藤龙补中汤对M2型肿瘤相关巨噬细胞分化及其介导大肠癌转移作用研究

Colorectal cancer is the third most frequently diagnosed malignancies worldwide. Metastasis remains the principal cause of treatment failure in patients with colorectal cancer. Inflammation has been recognized as the seventh hallmarks of the cancer, and contributes to colorectal cancer initiation, progression and metastasis. M2 type tumor-associated macrophages (TAM) are the major players of the cancer-related in?ammation. The polarization of TAM is regulated by JAK-STAT signal transduction. IL-10 and arginase I (Arg-1) are the main phenotypic genes of TAM. TAM can promote tumor metastasis which is associated with EGF and MMP9. TAM has been recognized as a novel target for prevention of metastasis. We have confirmed Teng-Long-Bu-Zhong-Tang (TLBZT), a herbal formula based on detoxification and spleen-tonifying, have significant anti-cancer effects. TLBZT may induce apoptosis and cell senescence in colorectal cancer cells. TLBZT also inhibit metastasis of colorectal cancer accompanied by TAM down-regulation in metastatic lesions, which suggested TAM may contribute to TLBZT elicted anti-metastasis effects. However, the cellular and molecular mechanism need further explore. The project will continue to explore the effects of TLBZT on TAM polarization and it mediated metastasis in colorectal cancer. To study the effects of TLBZT on IL-4 induced TAM polarization (marker, phenotypic and functional proteins). Based on JAK-SATA pathway to understand the molecular mechanism of the herbs on TAM polarization. In cell co-culture model, to observe the effects TLBZT on TAM mediated adhesion, invasion and migration in colorectal cancer cells. From the perspective of EGF and MMP-9 to elucidate the effective mechanism of the herbs on TAM mediated metastatic behaviors in colorectal cancer cells. And to study the effects of TLBZT against TAM associated colorectal cancer metastasis in vivo. The project will provide new insight into Traditional Chinese Medicine approaches for prevention and treatment of colorectal cancer metastasis.

大肠癌是全球第三大恶性肿瘤，转移是大肠癌治疗难点；M2型肿瘤相关巨噬细胞（Tumor-associated macrophage，TAM）与大肠癌转移密切相关，已成为大肠癌转移防治新的靶标；TAM活化与JAK-STAT信号转导相关，其介导肿瘤转移与EGF、MMP9相关。项目组前期研究表明：藤龙补中汤（专利号ZL200910197565.2）可促大肠癌细胞凋亡、细胞衰老；抑制大肠癌转移，降低TAM在转移灶的分布，其作用基础亟待研究。 项目拟基于标志蛋白、表型蛋白及功能蛋白研究藤龙补中汤对IL-4诱导TAM分化的影响，从JAK-STAT通路研究其信号转导；在细胞共培养体系中，研究藤龙补中汤对TAM介导大肠癌细胞粘附、移动及侵袭等转移生物学行为影响，从EGF、MMP9角度揭示其作用基础；动物模型中研究藤龙补中汤对TAM介导大肠癌转移的治疗作用；为中医药防治大肠癌转移提供新的科学基础。

大肠癌是全球最常见的恶性肿瘤之一，M2型肿瘤相关巨噬细胞巨噬细胞（TAMs）在大肠癌进展中发挥重要作用，参与大肠癌转移和血管生成。本课题组以解毒、利湿、健脾立法，研发了治疗大肠癌的中药复方藤龙补中汤，本研究在细胞与动物模型中研究了藤龙补中汤对TAMs分化及其介导大肠癌转移的作用。. 在细胞模型中，采用IL-4诱导TAMs分化。结果显示，藤龙补中汤可以抑制IL-4诱导的Arg-1和IL-10表达，提示藤龙补中汤可以抑制TAMs分化。藤龙补中汤可以抑制JAK3-STAT6磷酸化。藤龙补中汤还可降低TAMs表达和分泌VEGF-A。藤龙补中汤可以降低EGF表达，抑制TAMs介导的大肠癌细胞粘附，EGF可以拮抗藤龙补中汤的作用。藤龙补中汤可以抑制TAMs介导的大肠癌细胞移动。藤龙补中汤可以降低MMP-9表达，抑制TAMs介导的大肠癌细胞侵袭，MMP-9过表达可以拮抗藤龙补中汤的作用。. 动物模型研究中，采用尾静脉注射肿瘤细胞造模大肠癌肺转移，采用藤龙补中汤治疗。治疗结束后，取肺称重，计数肺表面转移结节。结果显示，藤龙补中汤治疗后，小鼠肺质量和肺表面转移结节显著降低，提示藤龙补中汤可以抑制大肠癌肺转移。免疫组化研究显示，藤龙补中汤可显著降低转移灶TAMs；藤龙补中汤还可下调VEGF-A表达，抑制大肠癌转移灶血管生成；提示TAMs参与藤龙补中汤对大肠癌肺转移的作用。. 本研究明确了藤龙补中汤对TAMs分化及其介导大肠癌转移的作用，初步阐明了藤龙补中汤作用的分子机制；丰富和完善了大肠癌中医治疗方法，深化了中医解毒、利湿、健脾治疗大肠癌的科学内涵，为中医药防治大肠癌转移提供了新的科学基础。