基于健脾活血解毒法的复方肠泰对大肠癌细胞自噬通路的干预作用及其与凋亡关系的研究

Temper weak, damp-heat is pathogenesis of colorectal cancer, Jianpi Huoxue detoxification is an important rule in the clinical treatment of colorectal cancer. Based on the theory, intestinal Jianpi Huoxue detoxification is the formulation of the task group concluded Thai created inside the courtyard, with good clinical curative effect, our preliminary study showed that the medicated serum can inhibit the proliferation of colon cancer cells in vitro, and induce the occurrence of autophagy and apoptosis, but the relationship bettwen autophagy and apoptosis in colon cancer cells remains unkown. This project aims to use immunofluorescence, RNAi, and fluorescence quantitative PCR to further explore the effect of Changtai on the signal pathway such as PI3K-Akt-mTOR, Beclin-1-PI3K- Ⅲ( hvps34 ); use 3-MA, autophagy inhibitor, to investigate whether the in vivo antitumor effect of Changtai dependent on autophagy pathway; and to explore the relationship between autophagy and apoptosis. This project will provide experimental evidence for the status and role of autophagy in Chinese medicine in a hot fields in tumor research.

脾气虚弱, 湿热瘀毒是大肠癌的病机要点，健脾活血解毒是临床治疗大肠癌的重要法则。基于健脾活血解毒法的复方肠泰是本课题组总结创制的院内制剂，临床使用20多年，具有良好疗效，我们的前期研究发现该方剂能在体外抑制大肠癌细胞增殖，并诱导其发生自噬与凋亡，但自噬在复方肠泰诱导大肠癌细胞凋亡中的作用及其机制尚不明确。本项目拟采用信号阻滞、免疫荧光、RNAi、荧光定量PCR、建立大肠癌裸鼠移植瘤模型等技术手段，进一步探讨复方肠泰对PI3K-Akt-mTOR、Beclin-1-PI3K-Ⅲ（hvps34）自噬通路及其调控因子的干预作用；利用3-MA阻断自噬后明确复方肠泰体内抑瘤作用是否依赖于自噬途径；重点阐述自噬与凋亡之间的内在关系。以期从诱导细胞自噬、自噬性死亡角度揭示健脾活血解毒法抗肿瘤作用机制和组方配伍的科学原理，并为中药在自噬这一肿瘤研究热点领域的地位和作用提供实验研究依据。

脾气虚弱, 湿热瘀毒是大肠癌的病机要点,健脾活血解毒是临床治疗大肠癌的重要法则。基于健脾活血解毒法的复方肠泰是本课题组总结创制的院内制剂,临床使用 20 多年,具有良好疗效,我们的前期研究发现该方剂能在体外抑制大肠癌细胞增殖,并诱导其发生自噬与凋亡,但自噬在复方肠泰诱导大肠癌细胞凋亡中的作用及其机制尚不明确。本项目采用信号阻滞、免疫荧光、RNAi、荧光定量 PCR、建立大肠癌裸鼠移植瘤模型等技术手段,通过复方肠泰对自噬和凋亡信号网络干预，从分子水平揭示复方肠泰抗癌的分子机制；在细胞水平明确自噬在复方肠泰诱导大肠癌细胞凋亡中的调节作用；建立大肠癌移植瘤裸鼠模型，整体上考查复方肠泰体内抑瘤效应是否依赖于自噬途径。从诱导细胞自噬、自噬性死亡角度揭示健脾活血解毒法抗肿瘤作用机制和组方配伍的科学原理,并为中药在自噬这一肿瘤研究热点领域的地位和作用提供实验研究依据。