School of Clinical Medicine, Guizhou Medical University, Guiyang 550000, Guizhou, China.
Department of Orthopedics, The Affiliated Hospital of Guizhou Medical University, Guiyang 550000, Guizhou, China.
Aging (Albany NY). 2024 Jun 17;16(12):10462-10476. doi: 10.18632/aging.205941.
Osteosarcoma is a highly malignant bone tumor that exhibits rapid growth and early metastasis. Hypoxia plays a pivotal role in promoting the proliferation and metastasis of osteosarcoma through a series of molecular events, which are partially mediated and regulated by HIF-1α. However, the regulatory network associated with HIF-1α in osteosarcoma remains limited. Therefore, the objective of this study was to identify critical hypoxia-associated genes and investigate their effects and molecular mechanisms in osteosarcoma cells.
Through bioinformatics analysis, matrilin-4 (MATN4) was identified as a crucial gene associated with hypoxia. The expression of MATN4 and HIF-1α was assessed using immunohistochemistry, RT-qPCR, and western blotting. The proliferative capacity of osteosarcoma cells was assessed through the utilization of CCK-8, EDU staining, and colony formation assays. The effects of MATN4 on the mobility of OS cells were evaluated using wound-healing assays and transwell assays. The interaction between MATN4 and HIF-1α was detected through chromatin immunoprecipitation.
MATN4 is overexpressed in osteosarcoma tissue and cells, particularly in osteosarcoma cells with high metastatic potential. Knockdown of MATN4 inhibits the proliferation, migration, and invasion abilities of osteosarcoma cells and reverses the promoting effects of hypoxia on these functions. Additionally, HIF-1α binds to MATN4 and upregulates its expression. Interestingly, knockdown of HIF-1α reduces the stimulatory effects of MATN4 overexpression on the proliferation, migration, and invasion of osteosarcoma cells under hypoxic conditions.
Taken together, our results suggest that MATN4 is regulated by HIF-1α and confers a more aggressive phenotype on OS cells. This evidence suggests that MATN4 may act as a potential target for OS diagnosis and treatment.
骨肉瘤是一种高度恶性的骨肿瘤，具有快速生长和早期转移的特点。缺氧通过一系列分子事件在促进骨肉瘤的增殖和转移中起着关键作用，这些事件部分由 HIF-1α 介导和调节。然而，骨肉瘤中与 HIF-1α 相关的调控网络仍然有限。因此，本研究的目的是鉴定关键的缺氧相关基因，并研究它们在骨肉瘤细胞中的作用和分子机制。
通过生物信息学分析，鉴定出连接蛋白 4（MATN4）是与缺氧相关的关键基因。采用免疫组织化学、RT-qPCR 和 Western blot 检测 MATN4 和 HIF-1α 的表达。通过 CCK-8、EDU 染色和集落形成实验评估骨肉瘤细胞的增殖能力。通过划痕愈合实验和 Transwell 实验评估 MATN4 对 OS 细胞迁移能力的影响。通过染色质免疫沉淀检测 MATN4 与 HIF-1α 之间的相互作用。
MATN4 在骨肉瘤组织和细胞中表达上调，特别是在具有高转移潜能的骨肉瘤细胞中。MATN4 敲低抑制了骨肉瘤细胞的增殖、迁移和侵袭能力，并逆转了缺氧对这些功能的促进作用。此外，HIF-1α 与 MATN4 结合并上调其表达。有趣的是，HIF-1α 敲低降低了 MATN4 过表达在缺氧条件下对骨肉瘤细胞增殖、迁移和侵袭的刺激作用。
综上所述，我们的研究结果表明，MATN4 受 HIF-1α 调节，并赋予 OS 细胞更具侵袭性的表型。这一证据表明，MATN4 可能成为骨肉瘤诊断和治疗的潜在靶点。
