基于CSF-1/CSF-1R通路调节肝癌微环境中M2型巨噬细胞的迁移探究解毒颗粒预防肝癌术后复发转移机制

The postoperative recurrence and metastasis of liver cancer seriously affect survival.High level clinical studies have confirmed Jiedu granule markedly reduces the postoperative recurrence and prolongs survival of liver cancer. However, the underlying mechanism is unknown. Because primary liver cancer is an inflammation-associated tumor, the infiltration of M2 macrophage in the liver cancer microenvironment is closely related to the postoperative recurrence and metastasis. I previously found that Jiedu granule can obviously reduce the postoperative metastasis and M2 macrophage in adjacent tissue of rat with liver cancer. Jiedu granule maybe reduce M2 macrophage in adjacent tissue by inhibiting macrophage polarization to M2 or reducing M2 macrophage migration to adjacent tissue.CSF-1/CSF-1R signaling pathway is most likely just the important pathway to adjust M2 macrophages infiltration.We hypothesized that Jiedu granule prevents recurrence or metastasis of liver cancer by inhibiting M2 macrophage migration to adjacent tissue..Therefore, in this present research, we will prove whether the Jiedu granule inhibits M2 macrophage migrate to adjacent tissue, by establishing rat model after resection of liver cancer and co-culture system of liver cancer cells and M2 macrophages, through the technology of immunohistochemistry, PCR, Transwell, scratch test. We will explore whethere Jiedu granule inhibits M2 macrophage migrate to adjacent tissue by adjusting CSF-1/CSF-1R pathway.

肝癌术后复发转移严重影响患者生存。高级别临床研究证实解毒颗粒显著降低肝癌术后复发、延长生存期，但作用机制尚不明确。肝癌为炎症相关肿瘤，其微环境中M2巨噬细胞的浸润与肝癌复发转移密切相关。申请人预实验发现，解毒颗粒能明显减少大鼠肝癌旁组织中M2巨噬细胞浸润、降低术后转移。解毒颗粒可能通过抑制巨噬细胞向M2型极化及减少M2巨噬细胞向癌旁迁移，从而减少癌旁组织中M2巨噬细胞浸润。CSF-1/CSF-1R通路极可能是调节M2巨噬细胞浸润的重要通路。我们提出假说：解毒颗粒预防肝癌术后复发转移可能与其抑制M2巨噬细胞向癌旁组织迁移有关。.本课题拟建立大鼠肝癌切除术后模型、构建肝癌细胞与M2巨噬细胞共培养体系，通过免疫组化、PCR、Transwell、划痕试验等技术，明确解毒颗粒是否抑制M2巨噬细胞向癌旁组织迁移；进一步研究解毒颗粒是否通过调节CSF-1/CSF-1R通路影响M2巨噬细胞向癌旁组织迁移。

原发性肝癌是严重影响人类生活和生存的恶性肿瘤之一。手术切除是治疗早期肝癌的推荐方法之一，但肝癌术后的高复发率及低生存率均不能令人满意。肝癌是炎症相关性肿瘤，肿瘤相关巨噬细胞是肿瘤炎性微环境的重要组成部分，其中M2型巨噬细胞能够分泌大量的炎症因子，在介导慢性炎症和恶性肿瘤的进展之间发挥着重要的作用。研究表明，M2型巨噬细胞可通过多种途径促进肝癌的复发和转移。课题组前期的研究表明，低浓度解毒颗粒组的转移率明显低于对照组；在癌旁组织中，低浓度解毒颗粒组中CD68的表达明显低于对照组。M2型巨噬细胞的诱导增殖依赖CSF-1，因此推测，解毒颗粒预防肝癌术后复发转移可能与其影响CSF-1/CSF-1R通路进而抑制M2型巨噬细胞向癌旁组织迁移有关。为此，本课题通过体内及体外实验，探讨解毒颗粒是否通过调节CSF-1/CSF-1R通路抑制M2型巨噬细胞向癌旁组织迁移进而达到预防肝癌术后复发转移。研究结果表明：体内实验表明，解毒颗粒具有预防肝癌术后转移的作用；体内及体外实验表明，解毒颗粒可以影响CSF-1/CSF-1R通路进而抑制M2型巨噬细胞向癌旁组织迁移。以上研究结果为解毒颗粒在临床上的广泛应用提供与提供一定的理论支持。