【作者】 那迪； 宋岩； 姜成钢； 徐盈盈； 鲁翀； 王振宁； 徐惠绵；

【Author】 Di Na;Yan Song;Cheng-Gang Jiang;Ying-Ying Xu;Chong Lu;Zhen-Ning Wang;Hui-Mian Xu;Department of Surgical Oncology, The First Hospital of China Medical University;The Shengjing Hospital of China Medical University;

【机构】 中国医科大学附属第一医院,胃肠肿瘤外科； 中国医科大学附属盛京医院,耳鼻咽喉头颈外科；

【摘要】 腹膜转移是进展期胃癌根治术后最常见的复发形式,是胃癌患者的主要死亡原因。胃癌细胞诱导腹膜间皮细胞的损伤改变很可能与细胞凋亡相关。本研究目的:建立胃癌腹膜转移早期间皮细胞的损伤模型,确定胃癌腹膜转移早期癌细胞对间皮细胞的损伤与细胞凋亡的联系,并探讨凋亡机制。方法:收集处理人胃癌细胞分泌的上清,与人腹膜间皮细胞系HMrSV5共培养。MTT实验观察间皮细胞形态、增殖改变,DNA电泳,透射电镜、扫描电镜确定凋亡形成,流式细胞仪和A0/EB荧光染色判断凋亡比例。C57BL/6小鼠建立腹膜转移模型。结果:通过体内、体外实验发现:接触胃癌上清后,间皮细胞受损脱落,出现大片裸区。胃癌细胞的分泌上清液明显的抑制了间皮细胞的生长。胃癌细胞上清处理后腹膜间皮细胞膨胀变圆,染色质高度凝聚,核膜皱缩,内质网膨胀。部分细胞核解体,核仁消失,胞膜下陷,形成由质膜包裹内含完整的细胞器、核碎片等细胞内容物的凋亡小体,DNA Ladder证实了细胞凋亡的存在。胃癌细胞上清诱导了间皮细胞凋亡抑制细胞增殖,并且程度与时间呈正相关。结论:胃癌细胞在腹膜转移早期可以通过其分泌物诱导间皮细胞损伤、凋亡。提示保护间皮细胞,抗间皮细胞凋亡在胃癌腹膜转移治疗中可能有一定应用价值。更多还原

【Abstract】 Objective:This study was carried out to evaluate the effects of gastric cancer cell supernatant on human peritoneal mesothelial cells viability and apoptosis and to investigate the mechanism of action of gastric cancer in a mesothelial cell line(HMrSV5).Methods:Cell viability was measured using the MTT assay.Mesothelial cells treated with gastric cancer cell supernatant were stained with acridine orange/ethidium bromide and subjected to fluorescence microscopy.C57BL/6 mice were used to establish a cancer invasion model in mice.Results:Morphological changes and exfoliation occurred and naked areas appeared in both cultured mesothelial cells and parietal peritoneum after they were treated with gastric cancer cell supernatant.Cell supernatant from gastric cancer cell induced apoptosis of mesothelial cells in a time-dependent manner.Obvious morphological changes were observed in cell apoptosis,such as condensation of chromatin,nuclear fragmentations and apoptotic ladders.Conclusions:These findings demonstrate that gastric cancer cells can induce apoptosis of human peritoneal mesotfaelial cells through supematants in the early peritoneal metastasis.更多还原