PO.ET03.04 · 实验与分子治疗
靶向卵巢癌的化疗耐药
Targeting chemoresistance in ovarian cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
引言：卵巢癌（OvCa）是一种高度侵袭性的恶性肿瘤，以频繁复发和转移为特征，两者均导致其高死亡率。尽管大多数患者最初对铂类化疗有反应，但多达 80% 的患者最终产生耐药，使 OvCa 在很大程度上无法治愈。因此，迫切需要确定新的治疗靶点并制定克服化疗耐药的策略。本研究确定整合素 β4（ITGB4）为与顺铂耐药相关的关键分子驱动因子，并探讨其作为卵巢癌治疗靶点的潜力。
方法：使用 RNA 测序对顺铂敏感（WT）和顺铂耐药（CPR）的 OVCAR8 OvCa 球体进行转录组分析，以确定与化疗耐药相关的差异表达基因。通过 qRT-PCR 和 Western blot 验证选定的候选基因。使用公开数据集（GSE133859、TCGA-OV）以及对原发性和复发性 OvCa 患者样本组织微阵列的免疫荧光分析评估 ITGB4 的临床意义。使用基因操作（过表达和 CRISPR 介导的敲除）结合迁移、侵袭和顺铂敏感性实验检测 ITGB4 的功能作用。进行共免疫沉淀（Co-IP）以检测 ITGB4 与 TGFβ 受体 2（TGFβR2）之间的相互作用。
结果：转录组分析揭示 CPR 球体中 ITGB4 过表达和 SERPINB2 下调，这些发现与 GEO、TCGA-OV 数据集以及患者样本的 TMA 分析一致。铂耐药 OvCa 患者中高 ITGB4 表达与更差的总生存和无进展生存显著相关。在功能上，ITGB4 过表达增强了细胞侵袭、迁移和顺铂耐药，而 ITGB4 敲除使耐药细胞重新对顺铂敏感并抑制其侵袭表型。机制上，ITGB4 激活了 FAK 和 TGFβ/SMAD 信号通路，二者均为已知的 EMT 和化疗耐药介导因子。Co-IP 实验证明 ITGB4 与 TGFβR 之间存在直接相互作用，支持其在促进 SMAD3 激活中的协同作用。
结论：本研究确定 ITGB4 为卵巢癌顺铂耐药和侵袭性的核心调节因子，并强调 ITGB4 作为克服化疗耐药、改善 OvCa 患者治疗结局的有前景治疗靶点。
资助致谢：本工作由 COBRE P20GM135009、SCC CT Pilot Grant、PHF-Seed 和 PHF CTGA Grant Program 资助。
查看英文原文 English abstract
Introduction: Ovarian cancer (OvCa) is a highly aggressive malignancy characterized by frequent relapse and metastasis, both of which contribute to its high mortality rate. Although most patients initially respond to platinum-based chemotherapy, up to 80% eventually develop resistance, rendering OvCa largely incurable. Therefore, there is an urgent need to identify new therapeutic targets and develop strategies to overcome chemoresistance. This study identifies integrin beta4 (ITGB4) as a key molecular driver associated with cisplatin resistance and explores its potential as a therapeutic target in ovarian cancer.
Methods: Transcriptomic profiling of cisplatin-sensitive (WT) and cisplatin-resistant (CPR) OVCAR8 OvCa spheroids was performed using RNA sequencing to identify differentially expressed genes associated with chemoresistance. Selected candidates were validated by qRT-PCR, and western blotting. The clinical significance of ITGB4 was assessed using publicly available datasets (GSE133859, TCGA-OV) and immunofluorescence analysis of tissue microarray from primary and recurrent OvCa patient samples. Functional roles of ITGB4 were examined using genetic manipulations (overexpression and CRISPR-mediated knockout) followed by migration, invasion, and cisplatin sensitivity assays. Co-immunoprecipitation (Co-IP) was performed to examine the interaction between ITGB4 and TGFbeta receptor 2 (TGFbetaR2).
Results: Transcriptomic profiling revealed ITGB4 overexpression and SERPINB2 downregulation in CPR spheroids, findings that were consistent with GEO, TCGA-OV datasets as well as TMA analysis of patient samples. High ITGB4 expression in platinum-resistant OvCa patients was significantly associated with worse overall and progression-free survival. Functionally, ITGB4 overexpression enhanced cell invasion, migration, and cisplatin resistance, while ITGB4 knockout resensitized resistant cells to cisplatin and suppressed their invasive phenotype. Mechanistically, ITGB4 activated FAK and TGFbeta/SMAD signaling pathways, both of which are known mediators of EMT and chemoresistance. CO-IP assays demonstrated a direct interaction between ITGB4 and TGFbetaR, supporting a cooperative role in promoting SMAD3 activation.
Conclusion: This study identifies ITGB4 as a central regulator of cisplatin resistance and aggressiveness in ovarian cancer and highlights ITGB4 as a promising therapeutic target to overcome chemoresistance and improve treatment outcomes in OvCa patients.
Acknowledgment of Funding: The work is supported by COBRE P20GM135009, SCC CT Pilot Grant, PHF-Seed and PHF CTGA Grant Program.
利益披露 Disclosure
D. Penta, None..
V. Chandra, None..
L. Dockery, None..
R. Rai, None.