基于Ras信号通路研究延龄草皂苷成分抗结肠癌物作用机制

Colon cancer is a common malignant tumor. Trillium tschonoskii Maxim., one of four famous Traditional Chinese Medicine in Tu Jia Zu, has been used for hemostasis, analgesia, detumescence. In the course of our studies we found that the total saponins of T. tschonoskii inhibited proliferation of colon cancer cells in vitro and vivo; In lead of activity isolation from the total saponins, 9 saponin subatances isolated and identified, in which TTB1-3 were found to be the main activity constituents. TTB1-3 can react with RAS protein by computer simulation. Moreover, the total saponins can download RAF and AKT gene expression lever in vitro and vivo. In the present study, we will research resist activity and mechanism of TTB1-3 to RAS signal way in molecular, cell and animal levels on colon cancer cells, verifying that TTB1-3 can inhibit tumor cell growth and induce apoptosis via RAS signal way. The results of this project, will clarify the molecular mechanism of T. tschonoskii on colon cancer, as well as screening anti-tumor lead compounds from T. tschonoskii, which providing the basis for the research and development of new anticancer chemotherapy drugs.

结肠癌是消化系统中最常见的恶性肿瘤，延龄草是土家族四大名药之一，有消痛去肿、止血解毒等功效。本项目前期研究发现，延龄草总皂苷在体内外均有抗结肠癌活性；通过活性追踪分离总皂苷得到9个皂苷单体，其中TTB1、TTB2和TTB3能抑制结肠癌细胞增殖；计算机模拟筛选发现此3个化合物具有与癌基因产物RAS蛋白结合的潜在能力；RT-PCR显示总皂苷能抑制体外CT-26细胞和体内肿瘤组织中RAS通路下游RAF、AKT基因表达。提示延龄草活性成分可能通过抑制RAS信号传导通路而抑制肿瘤生长，发挥解毒作用。本研究以结肠癌细胞为研究对象，在分子、细胞和动物水平上分析TTB1-3对RAS通路的抑制作用及分子机制，验证TTB1-3能通过抑制RAS信号而抑制肿瘤细胞生长和诱导肿瘤细胞凋亡。本项目的成功实施，将阐明延龄草抗结肠癌药理活性的分子机制，解析延龄草解毒功效的现代科学内涵，也为新型抗癌化疗药物的研发奠定基础。

结肠癌是消化系统中最常见的恶性肿瘤，延龄草是土家族四大名药之一，有消痛去肿、止血解毒等功效。本项目前期研究发现，延龄草总皂苷在体内外均有显著的抗结肠癌活性；通过活性追踪分离总皂苷得到9种皂苷单体，其中TTB1、TTB2和TTB3能抑制结肠癌细胞增殖；计算机模拟筛选发现此3个化合物具有与癌基因产物RAS蛋白高亲和结合的潜在能力；RT-PCR显示总皂苷能抑制体外CT-26细胞和体内肿瘤组织中RAS通路下游RAF、AKT基因表达。提示延龄草活性成分可能通过抑制RAS信号传导通路而抑制肿瘤生长，发挥解毒作用。本研究以结肠癌细胞为研究对象，首先在分子和细胞水平分析TTB1-3对结肠癌细胞CT26的抑制作用及分子机制。结果表明，TTB1-3能显著抑制结肠癌细胞CT26增殖，抑制细胞凋亡，阻断细胞分裂周期，机制可能是TTB1-3作用于RAS蛋白，从而影响下游ERK/AKT蛋白功能。其次以结肠癌小鼠为模型，在动物水平上分析TTB1-3对结肠癌的作用及机制。结果表明验证TTB1-3能通过抑制RAS信号而抑制肿瘤细胞生长。本项目为延龄草临床应用及民族药物资源开发提供理论基础，同时为新型抗结肠癌药物的研发提供了依据。