基于“循环肿瘤细胞”探讨蟾灵膏对HIF-1α/TGF-β介导的EMT效应在结直肠癌肝转移中的作用机制

Colorectal liver metastasis（CRLM）is the most important cause of death, and the mechanism is not yet clear. Circulating tumor cells (CTCs) are the required process of CRLM, which is closely related to the epithelial mesenchymal transition (EMT) of the tumor cells. Studies have shown that HIF-1 alpha and TGF- beta can mediate EMT to play an important regulatory role in the metastasis of many tumors. However, the role of EMT mediated by HIF-1alpha/TGF-beta in CRLM has not been clearly explained. blocking the occurrence of cancer cells in EMT may be the key to the treatment of CRLM, is in line with Chinese medicine "prevention before disease" "disease prevention" thought. Previous studies have shown that Chan ling gao can inhibit the liver metastasis of LoVo in colorectal cancer, and can reduce the levels of HIF-1 alpha, MMP-2, VEGF and other proteins. Therefore, we put forward the hypothesis that HIF-1 alpha /TGF- beta mediated EMT may be a potential target of CRLM. Chan ling gao may reduce the occurrence of CRLM by regulating the formation of CTCs by regulating EMT. This project will be further discussed based on early Chan ling gao intervention HIF-1 alpha/TGF-beta mediated EMT in SW480 liver metastasis of colorectal cancer mechanism.alpha /TGF- beta may be a potential target of CRLM. Chan Ling Gao may reduce the occurrence of CRLM by regulating the formation of CTCs by regulating EMT. Based on our previous work, we will further explore the mechanism of the effect of Chan Ling Gao on interfering the EMT effect mediated by HIF-1 alpha /TGF- beta in SW480 liver metastasis of colorectal cancer.

结直肠癌肝转移（CRLM）是导致患者死亡的最主要原因，目前机制尚未清楚。循环肿瘤细胞（CTCs）是CRLM的必经过程，其发生与瘤细胞上皮间质转化（EMT）密切相关。研究表明，HIF-1α和TGF-β均可介导EMT在多种肿瘤的转移中发挥重要调节作用，而HIF-1α/TGF-β协同介导的EMT在CRLM中的作用尚未得到明确解释。阻断癌细胞EMT的发生可能是治疗CRLM的关键，正契合中医“未病先防”“既病防变”的思想。前期研究表明，蟾灵膏有抑制结直肠癌LoVo肝转移的作用，可下调HIF-1α、MMP-2、VEGF等蛋白的水平。因此我们提出假设：HIF-1α/TGF-β介导的EMT可能是CRLM的潜在靶点，蟾灵膏可能通过调节EMT影响CTCs的形成，从而降低CRLM发生。本项目将在前期基础上进一步探讨蟾灵膏干预HIF-1α/TGF-β介导的EMT在结直肠癌SW480肝转移中的作用机制。

结直肠癌肝转移（CRLM）是导致患者死亡的最主要原因。循环肿瘤细胞是CRLM的必经过程，其发生与瘤细胞上皮间质转化（EMT）密切相关，阻断癌细胞EMT的发生可能是治疗CRLM的关键。蟾灵膏是国医大师刘尚义教授五十余年临床经验方，常用于肿瘤患者的防治。前期研究表明，蟾灵膏有抑制结直肠癌LoVo细胞肝转移的作用，下调HIF-1α、MMP-2、VEGF等蛋白的水平，但蟾灵膏是否可通过HIF-1α/TGF-β调控EMT效应抑制结直肠癌的进展不得而知。基于此，本项目主要研究探讨了蟾灵膏通过EMT效应对结直肠癌的作用机制，通过4年的努力，主要完成以下工作：（1）明确了蟾灵膏能抑制人结直肠癌SW480细胞的增殖、迁移及侵袭；（2）蟾灵膏可上调E-cadherin蛋白表达，增加细胞-细胞间的粘附，降低上皮细胞游离性，下调MMP-9的分泌，降低胞外降解，抑制间质-上皮转化，降低间质细胞的侵袭性；（3）蟾灵膏可改善结直肠癌模型鼠的体质量，降低模型鼠肝脏重量及肝脏指数；抑制原位移植瘤的增长，减轻模型鼠的肿瘤负荷；（4）蟾灵膏有抑制斑马鱼结直肠癌模型肿瘤及血管的作用；（5）蟾灵膏可增强移植瘤组织中E-cadherin的含量，抑制HIF-1α、TGF-β1、MMP-9、Snail、VEGF的表达；增加转移瘤组织中E-cadherin、ZO-1的含量，抑制HIF-1α、TGF-β1、MMP-9、Snail的表达；（6）发表相关SC论文3篇；（7）协助培养研究生2名，其中1名获校级优秀硕士论文；（8）相关研究在国际论坛作报告2次，国内论坛作报告1次。该研究对蟾灵膏的临床推广应用提供了一定的支撑依据，促进了中医药事业的发展及人才培养。