摘要
目的探讨亮菌多糖(ATPS)对人胃黏膜上皮细胞(GES-1)发生上皮-间质转化(EMT)的影响及相关机制。方法利用IL-8诱导GES-1细胞建立EMT模型,实验设空白对照组、模型组(IL-8诱导组)、实验组[ATPS低剂量组(IL-8诱导1 h后加入ATPS 30 mg/L)、ATPS中剂量组(IL-8诱导1 h后加入ATPS 60 mg/L)、ATPS高剂量组(IL-8诱导1 h后加入ATPS 120 mg/L)]。应用光学显微镜观察GES-1细胞形态学变化,Western blot法检测EMT相关蛋白及PDCD4蛋白表达水平,MTT法检测不同浓度ATPS对GES-1细胞存活率影响,细胞划痕实验检测各组GES-1细胞侵袭能力变化。结果光学显微镜观察到IL-8作用24 h后GES-1细胞形态由不规则多边形向梭形转化,细胞排列紊乱,侵袭能力增强,不同剂量亮菌多糖作用后细胞形态可部分恢复,侵袭能力降低。Western blot结果提示:与模型组相比,实验组上皮标记物E-钙黏蛋白增多(P<0.05),间质标志物N-钙黏蛋白、波形蛋白减少(P<0.05),PDCD4表达增多(P<0.05)。细胞划痕实验观察到模型组GES-1细胞与空白组相比侵袭扩散能力明显增强,实验组与模型组相比侵袭扩散能力明显减弱。结论 ATPS可通过抑制GES-1细胞EMT的发生逆转胃癌前病变,其机制可能与促进抑癌基因PDCD4表达有关。
Objective To investigate the influence of Armillariella polysaccharide（ ATPS） on epithelial-mesenchymal transition（ EMT） in gastric epithelial cells（ GES-1） and its mechanism. Methods Construction of EMT model with GES-1 cells induced by IL-8. The blank control group,model group（ IL-8 group induced group）,experimental group [low dose group（ IL-8 inducted 1 h,ATPS 30 mg/L）,middle dose of ATPS group（ IL-8 inducted 1 h,ATPS 60 mg/L）,high dose group（ IL-8 inducted 1 h,ATPS 120 mg/L） ]. Morphological changes of GES-1 cells were observed by optical microscope,Western blot was used to detect the expression levels of EMT-related protein and PDCD4 protein,MTT assay was used to detect the effect of different concentrations of ATPS on the survival rate of GES-1 cells,and cell scratch test was used to detect the invasion ability of GES-1 cells in each group. Results After treatment with IL-8 for 24 hours,the morphological changes of GES-1 cells were observed from irregular polygons to spindle,cell arrangement disorder,invasive ability increased,the morphology of the cells could be partially restored and invasive ability weakened after the action of different doses of ATPS. Western blot results showed that the experimental group the E-cadherin and PDCD4 were increased（ P 〈 0. 05） as well as the N-cadherin and Vimentin were reduced（ P 〈 0. 05） compared with the model group. Cell scratch assay showed that the model group of GES-1 cells compared with the blank group significantly increased the ability of invasion and proliferation,experimental group compared with the model group significantly reduced the ability of invasion and proliferation. Conclusion ATPS can reverse the precancerous lesion by inhibiting the occurrence of EMT in GES-1 cells,and its mechanism may be related to ATPS promoting the expression of PDCD4.
作者
赵莉莉
朱耀东
李平
张梅
Zhao Lili;Zhu Yaodong;Li Ping(Dept of Integrated Chinese and Western Oncology,The First Affiliated Hospital of Anhui Medical University,Hefei 23002)
出处
《安徽医科大学学报》
CAS
北大核心
2018年第6期868-874,共7页
Acta Universitatis Medicinalis Anhui
基金
国家自然科学基金(编号:81603439)
安徽省中医药科技项目(编号:2016zy86)
