{"title":"脱氧胆酸盐诱导培养的正常人食管粘膜上皮细胞凋亡。","authors":"Ru Zhang, Jun Gong, Hui Wang, Li Wang, Li-wei Ran","doi":"","DOIUrl":null,"url":null,"abstract":"<p><strong>Objective: </strong>To study the effect of deoxycholate in inducing apoptosis of human normal esophageal mucosal epithelial cells in vitro and investigate the molecular mechanism.</p><p><strong>Methods: </strong>Cultured normal human esophageal mucosal epithelial cells were treated with deoxycholate, and the cell apoptosis were evaluated with TUNEL, DNA ladder, flow cytometry with PI-staining, Annexin V-FITC conjugated with PI staining, and Western blotting.</p><p><strong>Results: </strong>Flow cytometry, TUNEL and DNA ladder demonstrated that deoxycholate could induce apoptosis in normal human esophageal mucosal epithelial cells in a dose- and time-dependent manner. A percentage of 21.3% of the cell population treated with deoxycholate at 500 micromol/L for 30 min exhibited detectable caspase-3 activity shown by flow cytometry, which was significantly higher than the control level (1.5%, P<0.01). Western blotting suggested that deoxycholate down-regulated Bcl-2 protein expression and up-regulated Bax expression, but Fas was negative in the cells before and after deoxycholate treatment.</p><p><strong>Conclusions: </strong>Deoxycholate could induce apoptosis in cultured human esophageal mucosal epithelial cells. Aaspase-3 activation, Bcl-2 protein down-regulation and Bax up-regulation are involved in deoxycholate-induced apoptosis, which does not involve Fas-L/Fas.</p>","PeriodicalId":11097,"journal":{"name":"Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2005-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Deoxycholate induces apoptosis in cultured normal human esophageal mucosal epithelial cells].\",\"authors\":\"Ru Zhang, Jun Gong, Hui Wang, Li Wang, Li-wei Ran\",\"doi\":\"\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><strong>Objective: </strong>To study the effect of deoxycholate in inducing apoptosis of human normal esophageal mucosal epithelial cells in vitro and investigate the molecular mechanism.</p><p><strong>Methods: </strong>Cultured normal human esophageal mucosal epithelial cells were treated with deoxycholate, and the cell apoptosis were evaluated with TUNEL, DNA ladder, flow cytometry with PI-staining, Annexin V-FITC conjugated with PI staining, and Western blotting.</p><p><strong>Results: </strong>Flow cytometry, TUNEL and DNA ladder demonstrated that deoxycholate could induce apoptosis in normal human esophageal mucosal epithelial cells in a dose- and time-dependent manner. A percentage of 21.3% of the cell population treated with deoxycholate at 500 micromol/L for 30 min exhibited detectable caspase-3 activity shown by flow cytometry, which was significantly higher than the control level (1.5%, P<0.01). Western blotting suggested that deoxycholate down-regulated Bcl-2 protein expression and up-regulated Bax expression, but Fas was negative in the cells before and after deoxycholate treatment.</p><p><strong>Conclusions: </strong>Deoxycholate could induce apoptosis in cultured human esophageal mucosal epithelial cells. Aaspase-3 activation, Bcl-2 protein down-regulation and Bax up-regulation are involved in deoxycholate-induced apoptosis, which does not involve Fas-L/Fas.</p>\",\"PeriodicalId\":11097,\"journal\":{\"name\":\"Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2005-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Di 1 jun yi da xue xue bao = Academic journal of the first medical college of PLA","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0
摘要
目的:研究脱氧胆酸盐在体外诱导人正常食管粘膜上皮细胞凋亡中的作用,并探讨其分子机制。方法:用脱氧胆酸盐处理培养的正常人食管粘膜上皮细胞,采用TUNEL、DNA阶梯、PI-染色流式细胞术、Annexin V-FITC - PI染色和Western blotting检测细胞凋亡情况。结果:流式细胞术、TUNEL和DNA阶梯实验显示脱氧胆酸盐能诱导正常人食管粘膜上皮细胞凋亡,且呈剂量和时间依赖性。500微mol/L脱氧胆酸处理30 min后,流式细胞术检测到21.3%的细胞群显示caspase-3活性,显著高于对照(1.5%)。结论:脱氧胆酸盐可诱导培养的人食管粘膜上皮细胞凋亡。脱氧胆碱诱导的细胞凋亡与Aaspase-3激活、Bcl-2蛋白下调和Bax蛋白上调有关,与Fas- l /Fas无关。
[Deoxycholate induces apoptosis in cultured normal human esophageal mucosal epithelial cells].
Objective: To study the effect of deoxycholate in inducing apoptosis of human normal esophageal mucosal epithelial cells in vitro and investigate the molecular mechanism.
Methods: Cultured normal human esophageal mucosal epithelial cells were treated with deoxycholate, and the cell apoptosis were evaluated with TUNEL, DNA ladder, flow cytometry with PI-staining, Annexin V-FITC conjugated with PI staining, and Western blotting.
Results: Flow cytometry, TUNEL and DNA ladder demonstrated that deoxycholate could induce apoptosis in normal human esophageal mucosal epithelial cells in a dose- and time-dependent manner. A percentage of 21.3% of the cell population treated with deoxycholate at 500 micromol/L for 30 min exhibited detectable caspase-3 activity shown by flow cytometry, which was significantly higher than the control level (1.5%, P<0.01). Western blotting suggested that deoxycholate down-regulated Bcl-2 protein expression and up-regulated Bax expression, but Fas was negative in the cells before and after deoxycholate treatment.
Conclusions: Deoxycholate could induce apoptosis in cultured human esophageal mucosal epithelial cells. Aaspase-3 activation, Bcl-2 protein down-regulation and Bax up-regulation are involved in deoxycholate-induced apoptosis, which does not involve Fas-L/Fas.