摘要
目的探讨IL-17对胃癌细胞上皮间质转化（EMT）和侵袭迁移的影响及其可能作用机制。方法（1）取对数生长期的胃癌细胞株MGC-803，分别运用浓度为0、1ng/mL、10ng／mL、100ng／mL、1μg/mL IL-17干预48h，观察细胞形态学变化。取细胞形态变化最为明显的浓度作为后续实验最适浓度。浓度为100ngJmL的IL-17干预胃癌细胞48h，设为实验组；加入等量PBS干预胃癌细胞48h，设为对照组。（2）RT—PCR检测两组胃癌细胞中钙粘附蛋白E（E—cadherin）、波形蛋白（Vimentin）的RNA表达水平。（3）Westernblot检测两组胃癌细胞中E-cadherin和Vimentin的相对蛋白表达量。（4）划痕实验和Transwell实验检测两组胃癌细胞的侵袭和迁移能力。正态分布的计量资料采用x±s表示，两组比较采用t检验。结果（1）胃癌细胞EMT形态变化：不同浓度IL-17处理MGC-803胃癌细胞48h后，细胞形态发生显著改变。主要是细胞由多角紧密连接状态逐渐向连接松散，梭形形态转变，细胞粘附能力明显下降，且随着IL-17浓度从0增加至100ng／mL时，细胞形态改变逐渐明显；当浓度达到100ng／mL时，细胞形态改变最明显；但当IL-17浓度继续增加至1μg/mL时，细胞形态改变不再显著，部分细胞出现死亡，漂浮现象。（2）RT—PCR检测结果：实验组和对照组胃癌细胞中E—cadherinmRNA相对表达量分别为0．45±0．13和1．06±0．23；VimentinmRNA相对表达量分别为2．594-0．55和1．23±0．gl，上述指标两组比较，差异均有统计学意义（t=3．811，2．923，P〈0．05）。（3）Westernblot检测结果：实验组和对照组胃癌细胞中E—cadherin蛋白相对表达量分别为0．86±0．17和1．56±0．29；Vimentin蛋白相对表达量分别为1．01±0．12和0．56±0．17，上述指标两组比较，差异均有统计学意义（t=3．551，3．601，P〈0．05）。（4）划痕实验结果显示：划痕56h后，实验组
Objective To investigate the effect of interleukin-17 （IL-17 ） in the gastric cancer cell migration and invasion via regulating epithelial-mesenchymal transition （EMT） and its potential function. Methods （1） Human gastric cancer cell MGC-803 lines in the logarithmic growth phase were stimulated by 0.1ng/mL, 10 ng/mL,100 ng/mL and lp, g/mL of IL-17 for 48 hours, and the phenotypic changes were observed. The concentration of IL-17 was selected for follow-on experiments based on the most obvious phenotypic changes. Gastric cancer cell MGC-803 which were stimulated by 100 ng/mL of IL-17 and PBS for 48 hours were allocated into the experimental group and control group, respectively. （2） The expressions of E-cadherin and Vimentin mRNA in gastric cancer ceils were assayed through real-time PCR （RT-PCR）. （3） The relative expressions of E-cadherin and Vimentin proteins in gastric cancer cells were assayed by the Western blot. （4） The scratch test and Transwell detection were also utilized to study the migration and invasion of gastric cancer cell MGC-803 in vitro. Measurement data with normal distribution were presented as x± s and comparison between groups was analyzed using the t test. Results （ 1 ） There were significant phenotypic changes in the gastric cancer cell after the different concentration of IL-17 stimulated gastric cancer cell MGC-803 for 48 hours. Cells were changed from polygonal and tight junction to spindle and loosely junction with a deterioration of cell adhesion. Cell phenotypes were gradually changed as the concentration of IL-17 was changed from 0 to 100 ng/mL. Phenotypic changes were the most obvious when 100 ng/mL of IL-17 was used, but these were non-significant as the concentration of IL-17 increased to 1 μg/mL with the death and floating of some cells. （2） The relative expressions of E-cadheriu mRNA and Vimentin mRNA in RT-PCR were 0.45 ±0.13 and 1.06 ±0.23 in the experimental group and 2.39±0.55 and 1.23 ±0.41 in the control group, respectively, with
出处
《中华消化外科杂志》
CAS
CSCD
北大核心
2015年第11期948-952,共5页
Chinese Journal of Digestive Surgery
基金
国家自然科学基金（81372559）
关键词
胃肿瘤
白细胞介素17
上皮间质转化
迁移
侵袭
Gastric neoplasms
Interleukin- 17
Epithelial-mesenchymal transition
Migration
Invasion
