摘要
目的研究转化生长因子β1(TGF-β1)对人胃癌细胞BGC-823侵袭性的影响。方法将体外培养的人胃癌细胞BGC-823用外源性人重组TGF-β1干预后,用四甲基偶氮唑蓝比色法(MTT法)检测TGF-β1对细胞增殖的影响;免疫细胞化学法、免疫荧光法、流式细胞术分别检测Snail、E-cadher-in及N-cadherin蛋白的表达;Transwell侵袭实验检测TGF-β1对细胞侵袭性的影响。结果①经过18h的作用,低浓度(1、5和10ng/ml)的TGF-β1促进胃癌细胞的增殖,高浓度(20和50ng/ml)的TGF-β1则抑制胃癌细胞的增殖,且呈剂量依赖性;选取不同时间段(18、42、66h)检测细胞生长曲线,发现不呈时间依赖性;②在外源性TGF-β1作用下,胃癌细胞均不同程度地表达Snail、E-cadherin及N-cadherin,其中TGF-β1对Snail、N-cadherin表达有促进作用,而对E-cadher-in的表达有抑制作用,E-cadherin和N-cadherin的表达呈相反的趋势;③选取不同浓度的TGF-β1(1、10和20ng/ml),发现10ng/ml的TGF-β1明显促进胃癌细胞的侵袭性(P<0.05)。结论不同浓度的TGF-β1对胃癌细胞增殖的影响不同;TGF-β1有增加胃癌细胞侵袭性的作用,该作用可能是通过上皮间质转化而产生,钙黏蛋白表达的变化可能是重要的分子学机制之一。
Objective To investigate the influence of TGF-β1on invasive ability of the gastric adenocarcinoma cell line BGC-823 in vitro. Methods Human low-differentiated gastric adenocarcinoma cell line BGC-823 was cultured in vitro. After exposed to different concentration of TGF-β1, the proliferation of BGC-823 cells was measured by MTT assay. The expression of Snail, E-cadherin and N-eadherin was detected by immunocytochemical staining （ ICC）, immunofluorescence assay （IFA） and flow cytometry （FCM）. The changes of invasion ability of BGC-823 were detected by transwell cell culture chambers. Results I. After exposured to TGF-β1 （ 1, 5 and 10 ng/ml） for 18 h, the growth of BGC-823 cells was promoted dose-dependently. Meanwhile the inhibitory effect of TGF-β1 （20 and 50 ng/ml） was observed unexpectedly. Then the cell growth curve was measured at different time （18, 42, 66 h）. As a result, it didn＇t show time-dependent manner. 2. The expression of Snail, E-eadherin and N-cadherin was measured at different levels after culture in the presence of human recombinant TGF-β1. The opposite trend was showed between E-cadherin and N-eadherin. TGF-β1 inhibited the expression of E-eadherin, but enhanced that of N-cadherin and Snail. 3. The changes of invasion were detected in BGC-823 cells exposed to TGF-β1 （ 1, 10 and 20 ng/ml）, the rate of the changes was significantly high at dose of 10 ng/ml. Conclusion The proliferation of BGC-823 cells is determined by different concentrations of TGF-β1. The TGF-β1 does have the ability to activate EMT in gastric adenocarcinoma cells, thus increase their invasion. The eadherins switch is one of the important molecular mechanisms in this phenomenon.
出处
《安徽医科大学学报》
CAS
北大核心
2009年第1期18-22,共5页
Acta Universitatis Medicinalis Anhui
基金
安徽省教育厅重点科研项目基金(编号:kj2007A029)
关键词
胃肿瘤/病理学
转化生长因子Β
肿瘤细胞
培养
stomach neoplasms/pathology
transforming growth factor beta
tumor cells, cultured
