交感神经调控kupffer细胞ATF2活化介导其替代激活促肝癌侵袭转移的分子机制研究

The invasion and metastasis are the main cause of death in hepatocellular carcinoma(HCC). Many studies reveal that the regulation of sympathetic nervous system plays an important role for the initiation and progression of cancer. It’s identified by our last NSFC project that activation of adrenergic receptors by sympathetic nervous system enhanced hepatocarcinogenesis in cirrhotic liver by production of cytokines from activated kupffer cells. Other reports showed that the invasion and metastasis were enhanced by alternative activation of kupffer cells by adrenergic receptors, and mediated with activated translation factors(ATF/CREB etc.). Therefore, the hypothesis is introduced that the invasion and metastasis in HCC is facilitated by cytokines from alternative activated kupffer cells regulated by sympathetic nervous system through activation of ATF2. Firstly, the specific subset of adrenergic receptors for alternative activation of kupffer cells would be identified. And, the molecular mechanism of production of cytokines through alternative activation of kupffer cells mediated by activation of ATF2 would be also investigated used some high-tech technique(CRISPR-Cas9, CHIP and base substitution) . The aim is to establish the new theory of the invasion and metastasis regulated by alternative activated kupffer cells by regulation of sympathetic nervous system, and to find the new molecule targets for the therapy in HCC.

侵袭转移是导致肝癌预后较差的主要原因。研究发现交感神经对肿瘤的发生发展具有重要的调控作用。我们的上一个基金项目已证实交感神经激动肾上腺素能受体（ARs）能够介导kupffer细胞的活化表达多种细胞因子促进肝硬化肝脏癌变的发生；另有报道亦发现ARs激动能诱导巨噬细胞替代激活促进肿瘤侵袭转移，且与ATF/CREB等转录因子参与有关。因此我们提出“交感神经调控kupffer细胞ATF2活化介导其替代激活表达促癌细胞因子促进肝癌侵袭转移”的假说。本项目拟首先鉴定交感神经调控kupffer细胞替代激活的关键ARs亚型，再利用CRISPR-Cas9、ChIP和点突变等新技术研究kupffer细胞替代激活过程中ATF2活化介导关键细胞因子转录调控，进而促肝癌侵袭转移的分子机制。揭示kupffer细胞替代激活促肝癌侵袭转移的交感神经调控新理论，发现潜在的干预新靶点，为临床防治肝癌的侵袭转移提供理论依据。

近年来的研究表明，肝脏的交感神经参与了肝脏的各种生理和病理生理的过程。我们的前期研究，已证实交感神经调控与癌症发生、发展和侵袭转移具有明显的相关性。本项目旨在阐明交感神经通过调控kupffer细胞 ATF2活化介导多种促癌细胞因子转录调控，进而促进肝癌侵袭转移的分子机制。.本项目主要利用肝癌动物模型，CRISPR-Cas9、ChIP和点突变等新技术鉴定交感神经调控kupffer细胞替代激活的关键的肾上腺素能亚型；研究临床病例和动物模型中交感神经调控kupffer细胞替代激活和ATF2表达与肝癌侵袭转移的相关性；研究肾上腺素能受体亚型活化对kupffer细胞替代激活的调控作用和机制及其对肝癌细胞侵袭转移的影响；研究交感神经调控kupffer细胞替代激活过程中ATF2活化介导关键细胞因子转录调控的分子机制。.通过本课题的研究发现临床肝癌标本中交感神经纤维密度和交感神经递质浓度增高与患者术后生存时间呈负相关，证实交感神经系统的兴奋能够促进肝癌原位移植瘤模型中肿瘤的生长和远处转移。交感神经系统的兴奋能够促进肝癌原位移植瘤模型中肿瘤的血管侵犯和远处转移，同时发现交感神经递质能够促进体外培养的原代 kupffer细胞激活，释放多种细胞因子；进一步研究发现，交感神经兴奋后释放的异丙肾上腺素等神经递质能够通过激活β2肾上腺素能受体促进kupffer细胞的活化，调控炎症因子的释放进而促进肝癌的发生。本项目揭示肿瘤微环境中kupffer细胞替代激活促肝癌侵袭转移的神经调控新理论，发现潜在的干预新靶点，为临床防治肝癌的侵袭转移提供新的理论基础和实验依据。.