消癥化瘀法调控Wnt通路关键蛋白β-catenin和cyclinD1影响肺癌干细胞增殖的研究

The lung cancer is a malignant tumor which threat our life badly. Surgical operation and radiotherapy can not achieve good results. Its relapse and metastasis is a difficult question in treatment. Dissolving lumps and removing blood stasis have a good curative effect in preventing the relapse and metastasis of lung cancer. Our earlier studies discover that the HuiSheng-liqud which represent dissolving lumps and removing blood stasis , can restrain lung cancer cells. On these bases , we will use the HuiSheng-liqud act on the lung cancer stem cells which chosen by FCM, then detect the gene expression and protein expression of β-catenin, Cyclin D1 and OCT-4 in lung cancer stem cells through Western Blot and RT-PCR, and research the lung cancer tissue in nude mice. With all of these results，verify the relationship between the HuiSheng-liqud and the lung cancer stem cells. Then explore the mechanism if the dissolving lumps and removing blood stasis could influence the cell proliferation of lung cancer stem cells, then restrain proliferation of lung cancer cells. The research results could verify target sites which the dissolving lumps and removing blood stasis treat lung cancer cells on, enrich its theoretical frame, provide scientific evidence in finding better therapeutic plans. In a word , the science value and clinical value of this project are very important.

肺癌是威胁人类的高发恶性肿瘤，病死率高，现有手术加放、化疗治疗效果差，易复发转移。肺癌组织内存在肺癌干细胞，其增殖是肺癌细胞增生的根源。消癥化瘀法是治疗肺癌的有效治法，本课题前期临床及动物实验研究显示，消癥化瘀法代表方回生口服液对肺癌细胞有抑制作用，在此基础上，以肺癌干细胞为研究对象，自Wnt信号通路切入，应用Western Blot、RT-PCR等技术，研究消癥化瘀复方回生口服液干预后的肺癌干细胞在裸鼠体内的成瘤情况及β-catenin、cyclinD1、OCT-4表达的变化，探讨消癥化瘀法是否通过调控Wnt信号通路关键蛋白β-catenin和cyclinD1及肺癌干细胞核心转录因子OCT-4影响肺癌干细胞增殖、从而发挥抑制肺癌细胞生长的作用机制。研究结果有助于明确消癥化瘀法治疗肺癌的作用环节和靶点，丰富该治疗方法的现代理论内涵，为临床治疗方案的优化提供佐证，具有重要理论和临床实用价值。

肺癌是威胁人类的高发恶性肿瘤，病死率高，现有手术加放、化疗治疗效果差，易复发转移。肺癌组织内存在肺癌干细胞，其增殖是肺癌细胞增生的根源。消癥化瘀法是治疗肺癌的有效疗法，本课题前期临床及动物实验研究显示，消癥化瘀法代表方回生口服液对肺癌细胞有抑制作用，在此基础上，以肺癌干细胞为研究对象，自Wnt信号通路切入，应用Western Blot、RT-PCR等技术，研究消癥化瘀复方回生口服液干预后的肺癌干细胞在裸鼠体内的成瘤情况及β-catenin、cyclinD1、OCT-4表达的变化，探讨消癥化瘀法是否通过调控Wnt信号通路关键蛋白β-catenin和cyclinD1及肺癌干细胞核心转录因子OCT-4影响肺癌干细胞增殖、从而发挥抑制肺癌细胞生长的作用机制。研究结果有助于明确消癥化瘀法治疗肺癌的作用环节和靶点，丰富该治疗方法的现代理论内涵，为临床治疗方案的优化提供佐证，具有重要理论和临床实用价值。.消癥化瘀法代表方药回生口服液高、中剂量组和顺铂组成瘤的瘤重明显降低，抑瘤率明显升高；回生口服液高、中剂量可下调肺癌干细胞β-catenin、CyclinD1 及OCT-4 mRNA 及蛋白表达，从而推论，消癥化瘀法治疗肺癌的分子作用机制之一是通过抑制β-catenin、CyclinD1 及OCT-4的表达，从而抑制肺癌干细胞wnt信号转导通路异常激活及肺癌干细胞的活跃增殖。整个实验的研究结果也论证了项目组之前提出的假说。.科学意义：.项目研究内容以全新的视角，自肺癌干细胞水平深入，首次将传统治疗肺癌的常用疗法——消癥瘀化瘀法与肺癌干细胞的重要信号转导途径Wnt通路及其关键调控蛋白结合起来，建立以肿瘤干细胞为治疗靶点的中医药抗肿瘤研究的新思路、新途径。.实验结果可为临床研发中医药治疗肿瘤新方案及进一步开发抗肿瘤新型中药提供实验依据。