14-3-3ζ/TGF-β/smad通路介导胃癌上皮-间质转化及七方胃痛颗粒干预的研究

Epithelial-mesenchymal transition(EMT) is closely related with gastric cancer’s invasion and metastasis, in which TGF-β/smad signaling pathway plays an important role. A number of researchs showed that overexpression of protein 14-3-3zeta(14-3-3ζ) in some carcinomas leads to an activation of TGF-β/smad signaling pathway and as a result EMT occurs while study reporting the relationship between 14-3-3ζ/TGF-β/smad pathway and gastric cancer’s EMT is few. Our team’s previous serial researchs have confirmed that Qifangweitong Granules can prevent and stop the formation of gastric cancer as it can effectively prevent and control the canceration of chronic atrophic gastritis and suppress gastric cancer from growing. However, it needs to do the further research for whether Qifangweitong Granules can stop the invasion and metastasis of gastric cancer through inhibiting its EMT or not. We suppose that 14-3-3ζ could induce gastric cancer’s EMT by regulating TGF-β/smad signaling pathway and Qifangweitong Granules could suppress gastric cancer’s EMT by inactivating 14-3-3ζ/TGF-β/smad pathway. Experimental techniques include RT-PCR, Western Blot, immunohistochemistry, immunofluorescence, laser confocal microscopy, lentivirus transfection, and assays like MTT, wound healing assay, transwell migration assay will be applied to testify the hypotheses above through the experiment on rats which have been built up the gastric cancer model and the experiment in vitro on human gastric adenocarcinoma cell’s EMT, all of which is available for discovering the truth of Chinese Medicine in controlling gastric cancer.

上皮-间质转化（EMT）与胃癌侵袭转移密切相关，TGF-β/smad通路在此起着重要作用，部分肿瘤中14-3-3ζ蛋白过表达可活化该通路并促发EMT，但14-3-3ζ/TGF-β/smad通路与胃癌EMT的相关性研究报道甚少。我们前期课题的系列研究证实七方胃痛颗粒可防治胃癌形成：该方药对慢性萎缩性胃炎癌前病变及胃癌有显著防治作用，但它能否通过抑制胃癌EMT来阻止胃癌发生侵袭转移目前尚待进一步研究。我们推测14-3-3ζ可通过调控TGF-β/smad通路介导胃癌EMT，七方胃痛颗粒可能通过调节14-3-3ζ/TGF-β/smad通路来抑制胃癌EMT。本项目拟通过对大鼠胃癌的动物试验和胃癌细胞EMT的体外试验，运用RT-PCR、Western Blot、免疫组化、免疫荧光及激光共聚焦显微镜、慢病毒转染等技术及MTT、细胞划痕试验、细胞侵袭试验等试验来验证如上假说，为中医药防治胃癌提供客观依据。

背景：胃癌为起源于上皮的恶性肿瘤，是世界上第四高发的肿瘤，致死率位列第二。胃癌是个涉及多基因、多信号的复杂过程，是一个非线性复杂系统，其中胃癌上皮-间质转化是重要因素。浸润性侵袭转移是导致肿瘤发生发展的重要原因。.方向：拟通过体内及体外实验，采用现代分子生物技术，从基因、蛋白等分子水平层面观察七方胃痛颗粒对胃癌14-3-3ζ/TGF-β/smad通路以及对胃癌EMT和获得侵袭转移能力的干预作用。探讨胃癌侵袭转移形成过程中的病理生理机制，以及探讨14-3-3ζ/TGF-β/smad通路介导胃癌EMT并使其获得浸润侵袭转移能力的作用和机制，为中医药防治胃癌提供客观依据。.内容：拟从胃癌上皮-间质转化的角度，通过建立大鼠胃癌模型，采集大鼠胃癌组织及血液的体内实验和胃腺癌细胞EMT的体外试验，运用RT-PCR、Western Blot、免疫组化、免疫荧光及激光共聚焦显微镜技术、慢病毒转染等生物技术及MTT、细胞划痕试验、细胞侵袭试验等试验，探讨14-3-3ζ/TGF-β/smad通路介导胃癌EMT和获得浸润侵袭转移能力过程中的作用及机制，为临床通过14-3-3ζ/TGF-β/smad通路早期干预EMT治疗胃癌提供理论依据及潜在干预指标；并探讨七方胃痛颗粒对胃癌EMT的干预作用和机制。 .科学意义：本课题从基因转录和蛋白结合位点水平探讨七方胃痛颗粒对胃癌中14-3-3ζ/TGF-β/smad通路对胃癌EMT的调控作用以及对14-3-3ζ在胃癌中与EMT的关系及其机制与TGF-β/smad信号通路的关系进行探讨。然而目前尚未有用复方中药调控胃癌中14-3-3ζ/TGF-β/smad通路的研究。