磷酸酶Wip1协同HPV E6癌蛋白在宫颈癌新辅助化疗铂类敏感性中的作用及机制研究

Cervical cancer is the most common malignant tumor of the female reproductive system. The patient's insensitivity to the current conventional chemotherapy regimen seriously affects the prognosis. The specific regulation mechanism of cervical cancer patients' insensitivity to platinum-based chemotherapy drugs is still unclear, which greatly restricts the progress of individualized treatment of cervical cancer.Our preliminary study found that HPV E6 oncoprotein and phosphatase Wip1 were highly expressed in cancer tissues with no response to neoadjuvant chemotherapy in cervical cancer, and they could significantly reduce the sensitivity of cervical cancer cells to cisplatin in vitro. Based on the in vivo and in vitro model, RNA-seq, multi-color immunohistochemistry, CRISPR-Cas9, 3D microfluidic culture technology and small molecule inhibitor technology, this project aims to clarify the synergistic effect of HPV E6-Wip1 in platinum-based chemotherapy sensitivity of cervical cance, and find the downstream signaling pathways and key molecules which could predict chemosensitivity. The purpose is to systematically reveal the regulation mechanism of Wip1 combined with HPV E6 in cervical cancer platinum-based chemotherapy, discover some sensitive indicators for predicting chemotherapy sensitivity, and provide theoretical basis for individualized treatment and improving prognosis.

宫颈癌是最常见的女性生殖系统恶性肿瘤。癌症患者对化疗药物是否与预后密切相关。目前患者对铂类化疗药物敏感性的具体调控机制仍不明确，极大制约了宫颈癌个体化治疗方案进展。前期研究首次发现HPV E6癌蛋白和磷酸酶Wip1在宫颈癌新辅助化疗无反应的癌组织中高表达，体外均能显著降低宫颈癌细胞对顺铂的敏感性。本项目拟在前期研究的基础上，利用RNA-seq、多色免疫组织化学、CRISPR-Cas9、3D微流体培养、蛋白质组学、小分子抑制剂、小鼠皮下瘤和PDX模型等技术，明确HPV E6-Wip1在宫颈癌铂类化疗敏感性中的协同作用模式，发现下游信号通路及预测化疗敏感性的关键分子。旨在系统揭示Wip1联合HPV E6调控宫颈癌铂类化疗敏感性中的机制，为预测化疗耐药提供敏感性指标，并为宫颈癌个体化治疗和改善预后提供理论依据。

宫颈癌是女性生殖系统最常见恶性肿瘤。近年来，尽管宫颈癌的预防、筛查、诊断和治疗取得关键进展，转移和复发患者的总体预后仍旧很差。化疗耐药和转移是影响中晚期或复发性宫颈癌预后的关键因素和治疗的主要挑战，亦是当前癌症研究的热点和难点。本研究首次报道HPV E6癌蛋白调节Wip1磷酸酶表达，进而降低宫颈癌患者对NACT中铂类化疗药物的敏感性。通过转录组测序，在细胞水平发现HPV E6可以调控Wip1表达。本研究通过临床标本入手， 高表达HPV E6癌蛋白和Wip1的患者，预后较差。通过体外的流式细胞学技术、Western Blot技术、小分子抑制剂、3D微流体类器官培养技术及AO/PI技术，我们证实了Wip1可以调控下游的ATM-Chk1/Chk2及p38MAPK-ERK通路；Wip1抑制剂及下游相关分子抑制剂均可增强不同宫颈癌细胞对顺铂的敏感性。在体内实验我们证实Wip1抑制剂可以增强顺铂的杀伤作用，小鼠生存时间延长。综上所述，本研究指出：Wip1在HPV E6介导的宫颈癌NACT过程中对铂类化疗药物的敏感性中关键作用及其潜在的机制，为宫颈癌患者预测化疗耐药提供敏感性指标，并为宫颈癌个体化治疗和改善预后提供理论依据。

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