慢性应激激活β2-AR调控β-catenin核转位促肝癌侵袭转移的分子机制研究

The metastasis and recurrence are the main cause of death in hepatocellular carcinoma（HCC）. Many studies reveal that chronic psychological stress promotes the progression of malignant tumors by activating the sympathetic nervous system（SNS）. It’s identified by our previous study that sustained activation of SNS enhanced hepatocarcinogenesis in cirrhotic liver. Our recent study found that the level of chronic stress in HCC patients was related to the prognosis, and chronic stress promoted the invasion and metastasis of HCC by activating the SNS. The results of PCR-arrays suggested that chronic stress could regulate the activation of β-catenin to promote the downstream gene expression by activating β2-AR of HCC cells. Therefore, the hypothesis is introduced that chronic stress facilitates the invasion and metastasis of HCC through regulating β-catenin nuclear translocation by activating β2-AR. Firstly, the heart rate variability (HRV) of HCC patients would be detected to assess the level of chronic stress of patients. The specific mechanism that chronic stress promoted the invasion and metastasis of HCC by activating of β2-AR would be confirmed. And the molecular mechanism of β-catenin nuclear translocation after activation of β2-AR would be investigated used some high-tech technique (CRISPR-Cas9, TOP/FOP-flash).The aim of this study is to reveal the new theory of the invasion and metastasis regulated by psychological stress，and to provide a new theoretical and experimental basis for prevention the metastasis and recurrence of HCC.

肝癌的转移和复发是患者死亡的主要原因。研究表明：慢性心理应激能激活交感神经系统促进恶性肿瘤的进展。本课题组之前的研究证实：肝硬化基础上，肝脏交感神经的持续兴奋能促进肝癌发生。我们近期研究发现：患者慢性应激水平与肝癌预后密切相关，慢性应激激活交感神经系统促进肝癌侵袭转移；芯片结果提示其机制可能与激活β2-AR调控β-catenin入核促下游基因表达有关。因此我们提出“慢性应激激活肝癌细胞β2-AR调控β-catenin核转位促肝癌侵袭转移”的假说。本项目拟利用心率变异检测等技术评估患者慢性应激状态，证实慢性应激通过激活β2-AR促肝癌侵袭转移，再利用CRISPR-Cas9,TOP/FOP-flash等技术研究激活β2-AR如何调控β-catenin核转位，促下游基因表达调控肝癌侵袭转移的分子机制，揭示心理应激调控肝癌侵袭转移的新理论，为临床从心理应激角度防治肝癌转移复发提供新的理论依据。

肝癌是最常见的恶性肿瘤之一。恶性肿瘤患者在诊疗过程中多处于长期的慢性心理应激状态，慢性应激被证实通过调控肿瘤细胞，肿瘤微环境和机体免疫系统等影响肿瘤的进展。我们前期研究发现慢性心理应激能促进肝癌的进展，且可能与激动β2-AR调控β-catenin通路活化促进肝癌细胞侵袭转移有关。本课题旨在研究慢性应激激活β2-AR调控β-catenin核内转位促肝癌侵袭转移的分子机制，为从心理应激角度防治肝癌的复发转移提供新的理论基础和实验依据。.本项目利用HRV指标及应激激素检测联合评价肝癌患者慢性应激水平，研究证实慢性应激高水平的患者预后较差；基于慢性应激动物模型，着重研究慢性应激激活交感神经系统对肝癌转移的调控作用；通过PCR芯片、TOP/FOP-flash等技术研究阐明激动β2-AR后抑制β-catenin泛素化降解，促进β-catenin/Vimentin信号通路活化，促肝癌细胞EMT进而调控肝癌转移的分子机制。通过本课题的研究进一步证实慢性应激调控交感神经系统促进肝癌转移和复发；β2-AR是介导慢性应激促肝癌转移的关键受体；慢性应激通过激活交感神经系统调控β2-AR/β-catenin/Vimentin信号通路活化，促进肝癌细胞EMT进而促肝癌转移。阐明慢性应激调控肝癌转移的分子机制，将可能为临床上预测肝癌患者预后，以及探索防治肝癌复发的治疗策略提供潜在靶点。