从单核巨噬细胞近分泌通路研究古法阳和汤治疗乳腺癌的生物学机制

According to ancient writings, Clinical curative effect of Yanghe Decotion for the treatment of breast cancer is better. For the past few years, clinical application of Yanghe Decotion in cancer treatment has been reported. However, the mechanism is poorly understood. Juxtacrine signalling from monocytes and macrophages plays an important role in the development and progression of tumor. Previous studies have found that Yanghe Decotion has obvious inhibitory effect on the pathway of anchor protein CD90. Accordingly, we put forward the hypothesis that Yanghe Decotion can intervene juxtacrine signalling from monocytes and macrophages to exert the anti-tumor efficiency by regulating the expression of CD90 and EphA4 in cancer stem cells. Based on the ancient applications of Yanghe Decotion curing breast cancer, we establish various cell models and monocytes and macrophage - breast cancer stem cell co-culture system, and explore and validate the targets and mechanism of Yanghe Decotion by using techniques such as qRT-PCR, shRNA and co-immunoprecipitation. According to the characteristics of the tumor development at different stages, corresponding animal models are established to verify that Yanghe Decotion inhibits tumor initiation, growth and metastasis by intervening juxtacrine signalling from monocytes and macrophages. Clarifying the modern biological mechanisms of Yanghe Decotion curing breast cancer and revealing principles and characteristics, this study is of great significance, moreover, it provides a new thinking for the antitumor mechanism of Yanghe Decotion.

古籍记载，阳和汤治疗乳腺癌临床疗效较好，近年来，阳和汤治疗肿瘤的临床应用也屡见报道，然其作用机制却不甚明了。单核巨噬细胞近分泌通路在肿瘤的发生发展过程中扮演了极其重要的角色，前期研究发现阳和汤对该通路锚定蛋白CD90的表达具有明显下调作用，据此提出假说认为阳和汤可通过调控肿瘤干细胞CD90和EphA4的表达，干预单核巨噬细胞近分泌信号转导通路，从而发挥抗肿瘤作用。本课题拟遵循古法阳和汤治疗乳腺癌的应用，建立多种细胞模型和单核巨噬细胞-乳腺癌干细胞共培养体系，采用qRT-PCR、shRNA、染色体免疫共沉淀等技术，探索并验证阳和汤的作用靶点及作用规律；并根据肿瘤发生发展不同阶段特点，建立相应动物模型，验证阳和汤通过干预单核巨噬细胞近分泌通路抑制肿瘤启动、生长及转移的作用。本研究对阐明古法阳和汤治疗乳腺癌的现代生物学机制，揭示其作用规律和特点具有重要意义，为阳和汤的抗肿瘤机制研究提供了新思路。

乳腺癌是女性最常见的恶性肿瘤之一。目前，对于该病的诊断和治疗已成为医学界的热点，随着现代科学技术的发展，对于乳腺癌的诊断已取得了长足进步，然而对于该病的治疗仍存在许多亟待解决的科学问题。本研究通过体外实验研究，探寻古法阳和汤干预单核巨噬细胞近分泌通路的作用靶点和作用规律，探索古法阳和汤治疗乳腺癌的作用机制；通过动物模型研究，验证古法阳和汤通过干预单核巨噬细胞近分泌通路抑制肿瘤启动、生长及转移的作用。相关实验研究结果表明阳和汤含药血清具有抑制MDA-MB-231细胞的增殖，且对细胞增殖的抑制作用呈时间与剂量依赖；阳和汤可诱导MDA-MB-231细胞凋亡，其作用机制可能与其抑制PI3K/Akt信号通路的活化有关；阳和汤能够降低乳腺癌细胞CD90的表达，从而阻碍单核巨噬细胞与乳腺癌干细胞的结合，阻断下游信号的传导；阳和汤可下调乳腺癌细胞MDA-MB-231中Smad3、NF-κB、IL-8、Bcl-2的表达，可能是阳和汤抑制乳腺癌进展的潜在机制；阳和汤具有显著抑制三阴性乳腺癌裸鼠移植瘤的生长和肿瘤细胞的增殖、转移的作用。实验结果还提示阳和汤可抑制乳腺癌干细胞HMLER90hi增殖，其作用机制可能与抑制单核巨噬细胞近分泌通路的传导有关。 本研究对阐明古法阳和汤治疗乳腺癌的现代生物学机制，揭示其作用规律和特点具有重要意义，同时也为阳和汤的抗肿瘤机制研究提供了新的思路。