预测 DEPDC1 和 KIF4A 之间的相互作用，并通过共免疫沉淀进行验证。通过 Western blot 检测上皮-间充质转化（EMT）相关蛋白、管形成相关蛋白和 Hippo 信号通路蛋白的表达。结果，U2OS 细胞中 DEPDC1 和 KIF4A 的表达均增加。下调 DEPDC1 抑制 U2OS 细胞的活力、增殖、侵袭和迁移以及 HUVECs 的管形成，同时 E-钙粘蛋白表达增加，N-钙粘蛋白、波形蛋白和 VEGF 表达减少。证实 DEPDC1 与 KIF4A 相互作用。上调 KIF4A 部分逆转了 DEPDC1 干扰对 U2OS 细胞上述生物学行为的影响。下调 DEPDC1 促进 Hippo 信号通路中 p-LATS1 和 p-YAP 的表达，上调 KIF4A 可逆转这一作用。总之，下调 DEPDC1 通过激活 Hippo 信号通路抑制骨肉瘤细胞的恶性生物学行为，上调 KIF4A 可逆转这一作用。
