基于TAMs的极化探讨雷公藤内酯醇对耐药上皮性卵巢癌抑制机制的研究

The treatment of cisplatin ovarian cancer has become a thorny issue, and it is of great significance to explore the safe and effective anti-drug ovarian cancer. Our previous work found that tripterygium (TP) had a inhibitory effect on drug-resistant ovarian cancer, and studied its related inhibitory mechanism. Tumor associated macrophage (TAMs, tumor infiltrating leukocytes in the main component) can induce tumor growth, TAMs polarization in play an important role in the occurrence and development of malignant tumor, and PI3K/Akt signaling pathway in macrophage polarization plays an important role. Our previous work found that TP can block the PI3K/AKT signaling pathway and thus inhibit ovarian cancer. According to this assumption, TP can promote TAMs to m1-type polarization by PI3K/AKT signaling pathway, and inhibit its polarization to M2. Therefore, this research proposed from molecular level, cellular level, organization, and animal levels after multi-level observation of TP in tumor, polarization and TAMs PI3K/AKT signaling pathway related protein changes, so as to verify the TP through regulating tumor microenvironment change and adjustment of PI3K/AKT signal pathway and thus play a role of anti-tumor. This new viewpoint provides experimental basis and theoretical basis for the treatment of patients with chemotherapy drug ovarian cancer.

耐顺铂卵巢癌的治疗已成为棘手问题，挖掘安全有效的抗耐药卵巢癌中药意义重大。我们前期工作发现雷公藤内酯醇（TP)对耐药卵巢癌有抑制作用，并研究了其相关抑制机制。肿瘤相关巨噬细胞（TAMs，肿瘤中浸润白细胞的主要成分）可诱导肿瘤的生长，TAMs的极化在恶性肿瘤的发生、发展中起重要作用，而PI3K/Akt信号通路在巨噬细胞极化扮演重要的角色。我们前期工作发现，TP可阻断PI3K/AKT信号通路进而抑制卵巢癌。据此假设：TP可通过PI3K/AKT信号通路促进TAMs向M1型极化，抑制其向M2型极化。因此，本课题组拟从分子水平、细胞水平、组织以及动物水平等多层次观察TP作用于肿瘤后，TAMs极化情况及PI3K/AKT信号通路相关蛋白的变化，从而验证TP通过调控肿瘤微环境改变及对PI3K/AKT信号通路调节进而发挥抗肿瘤作用。这个新视点为TP为耐化疗药物卵巢癌患者的治疗提供实验依据和理论基础.

耐顺铂和晚期卵巢癌的治疗已成为目前临床医师治疗上棘手问题，寻找安全有效的抗耐顺铂化疗药物的卵巢癌中药意义重大。我们前期工作发现雷公藤内酯醇（TP)对耐顺铂卵巢癌有抑制作用，并研究了其相关抑制机制。肿瘤相关巨噬细胞（TAMs）可诱导恶性肿瘤的生长，TAMs的极化在恶性肿瘤的发生、发展中起重要作用，而PI3K/Akt/NF-κB信号通路在TAMs极化扮演着重要的角色。我们前期工作也发现，TP可阻断PI3K/AKT信号通路进而抑制耐顺铂人上皮性卵巢癌生长。本研究通过诱导巨噬细胞，采用共培养模型模拟肿瘤微环境，观察不同剂量TP对共培养体系的影响，以研究肿瘤细胞的生物学行为。建立体内、外实验明确 TP 干预对 M2 巨噬细胞肿瘤微环境浸润的影响及 MMP2 / MMP9 /VEGF 表达的影响。观察 PI3K/AKT 信号通路在 TP 干预后 M2 巨噬细胞极化的变化。实验结果提示：1、体外诱导TAMs可行。2、TP可抑制A2780/DDP细胞生长，促进细胞凋亡，呈时间-剂量依赖关系。3、TP可协同DDP诱导人耐顺铂卵巢癌A2780/DDP 细胞凋亡，增加A2780/DDP对顺铂的敏感性。TP诱导肿瘤细胞凋亡机制可能与抑制了PI3K/AKT/NF-κB 生存信号通路，下调磷酸化AKT和磷酸化NF-κB表达，抑制MMP-2、MMP-9及VEGF蛋白表达，从而抑制TAMs向M2型巨噬细胞极化，促进TAMs向M1型巨噬细胞极化，促进卵巢癌细胞凋亡。从而验证了TP通过对PI3K/AKT/NF-κB信号通路调节及调控肿瘤微环境进而发挥抗肿瘤作用。这个新视点为TP为耐顺铂化疗药物的上皮性卵巢癌和晚期卵巢癌患者的治疗提供实验依据和理论基础。