异丙酚通过miR-218抑制人食管鳞癌细胞系KYSE150的侵袭和迁移

目的探讨异丙酚对人食管鳞癌细胞系KYSE150侵袭和迁移的影响及其机制。方法以0、2.5、5和10μg/L异丙酚处理KYSE150细胞,24 h后分别采用Transwell小室法和划痕实验检测细胞的侵袭和迁移能力;RT-qPCR检测细胞中miR-218的表达;Western blot检测HMGB1蛋白的表达;生物信息学软件预测和双荧光素酶报告基因实验检测miR-218和HMGB1的靶向关系,观察miR-218对HMGB1蛋白的调控作用。采用脂质体法转染miR-218抑制剂或pcDNA3.1-HMGB1-GFP过表达载体质粒后,观察miR-218和HMGB1蛋白的表达以及下调miR-218或上调HMGB1表达对10μg/L异丙酚处理的KYSE150细胞侵袭和迁移的影响。结果异丙酚呈浓度依赖性地抑制KYSE150细胞侵袭、迁移和细胞中HMGB1蛋白的表达,并促进miR-218的表达(P<0.05)。双荧光素酶报告基因实验证实HMGB1是miR-218的靶基因,miR-218可负向调控HMGB1蛋白的表达(P<0.05)。下调miR-218表达可逆转异丙酚对KYSE150细胞侵袭和迁移的抑制作用(P<0.05);同时上调HMGB1表达可逆转异丙酚对KYSE150细胞侵袭和迁移的抑制作用(P<0.05)。结论异丙酚抑制食管鳞癌KYSE150细胞侵袭和迁移,其作用机制可能与miR-218靶向调控HMGB1有关。

Propofol inhibits invasion and migration of esophageal squamous carcinoma cell line KYSE150 through miR-218

Objective To investigate the effect of propofol on invasion and migration of esophageal squamous cancer cell line KYSE150 and potential mechanism.Methods KYSE150 cells were treated with 0, 2.5, 5 and 10 μg/L propofol,Transwell chamber and scratch assay were used to detect the invasion and migration of cells after 24 h. RT-qPCR was used to detect the expression of miR-218 in cells. Western blot analysis was performed to detect the expression of HMGB1 protein. Bioinformatics software prediction and dual luciferase reporter gene assay were performed to detect the targeting relationship between miR-218 and HMGB1, Western blot to detect the regulation of miR-218 on HMGB1 protein. After transfection of miR-218 inhibitor or pcDNA3.1-HMGB1-GFP over-expression vector plasmid was constructed by liposome method, the expression of miR-218 was detected by RT-qPCR, the expression of HMGB1 protein was detected by Western blot, Transwell chamber and scratch test. The effect of down-regulating miR-218 or up-regulating HMGB1 expression on invasion and migration of 10 μg/L propofol-treated KYSE150 cells was examined. Results Propofol inhibited the invasion, migration and expression of HMGB1 proteinin KYSE150 cells in a concentration-dependent manner and promoted the expression of miR-218. Dual luciferase reporter gene experiments confirmed that HMGB1 was a target gene of miR-218, and Western blot analysis confirmed that miR-218 negatively regulated the expression of HMGB1 protein. Down-regulation of miR-218 expression reversed the inhibitory effect of propofol on invasion and migration of KYSE150 cells. Up-regulation of HMGB1 expression reversed the inhibitory effect of propofol on invasion and migration of KYSE150 cells. Conclusions miR-218 plays an active role in the inhibition of invasion and migration of esophageal squamous cell carcinoma KYSE150 cells by propofol, and its mechanism is potentially related to targeted regulation of HMGB1.