PM_(2.5)对人肺癌细胞A549迁移、侵袭能力的增强作用  

Enhancement Effect of PM_(2.5) on Migration and Invasion Abilities of Human Lung Carcinoma Cell Line(A549)

作　　者：杨丹[1] 周伟强[2] 杨彪[2] 肖纯凌[2] 

机构地区：[1]沈阳医学院药理学教研室,沈阳110034 [2]沈阳医学院辽宁省环境污染与微生态重点实验室,沈阳110034

出　　处：《生态毒理学报》2017年第5期243-250,共8页Asian Journal of Ecotoxicology

基　　金：沈阳市科技局计划项目(No.F14-181-1-00);辽宁省自然科学基金计划重点项目(No.20170520037)

摘　　要：为探讨PM_(2.5)(particulate matter 2.5)对人肺癌细胞A549迁移、侵袭能力的影响并探讨相关机制,采用含有不同浓度PM_(2.5)的无血清及抗生素培养液对A549细胞培养72 h,利用MTT法检测A549细胞的增殖抑制率。根据MTT实验结果,选择适当的PM_(2.5)暴露浓度用于后续实验,以细胞划痕实验检测细胞迁移能力,transwell小室实验检测细胞迁移、侵袭能力,Western Blotting法检测细胞周期蛋白D1(cyclin D1)、转录因子snail和slug、基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)、基质金属蛋白酶-9(matrix metalloproteinase-9,MMP-9)及胞核内β-链蛋白(β-catenin)的蛋白表达水平。结果显示:细胞划痕实验结果显示10μg·m L-1PM_(2.5)组A549细胞划痕创面愈合率为(46.34%±5.19%),与对照组比较显著增高(P<0.05);transwell小室法迁移和侵袭实验结果显示10μg·m L-1PM_(2.5)组穿膜细胞数平均为(165.67±6.62)个和(47.83±2.04)个,与对照组比较显著增多(P<0.05);Western Blotting实验结果显示PM_(2.5)(10μg·m L-1)可显著上调A549细胞cyclin D1、snail、slug、MMP-2、MMP-9和胞核内β-catenin蛋白表达水平。因此,PM_(2.5)可通过提高Wnt/β-catenin通路活性及其下游cyclin D1、snail、slug、MMP-2和MMP-9的蛋白表达而增强A549细胞的迁移、侵袭能力。To investigate the effects and mechanisms of PM2.5 on migration and invasion abilities in human lung cell line （A549）, the A549 cells were cultured for 72 hours in the medium containing different concentrations of PM2.5 without serum and antibiotics. The proliferation inhibition rate of A549 cells was detected by MTT assay. According to the experimental results of MTT, the appropriate concentration of PM2.5 exposure was selected for subsequent experiments. The wound-healing assay was performed to observe the cell migration ability and the transwell assay was performed to observe the cell migration and invasion abilities. The protein expression levels of β-catenin in the nucleus, cyclin D1, snail, slug, MMP-2 and MMP-9 were detected by Western Blotting method. The wound-healing assay showed that the wound-healing rate of group treated by PM2.5 at the concentration of 10 μg·mL^-1was （46.34% ± 5.19%）, which was increased signifcantly compared with the control group （P〈0.05）. The transwell assay showed that the transwell cell numbers of group treated by PM2.5 at the concentration of 10 μg·mL^-1were 165.67 ± 6.62 and 47.83 ± 2.04 for migration and invasion experiments respectively, which were increased significantly compared with the control group （P〈0.05）. The Western Blotting assay showed that PM2.5 （10 μg·mL^-1 can significantly up-regulating the expression levels of β-catenin in the nucleus, cyclin D1, snail, slug, MMP-2 and MMP-9 in A549 cells. Thus, PM2.5 can enhance the migration and invasion abilities of A549 cells through up-regulated the activity of Wnt/β-catenin pathway and the protein expression levels of its downstream proteins such as cyclin D1, snail, slug, MMP-2 and MMP-9.

关 键 词：PM2.5 人肺癌细胞 A549 β-catenin snail SLUG 金属蛋白酶-2 金属蛋白酶-9 细胞周期蛋白D1 

分 类 号：X171.5[环境科学与工程—环境科学]