基于TGF-β1/Smads通路探讨半夏泻心汤干预胃癌外泌体诱发腹膜间皮细胞EMT阻抑胃癌腹膜转移的作用机制

EMT of peritoneal skin cells is beneficial to gastric cancer peritoneum metastasis, and TGF- β1/Smads is the key pathway to regulate the EMT of cells. Recent studies have found that exosomes of gastric cancer are important ways to induce the EMT of peritoneal skin cells, and explore its mechanism and intervention methods as a new strategy to prevent and control the peritoneal metastasis of gastric cancer. Based on the previous work, this paper puts forward the working hypothesis of "Banxiaxiexin Decoction regulate and control TGF- β1/Smads signaling pathway interferes with the external secretion of the peritoneal epithelial cell EMT and inhibits the peritoneal metastasis of gastric cancer." Research will peritoneal mesothelium cell and cancer of the stomach secrete body non-contact cocultivation, observation of peritoneal mesothelium cell with cancer of the stomach secrete intake and absorb the stomach secrete body after peritoneal mesothelium cell TGF - β1 / Smads access key signaling molecule, EMT interstitial phenotypic marker expression level and the change of cell morphology, the ability of invasion and metastasis of gastric cancer cells, and explore the intervention effect of Banxiaxiexin Decoction and mechanism of action, interpretation of the stomach secrete outside body induced peritoneal mesothelium cell EMT, promote gastric cancer peritoneal metastasis ".Spleen deficient phlegm junction, cold and heat mixed" theory of TCM scientific connotation. This study will provide new ideas and methods for the exploration of the EMT of the peritoneal skin cells induced by the external secretion of traditional Chinese medicine in gastric cancer and the inhibition of the peritoneal metastasis of gastric cancer.

腹膜间皮细胞EMT可促进胃癌腹膜转移。最新研究发现胃癌外泌体是诱发腹膜间皮细胞EMT的重要途径，TGF-β1/Smads信号通路与细胞EMT密切相关，探寻其作用机制和干预手段成为防治胃癌腹膜转移的关键策略之一。结合前期工作基础，提出了“半夏泻心汤调控TGF-β1/Smads信号通路干预胃癌外泌体诱发腹膜间皮细胞EMT，阻抑胃癌腹膜转移”的工作假说。研究拟将腹膜间皮细胞与胃癌外泌体进行非接触共培养，观察腹膜间皮细胞对胃癌外泌体的摄取及摄取胃癌外泌体后腹膜间皮细胞TGF-β1/Smads通路关键信号分子、EMT间质表型标志物表达水平及其细胞形态、胃癌细胞侵袭转移能力的变化，探讨半夏泻心汤的干预作用及作用机制，阐释胃癌外泌体诱发腹膜间皮细胞EMT，促进胃癌腹膜转移“脾虚痰结，寒热错杂”理论的科学内涵。本研究将为探索中医药干预胃癌外泌体诱发腹膜间皮细胞EMT，阻抑胃癌腹膜转移提供新的思路与方法。

在胃癌细胞发生腹膜转移前，腹膜间皮细胞已经发生了EMT。而胃癌外泌体能够诱导腹膜间皮细胞发生EMT。TGF-β1/Smads信号通路与细胞EMT密切相关。临床研究表明半夏泻心汤在治疗胃癌、食管癌等消化道肿瘤方面取得了较好的疗效。我们前期研究也表明，半夏泻心汤含药血清可保护胃癌上清液对腹膜间皮细胞的损伤，抑制人胃癌腹膜高转移潜能细胞的增殖及侵袭转移。因此，我们推测，半夏泻心汤可能调控TGF-β1/Smads信号通路干预胃癌外泌体诱发腹膜间皮细胞EMT，阻抑胃癌腹膜转移。为了验证该假说，我们采用Transwell培养小室将人腹膜间皮细胞与选择的胃癌外泌体进行非接触共培养，观察腹膜间皮细胞对胃癌外泌体的摄取及摄取胃癌外泌体后腹膜间皮细胞TGF-β1/Smads信号通路关键信号分子TGF-β1、Smad2、Smad3、p-Smad2、p-Smad3及Smad7的表达及其细胞形态、EMT间质表型分子标志物蛋白表达、胃癌细胞的粘附侵袭能力的变化，同时观察半夏泻心汤的干预作用。研究表明：胃癌外泌体能够被腹膜间皮细胞摄取，并通过激活腹膜间皮细胞中TGF-β1/Smads信号通路诱发其发生EMT，产生损伤，使其基质破坏，LAMC1和FN1蛋白表达升高，形成腹膜转移前微环境，促进胃癌向腹膜的侵袭，加重胃癌腹膜转移进程。半夏泻心汤作为“辛开苦降”法代表方剂，针对胃癌外泌体诱发腹膜间皮细胞损伤，发生EMT的这一过程，可有效阻抑胃癌外泌体进入腹膜间皮细胞，防止腹膜间皮细胞进一步发生EMT，降低胃癌细胞向腹膜间皮细胞的黏附侵袭，阻抑这一生物学进展。