隐丹参酮调控FoxO1抑制非小细胞肺癌增殖的机制研究

Non-small cell lung cancer (NSCLC) is the most common respiratory malignancy, its morbidity and mortality are the first of all malignant tumors, causing great harm to people's life and health, there is still a lack of effective treatment. The literature shows that the cryptotanshinone has a good anti - tumor prospect. Our results showed that cryptotanshin could induce the apoptosis of A549 cells in a dose-dependent manner, inhibit the proliferation of A549 cells and block the cell cycle in G1 phase. After treatment with cryptotanshinone, FoxO1 was transfected into the nucleus of A549 cells, and the expression level of p27kip was increased, and the level of p-FoxO1 significantly decreased but the total FoxO1 protein was not significantly changed. Whether the cryptotanshinone can inhibit the tumor by regulating FoxO?what is the specific molecular mechanism? It is suggested that the phosphorylation of Thr24 at FoxO1 may play an important role in it. In this study, the molecular mechanism of cryptotanshinone-controlled FoxO1 was studied by using the method of transfection of FoxO1 SiRNA and FoxO1 plasmid with Thr24 phosphorylation site deletion, western blot, flow cytometry and immunofluorescence to detect the effect of cryptotanshinone on the expression of FoxO1 itself and its upstream and downstream signaling molecules in order to further explore the molecular mechanism of FoxO1 in inhibiting the proliferation of non-small cell lung cancer . In situ cancer model by HE staining pathological analysis, immunohistochemistry and other methods to verify the hypothesis, to find non-small cell lung cancer drug target research and development to provide new ideas. This project is expected to find a new target for the anti-tumor effect of cryptotanshinone and provide a more scientific basis for the development of new methods for the treatment of non-small cell lung cancer.

非小细胞肺癌（NSCLC）是最常见的呼吸系统恶性肿瘤，对人们生命健康造成极大威胁，目前尚缺乏有效的治疗方法。文献表明中药单体隐丹参酮具有良好的抗肿瘤前景，我们前期实验表明隐丹参酮可剂量依赖性诱导NSCLC A549细胞凋亡、抑制增殖及将细胞周期阻滞于G1期，同时隐丹参酮作用下FoxO1转位入核明显，FoxO1靶蛋白p27kip表达升高，同时胞内p-FoxO1水平显著降低，但FoxO1总蛋白无明显变化。文献表明FoxO1的Thr24位点磷酸化可能在其中起到重要作用。本项目拟采用SiRNA转染、过表达Thr24磷酸化位点缺失突变的FoxO1质粒转染、western blot、流式细胞术、免疫荧光等方法，检测隐丹参酮对FoxO1本身及上下游信号分子的影响，同时在裸鼠原位癌模型中通过HE染色病理切片分析、免疫组化等方法验证该假说，以期深入探讨隐丹参酮调控FoxO1抑制非小细胞肺癌增殖的分子机制。

非小细胞肺癌（NSCLC）是最常见的呼吸系统恶性肿瘤，对人们生命健康造成极大威胁，目前尚缺乏有效的治疗方法。中药单体隐丹参酮具有良好的抗肿瘤作用，本研究按计划开展以非小细胞肺癌细胞株为主要研究对象，结合动物实验，利用Western blot、流式细胞术、激光共聚焦、SiRNA、过表达质粒转染等技术，深入探讨FoxO1在隐丹参酮抑制非小细胞肺癌增殖中的作用，并检测隐丹参酮对FoxO1本身及其上下游信号分子表达的影响，以期深入阐明隐丹参酮抗肿瘤的作用机制。实验表明，隐丹参酮可剂量依赖的减少NSCLC裸鼠肿瘤体积和重量、抑制肿瘤细胞增殖和促进肿瘤细胞凋亡；隐丹参酮抑制NSCLC细胞增殖，并增加G1期的肿瘤细胞数量；FoxO1在NSCLC 细胞中位于细胞浆内，隐丹参酮处理可引起FoxO1由胞浆向胞核内转位；隐丹参酮处理A549细胞，FoxO1总蛋白无明显变化，但p-FoxO1水平显著降低，靶蛋白p27kip表达水平升高。表明隐丹参酮抑制非小细胞肺癌增殖机制与抑制FoxO1磷酸化及促进FoxO1入核部分相关。