Ovarian cancer is the most malignant gynecologic cancer. Previous studies found that lamin-A was associated with DNA damage repair proteins but the underlying mechanism remains unclear. We speculate that this may be related to its interacting proteins, such as Hsp90. The aim of this study is to investigate the effects of Hsp90 on DNA damage repair and chemoresistance of ovarian cancer cells. In our research, co-immunoprecipitation (co-IP) and mass spectrometry (MS) were used to identify proteins interacting with lamin-A and the interaction domain. Next, the relationship between lamin-A and Hsp90 was explored by Western blotting (WB) and immunofluorescence staining. Then, effect of Hsp90 inhibition on DNA damage repair was assessed through detecting Rad50 and Ku80 by WB. Furthermore, to test the roles of 17-AAG on cell chemosensitivity, CCK-8 and colony formation assay were carried out. Meanwhile, IC50 of cells were calculated, followed by immunofluorescence to detect DNA damage. At last, the mouse xenograft model was used in determining the capacity of 17-AAG and DDP to suppress tumor growth and metastatic potential. The results showed that lamin-A could interact with Hsp90 via the domain of lamin-A^1-430. Besides, the distribution of Hsp90 could be affected by lamin-A. After lamin-A knockdown, Hsp90 decreased in the cytoplasm and increased in the nucleus, suggesting that the interaction between lamin-A and Hsp90 may be related to the nucleocytoplasmic transport of Hsp90. Moreover, inhibition of Hsp90 led to an obvious decrease in the expression of DSBs (DNA double-strand break) repair proteins, as well as cell proliferation ability upon DDP treatment and IC50 of DDP, causing more serious DNA damage. In addition, the combination of 17-AAG and DDP restrained the growth of ovarian cancer efficiently in vivo and prolonged the survival time of tumor-bearing mice.

卵巢癌是恶性程度最高的妇科癌症。先前的研究发现，lamin-A 与 DNA 损伤修复蛋白有关，但其潜在机制仍不清楚。我们推测这可能与其相互作用的蛋白质有关，例如 Hsp90。本研究的目的是研究 Hsp90 对卵巢癌细胞 DNA 损伤修复和化学抗性的影响。在我们的研究中，共免疫沉淀 (co-IP) 和质谱 (MS) 用于识别与 lamin-A 和相互作用域相互作用的蛋白质。接下来，通过蛋白质印迹（WB）和免疫荧光染色探索了lamin-A和Hsp90之间的关系。然后，通过WB检测Rad50和Ku80来评估Hsp90抑制对DNA损伤修复的影响。此外，为了测试 17-AAG 对细胞化学敏感性的作用，进行CCK-8和菌落形成测定。同时计算细胞的IC50，免疫荧光检测DNA损伤。最后，小鼠异种移植模型被用于确定17-AAG和DDP抑制肿瘤生长和转移潜能的能力。结果表明，lamin-A 可以通过 lamin-A 的结构域与 Hsp90 相互作用。^1-430。此外，Hsp90 的分布可能受 lamin-A 的影响。在lamin-A敲低后，Hsp90在细胞质中减少而在细胞核中增加，表明lamin-A和Hsp90之间的相互作用可能与Hsp90的核质转运有关。此外，抑制Hsp90导致DSBs（DNA双链断裂）修复蛋白的表达明显下降，DDP处理后的细胞增殖能力和DDP的IC50下降，导致更严重的DNA损伤。此外，17-AAG 和 DDP 的组合在体内有效地抑制了卵巢癌的生长，并延长了荷瘤小鼠的存活时间。