Derpartment of Orthopedics, Shaoguan First People's Hospital Affiliated to Southern Medical University, Guangdong, China.
Orthopedics Center, Zhujiang Hospital of Southern Medical University, Guangzhou, China.
Cancer Res Treat. 2022 Jan;54(1):277-293. doi: 10.4143/crt.2021.320. Epub 2021 May 11.
Osteosarcoma (OS) universally exhibits heterogeneity and cisplatin (CDDP) resistance. Although the Wee1/CDC2 and nuclear factor кB (NF-κB) pathways were reported to show abnormal activation in some tumor cells with CDDP resistance, whether there is any concrete connection is currently unclear. We explored it in human OS cells.
Multiple OS cell lines were exposed to a Wee1 inhibitor (AZD1775) and CDDP to assess the half-maximal inhibitory concentration values. Western blot, coimmunoprecipitation, confocal immunofluorescence, cell cycle, and Cell Counting Kit-8assays were performed to explore the connection between the Wee1/CDC2 and NF-κB pathways and their subsequent physiological contribution to CDDP resistance. Finally, CDDP-resistant PDX-OS xenograft models were established to confirm that AZD1775 restores the antitumor effects of CDDP.
A sensitivity hierarchy of OS cells to CDDP and AZD1775 exists. In the highly CDDP-tolerant cell lines, Wee1 and RelA were physically crosslinked, which resulted in increased abundance of phosphorylated CDC2 (Y15) and RelA (S536) and consequent modulation of cell cycle progression, survival, and proliferation. Wee1 inhibition restored the effects of CDDP on these processes in CDDP-resistant OS cells. In addition, animal experiments with CDDP-resistant PDX-OS cells showed that AZD1775 combined with CDDP not only restored CDDP efficacy but also amplified AZD1775 in inhibiting tumor growth and prolonged the median survival of the mice.
Simultaneous enrichment of molecules in the Wee1/CDC2 and NF-κB pathways and their consequent coactivation is a new molecular mechanism of CDDP resistance in OS cells. OS with this molecular signature may respond well to Wee1 inhibition as an alternative treatment strategy.
骨肉瘤（OS）普遍表现出异质性和顺铂（CDDP）耐药性。虽然已有报道称，在一些具有 CDDP 耐药性的肿瘤细胞中，Wee1/CDC2 和核因子 кB（NF-κB）途径表现出异常激活，但目前尚不清楚它们之间是否存在具体联系。本研究在人骨肉瘤细胞中对此进行了探讨。
将多种骨肉瘤细胞系暴露于 Wee1 抑制剂（AZD1775）和 CDDP 中，以评估半抑制浓度值。通过 Western blot、共免疫沉淀、共聚焦免疫荧光、细胞周期和细胞计数试剂盒-8 检测，探讨了 Wee1/CDC2 和 NF-κB 途径之间的联系及其对 CDDP 耐药性的后续生理贡献。最后，建立了 CDDP 耐药性 PDX-OS 异种移植模型，以证实 AZD1775 恢复了 CDDP 的抗肿瘤作用。
OS 细胞对 CDDP 和 AZD1775 的敏感性存在一个层次结构。在高度耐受 CDDP 的细胞系中，Wee1 和 RelA 发生物理交联，导致磷酸化 CDC2（Y15）和 RelA（S536）的含量增加，从而改变细胞周期进程、存活和增殖。Wee1 抑制恢复了 CDDP 对 CDDP 耐药性 OS 细胞中这些过程的作用。此外，用 CDDP 耐药性 PDX-OS 细胞进行的动物实验表明，AZD1775 联合 CDDP 不仅恢复了 CDDP 的疗效，而且增强了 AZD1775 抑制肿瘤生长的作用，并延长了小鼠的中位生存期。
Wee1/CDC2 和 NF-κB 途径中分子的同时富集及其随后的共激活是 OS 细胞中 CDDP 耐药性的一个新分子机制。具有这种分子特征的 OS 可能对作为替代治疗策略的 Wee1 抑制反应良好。
