摘要
目的探讨SHP2在幽门螺杆菌脂多糖(Hp-LPS)介导STAT3通路调控IL-6表达的作用。方法 Hp-LPS 0、1、10μg/ml刺激胃癌细胞MKN45 12h,采用RT-PCR检测STAT3、SHP2mRNA表达,Western blot检测STAT3、SHP2蛋白表达。Hp-LPS 1μg/ml刺激胃癌细胞MKN45 0、1、10、30、60min以及Hp-LPS 0、1、10、20、40μg/ml刺激胃癌细胞MKN45 30min时,采用Western blot检测p-STAT3和p-SHP2蛋白表达。ELISA法检测SHP2阻断前后IL-6水平变化。结果STAT 3mRNA相对表达量随Hp-LPS浓度升高而升高(P<0.01),SHP2mRNA相对表达量随Hp-LPS浓度升高而降低(P<0.05)。不同浓度Hp-LPS刺激细胞MKN45的STAT3和SHP2蛋白相对表达量比较差异无统计学意义(P>0.05)。Hp-LPS刺激细胞各时间点p-STAT3蛋白相对表达量差异有统计学意义(P<0.05)。p-STAT 3蛋白相对表达量随着Hp-LPS浓度的增加而增高(P<0.05)。阻断SHP2后的IL-6水平高于阻断前[(15.17±1.16)pg/ml vs.(10.63±0.66)pg/ml](P<0.05)。结论 SHP2是Hp-LPS激活STAT3通路和促进IL-6表达的重要调控因子。
Objective To investigate the role of SHP2 in the regulation of IL-6expression via Hpylori-lipopolysaccharide（Hp-LPS）-mediated STAT3 pathway.Methods The gastric cancer cells MKN45 were stimulated with Hp-LPS in the concentrations of 0,1,and 10μg/ml for 12 hours,after which the mRNA and protein expressions of STAT3 and SHP2 were detected by RT-PCR and Western blot,respectively.The protein expressions of p-STAT3 and p-SHP2 were detected by Western blot at 0,1,10,30,and 60 minutes after the MKN45 cells were stimulated by Hp-LPS 1μg/ml.The protein expressions of p-STAT3 and p-SHP2 were detected by Western blot at 30 minutes after the MKN45 cells were stimulated by Hp-LPS in the concentrations of 0,1,10,20,and 40μg/ml.The IL-6levels were detected by ELISA before and after inhibition of SHP2.Results The STAT3 mRNA expression was increased,while SHP2 mRNA expression was decreased,when the Hp-LPS levels increased（P〈0.01 or P〈0.05）.There were no significant differences in STAT3 and SHP2 protein expressions after the MKN45 cells were stimulated by different concentrations of Hp-LPS（P〉0.05）.The p-STAT3 protein expressions were significant different after the MKN45 cells were stimulated by Hp-LPS for different times（P〈0.05）.The p-STAT3 protein expression was increased as the Hp-LPS concentration increased（P〈0.05）.The IL-6level was higher after the inhibition of SHP2 than that before[（15.17±1.16）pg/ml vs.（10.63±0.66）pg/ml]（P〈0.05）.Conclusion SHP2 is an important regulator in the process of Hp-LPS activating STAT3 to induce IL-6expression.
作者
唐春丽
叶峰
张国新
TANG Chunli YE Feng ZHANG Guoxin(Department of Gastroenterology, People ＇ s Hospital of Suqian , Suqian 223800, CHIN)
出处
《江苏医药》
CAS
2017年第6期381-383,共3页
Jiangsu Medical Journal
基金
国家自然科学基金(81072032)
