卵巢癌的死亡率很高，主要是因为它对目前使用的疗法有抵抗力。这种耐药性与癌症干细胞 (CSC) 的存在、与微环境的相互作用以及瘤内异质性有关。因此，寻找新的治疗靶点，特别是针对CSC的治疗靶点，对于改善患者预后非常重要。已发现HOOK1在相当大比例的卵巢肿瘤中发生转录改变，但其在肿瘤发生和发展中的作用仍不完全清楚。使用CRISPR/Cas9技术在卵巢癌细胞系中下调HOOK1，然后生长体外和体内测定。随后，对下调细胞系的迁移（博伊登室）、细胞死亡（蛋白质印迹和流式细胞术）和干性特性（克隆异质性分析、肿瘤球测定和流式细胞术）进行了分析。为了深入了解 HOOK1 在卵巢癌中的具体作用机制，进行了蛋白质组分析，然后进行了蛋白质印迹和细胞毒性测定，以确认质谱中发现的结果。免疫荧光染色、蛋白质印迹和流式细胞术也被用来完成揭示HOOK1在卵巢癌中的作用。在这项研究中，我们观察到降低卵巢癌细胞中HOOK1的水平可减少体外生长和迁移，并阻止卵巢癌细胞中肿瘤的形成。体内。此外，HOOK1 的减少导致这些细胞中干细胞样能力的下降，但这似乎与传统上与这种表型相关的基因的表达无关。蛋白质组研究以及其他分析表明，HOOK1 的下调还诱导这些细胞内质网应激水平的增加。最后，在 HOOK1 下调的细胞中观察到的干细胞样特性的下降可以解释为，由于未折叠蛋白反应引起的内质网应激，培养物中 CSC 群体的细胞死亡增加。HOOK1 有助于维持致瘤性和干性。通过保持蛋白质稳态来改变卵巢癌细胞的特性，可被视为替代治疗靶点，特别是与内质网诱导剂或蛋白酶体抑制剂等蛋白毒性应激剂联合使用。© 2024。作者。

Ovarian cancer has a high mortality rate mainly due to its resistance to currently used therapies. This resistance has been associated with the presence of cancer stem cells (CSCs), interactions with the microenvironment, and intratumoral heterogeneity. Therefore, the search for new therapeutic targets, particularly those targeting CSCs, is important for improving patient prognosis. HOOK1 has been found to be transcriptionally altered in a substantial percentage of ovarian tumors, but its role in tumor initiation and development is still not fully understood.The downregulation of HOOK1 was performed in ovarian cancer cell lines using CRISPR/Cas9 technology, followed by growth in vitro and in vivo assays. Subsequently, migration (Boyden chamber), cell death (Western-Blot and flow cytometry) and stemness properties (clonal heterogeneity analysis, tumorspheres assay and flow cytometry) of the downregulated cell lines were analysed. To gain insights into the specific mechanisms of action of HOOK1 in ovarian cancer, a proteomic analysis was performed, followed by Western-blot and cytotoxicity assays to confirm the results found within the mass spectrometry. Immunofluorescence staining, Western-blotting and flow cytometry were also employed to finish uncovering the role of HOOK1 in ovarian cancer.In this study, we observed that reducing the levels of HOOK1 in ovarian cancer cells reduced in vitro growth and migration and prevented tumor formation in vivo. Furthermore, HOOK1 reduction led to a decrease in stem-like capabilities in these cells, which, however, did not seem related to the expression of genes traditionally associated with this phenotype. A proteome study, along with other analysis, showed that the downregulation of HOOK1 also induced an increase in endoplasmic reticulum stress levels in these cells. Finally, the decrease in stem-like properties observed in cells with downregulated HOOK1 could be explained by an increase in cell death in the CSC population within the culture due to endoplasmic reticulum stress by the unfolded protein response.HOOK1 contributes to maintaining the tumorigenic and stemness properties of ovarian cancer cells by preserving protein homeostasis and could be considered an alternative therapeutic target, especially in combination with inducers of endoplasmic reticulum or proteotoxic stress such as proteasome inhibitors.© 2024. The Author(s).