神经心理应激对非小细胞肺癌CREB信号传导通路作用机制研究

Neuropsychological stress has been associated with increased cancer incidence rate and mortality. Some in vivo studies showed that neurotransmitter responses to neuropsychological stress and its resulting hyperactive cAMP response element binding protein (CREB) may significantly promote the progression of cancer. Our preliminary results demonstrated that CREB and its regulated signaling pathway was highly expressed and hyperactivated in NSCLC and correlated with tumor clinicopathological findings and prognosis. However, the underlying mechanisms of neuropsychological stress on the CREB and its regulated signal transduction pathway in NSCLC have never been reported in the worldwide. In this study, using an established mouse model of neuropsychological stress, we will investigate the effects of neuropsychological stress on neurotransmitters with their receptors and tumor proliferation, invasion, migration and apoptosis in nude mice carrying xenografts from NSCLC cell lines. Meanwhile, we will further evaluate the changes of DNA, mRNA and protein levels of CREB and its associated signaling proteins in the nude mice, and then establish a CREB signaling network regulated by neuropsychological stress. The practical results of this work could provide new thoughts of molecular mechanisms of neuropsychological stress on the CREB and its regulated signal transduction pathway in NSCLC and establish theoretical basis of optimal clinical management and preventive strategies of NSCLC patients.

目前发现神经心理应激（NPS）使癌症发病率和死亡率显著上升，体外研究表明NPS促使某些肿瘤相关神经递质及其受体表达增高、激活c-AMP反应元件结合蛋白（CREB），从而促进肿瘤发生、发展。本课题组前期研究中亦发现，CREB及其信号传导通路中某些信号因子在非小细胞肺癌（NSCLC）中表达增高并与临床病理及预后关系密切；而有关NPS对CREB信号传导通路在NSCLC中的调节机制国内外还未见报道。本课题拟利用裸鼠荷NSCLC模型，研究NPS对荷瘤鼠血清和肿瘤组织相关神经递质及受体表达的影响，以及对肿瘤生长、细胞凋亡及侵润、转移相关蛋白表达的影响，并在基因、mRNA和蛋白水平研究CREB信号传导通路关键信号因子在NPS作用下的表达变化，筛选并构建NPS和CREB信号传导通路的信号传导网络，明确NSCLC中NPS对CREB信号传导通路的作用机制，为探讨新的NSCLC治疗和预防策略提供理论依据。

本课题为探讨神经心理应激（NPS）因素对非小细胞肺癌（NSCLC）肿瘤细胞的影响及其机制，建立了NPS裸鼠荷NSCLC腺癌细胞株动物模型，通过酶联免疫吸附试验（ELISA）技术检测荷瘤鼠血清中肾上腺素、去甲肾上腺素和可的松等相关神经递质的表达，以及对荷瘤鼠肿瘤生长指标（体积和重量）和细胞凋亡的检测，证明NPS确能明显促进肿瘤生长，亦表明NPS荷NSCLC腺癌细胞株动物模型建立成功；基于此模型，进一步采用免疫组化方法检测了动物肿瘤组织中CREB和p-CREB的表达状况，结果表明NPS及其相关神经递质可引起与之密切相关的CREB传导通路中CREB和p-CREB的表达明显增高；本研究还采用了ELISA和western blotting方法分别在NSCLC患者血清和肿瘤组织中研究了CREB和p-CREB的表达状况，结果表明p-CREB在NSCLC患者血清和肿瘤组织中的表达明显高于正常人血清和NSCLC患者瘤旁组织中的表达。此项研究揭示了NPS对NSCLC发生、发展在CREB信号传导通路中的部分机制，将来进一步的深入研究有助于发现NPS及CREB相关信号传导通路在NSCLC发生发展机制中的作用，为今后预防、筛查、治疗NSCLC提供新的途径。