摘要
目的探讨沉默高迁移率族蛋白1(HMGB1)基因对非小细胞肺癌细胞上皮间质转化(EMT)的影响及其作用机制。方法取传3代非小细胞肺癌A549细胞,随机分为si-HMGB1组、阴性对照组、空白组。si-HMGB1组、阴性对照组分别转染HMGB1 siRNA序列、阴性对照siRNA序列,空白组不予转染。各组继续孵育72 h,检测HMGB1 mRNA表达。收集各组孵育72 h细胞,采用Western blotting法检测EMT相关蛋白(E-cadherin、VDR、N-cadherin、Vimentin)和TGF-β/Smad信号通路相关蛋白(Snail、NF-κB p65、p-NF-κB p65)表达。结果 si-HMGB1组HMGB1 mRNA相对表达量明显低于阴性对照组和空白组(P均<0.05),而阴性对照组与空白组比较P>0.05。si-HMGB1组E-cadherin、VDR蛋白相对表达量明显高于阴性对照组和空白组(P均<0.05),N-cadherin、Vimentin蛋白相对表达量明显低于阴性对照组和空白组(P均<0.05),而阴性对照组与空白组各EMT相关蛋白相对表达量比较P均>0.05。si-HMGB1组Snail、p-NF-κB p65蛋白相对表达量明显低于阴性对照组和空白组(P均<0.05),而阴性对照组与空白组比较P均>0.05。三组间NF-κB p65蛋白相对表达量比较P均>0.05。结论沉默HMGB1基因能够逆转非小细胞肺癌细胞EMT,其机制可能与阻断Snail/NF-κB信号通路有关。
Objective To explore the effect of high mobility group box 1（ HMGB1） on epithelial mesenchymal transition（ EMT） of non-small-cell lung cancer（ NSCLC） and its mechanism. Methods NSCLC A549 cells in the third generation were randomly divided into the si-HMGB1 group,negative control group,and blank group. Cells in the si-HMGB1 group and negative control group were transfected with HMGB1 small interfering RNA（ siRNA） sequence,HMGB1 negative control siRNA transfection,and cells in the blank group were not transfected. After 72 h,the HMGB1 silencing efficiency of each group was verified. The expression of EMT-related proteins（ E-cadherin,V-cadherin,Vimentin） and signal pathway-related proteins（ Snail,NF-κB p65,p-NF-κB p65） in cells collected after 72-hour transfection was detected by Western blotting. Results The expression of HMGB1 mRNA in the si-HMGB1 group was significantly lower than that in the negative control group and blank group（ P〈0. 05）,and there was no significant difference between the negative control group and the blank group（ P〉0. 05）. The protein expression of E-cadherin and VDR of the si-HMGB1 group was significantly higher,and the protein expression of N-cadherin and Vimentin was significantly lower than that of the negative control group and blank group（ all P〈0. 05）,and there was no significant difference between the negative control group and the blank group（ P〈0. 05）. The expression of Snail,p-NF-κB p65 protein of the si-HMGB1 group was lower than that of the negative control group and the blank group（ P〈0. 05）,and there was no significant difference between the negative control group and the blank group（ P〉0. 05）. No significant difference was found in the expression of p-NF-κB p65 among these three groups（ all P〉0. 05）. Conclusion HMGB1 silencing can reverse the EMT process in A549 cells byblocking the Snail/NF-κB signaling pathway.
出处
《山东医药》
CAS
北大核心
2017年第36期5-8,共4页
Shandong Medical Journal
基金
基金项目:卫生部医药卫生科技发展研究(卫技中[2011]24号)
关键词
非小细胞肺癌
高迁移率族蛋白1
上皮间质转化
基因沉默
non-small-cell lung cancer
high mobility group box 1
epithelial mesenchymal transition
gene silencing
