Fas-FasL/TFF3-NF-κB介导胃癌细胞失巢凋亡与增殖机制及加味七方胃痛颗粒干预的研究

Our previous study had confirmed that Qifangweitong Granules exert a protective role to gastric mucosa and prevent the canceration of chronic atrophic gastritis, but its target is unknown. We consider the canceration of chronic atrophic gastritis refer to complex process involving multiple genes and multiple signals. Especially, the imbalance of proliferation and apoptosis is an important factor. Fas binding to its ligand FasL could induce anoikis, however, Trefoil factor 3(TFF3) could prevent cell anoikis through active NF-κB signaling, and up-regulating the transcription of FLIP lead to cell proliferation. In this study, we focus on the perspective of cell anoikis and proliferation, a nude mice metastasis model of human gastric cancer was established, collecting nude mouse gastric cancer tissues and blood in vivo and gastric cancer cells cultured in vitro experiments. Biological techniques such as RT-PCR, MTT, ELISA, Western Blot and RNA interference technology will use to explore the imbalance between Fas-FasL-FADD-Caspase-8 pathway-mediated downstream apoptosis and TFF3-NF-κB-ERK to promote cell proliferation on the formation of gastric cancer pathophysiological mechanisms, and explore the the effect of Qifangweitong Granules on cell anoikis and proliferation in intestinal type gastric cancer, which had demonstrated the clinical efficacy, further provide objective evidence for Chinese medicine in prevent canceration.

前期证实加味七方胃痛颗粒具有对胃粘膜的保护及对慢性萎缩性胃炎癌变的防治效应，但其靶点不明。项目组认为萎缩性胃炎癌变是个涉及多基因、多信号的复杂过程，其中增殖与凋亡的失衡是重要因素。Fas与配体FasL结合可诱导细胞失巢凋亡，TFF3通过激活NF-κB信号途径阻断细胞失巢凋亡并通过上调FLIP的转录而诱导细胞增殖。因此本课题从细胞失巢凋亡与细胞增殖的角度，通过建立裸鼠胃癌原位移植模型，采集裸鼠胃癌组织及血液的体内实验与胃癌细胞培养的体外实验，运用生物学技术RT-PCR、MTT、ELISA、Western Blot及RNA干扰等技术探讨Fas-FasL-FADD-Caspase-8通路介导下游细胞凋亡，与TFF3-NF-κB- ERK促进细胞增殖之间的失衡在肠型胃癌形成过程中的病理生理机制，并探讨加味七方胃痛颗粒对胃癌失巢凋亡与增殖的调节效应，为中医药防治胃癌提供客依据。

背景：胃癌是世界范围内最常见的恶性肿瘤之一。萎缩性胃炎癌变是个涉及多基因、多信号的复杂过程，是一个非线性复杂系统，其中增殖与凋亡的失衡是重要因素。细胞凋亡与增殖之间的失衡是导致肿瘤发生发展的重要原因。.方向：拟从基因水平层面观察加味七方胃痛颗粒中药对正常SGC7901细胞株、NF-κB基因沉默株、胃癌模型Fas-FasL-FADD-Caspase-8通路与TFF3-NF-κB- ERK通路以及这两条通路之间的影响，探讨胃癌形成过程中的病理生理机制，以及探讨七方胃痛颗粒对胃癌失巢凋亡与增殖的调节效应，为中医药防治胃癌提供客观依据。.内容：从细胞失巢凋亡与细胞增殖的角度，通过建立裸鼠胃癌原位移植模型，采集裸鼠胃癌组织及血液的体内实验与胃癌细胞培养的体外实验，运用生物学技术RT-PCR、MTT、ELISA、Western Blot及RNA干扰等技术探讨Fas-FasL-FADD-Caspase-8通路介导下游细胞凋亡，与TFF3-NF-κB- ERK促进细胞增殖之间的失衡对胃癌形成过程中的病理生理机制，并探讨临床疗效确切的七方胃痛颗粒对胃癌失巢凋亡与增殖的调节效应。.科学意义：本课题从基因转录和蛋白结合位点水平分析了七方胃痛颗粒对胃癌细胞Fas-FasL-FADD-Caspase-8促凋亡通路与TFF3-NF-κB- ERK促细胞增殖通路以及对胃癌SGC7901细胞NF-κB基因沉默模型的影响，国内外很少见利用此技术研究该复方中药作用机制的报道。