白花丹醌调控HIF-1α/EMT抑制肝癌侵袭转移机制研究

Previous studies have found that Plumbagin inhibit the growth of hepatocellular carcinoma and inhibit tumor angiogenesis. Hepatocellular carcinoma is a highly invasive and highly metastatic tumor, in addition to the neovascularization provide a basic conditions to tumor invasion and metastasis, HIF-1a can regulate the process of epithelial cell stromal transformation and promote the occurrence of tumor invasion and metastasis. According to the basis of the previous study, this study proposes a scientific hypothesis that Plumbagin can inhibit the proliferation, migration and invasion of hepatocarcinoma cells by regulating the HIF-1a/EMT, Which can inhibit tumor metastasis. Establish three models including EMT model of Hepatocellular carcinoma in vitro, transplanted tumor model of Hepatocellular carcinoma in vivo, and the model of Hepatocellular carcinoma in vivo by Intravenous injection of SMCC7721-GFP. To study the expression of HIF-1α, which is closely related to the migration and invasion of hepatocarcinoma cells, and the expression of HIF-1α-mediated EMT-related marker protein by Western blot, immunofluorescence, immunohistochemistry, siRNA. To clarify the site of HIF-1a / EMT-related signaling pathway that affected by Plumbagin and the specific anti-hepatocarcinoma mechanism of Plumbagin in hypoxic environment. Providing more scientific basis for the development of Plumbagin. Guangxi Province is the main origin of plant Plumbago zeylanica L, its development and utilization will promote the economic and social development in Guangxi.

前期研究发现白花丹醌抑制肝癌移植瘤的生长，抑制肿瘤血管生成。肝癌是一种高侵袭、高转移性的肿瘤，除了新生血管生成为肿瘤的侵袭和转移提供了基础条件之外，HIF-1a能调控上皮细胞间质转化过程，促进肿瘤侵袭转移的发生。本项目拟在前期基础上提出了“白花丹醌通过调控HIF-1a/EMT干预肝癌细胞增殖、迁移、侵袭从而能抑制肿瘤转移”的科学假说，建立体外肝癌EMT模型和体内肝癌移植瘤模型、SMCC7721-GFP体内肝癌转移模型，通过western blot、免疫荧光、免疫组化、SiRNA等技术深入研究白花丹醌对与肝癌细胞侵袭转移密切相关的HIF-1α及EMT相关标志物的表达，弄清白花丹醌影响HIF-1a/EMT相关信号通路的部位，明确白花丹醌在乏氧环境下抗肝癌的具体作用机制，为开发白花丹醌抗肝癌提供更多的科学依据。广西是“猛老虎”白花丹主要产地，资源丰富，它的开发利用将促进广西经济和社会的发展。

肝癌是一种高侵袭、高转移性的肿瘤， HIF-1a 能调控 EMT 过程，使上皮细胞向间质细胞表型转化，促进肿瘤侵袭转移的发生。本项目通过体内研究证实白花丹醌可显著抑制裸鼠肝癌肝内转移和肺转移，体外研究证实白花丹醌抑制人肝癌细胞的增殖、侵袭，同时诱导细胞凋亡，其作用机制可能与下调HIF-1α蛋白及其下游靶基因蛋白如C-MYC、VEGF、TWIST、MMP9等表达有关，同时也发现白花丹醌有效抑制肝癌 SMMC-7721 细胞上皮-间质转化，白花丹醌作用使EMT相关标志物蛋白E-cadherin升高，并且N-cadherin, vimentin, Snail水平降低，使用通路抑制剂抑制PI3K/AKT/mTOR通路后，HIF-1α的表达下调，说明人肝癌细胞中HIF-1α 的表达可能与PI3K/AKT/mTOR通路相关。以上研究结果为白花丹醌的临床应用提供一定的科学参考。