杨丹¹,
周伟强²,
杨彪²,
肖纯凌²
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

作者简介: 杨丹(1978-),女,博士,讲师,研究方向为肿瘤学,E-mail:Ld_love415@163.com.

基金项目: 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)


中图分类号: X171.5

Enhancement Effect of PM_2.5 on Migration and Invasion Abilities of Human Lung Carcinoma Cell Line (A549)

Yang Dan¹,
Zhou Weiqiang²,
Yang Biao²,
Xiao Chunling²
1. Department of Pharmacology, Shenyang Medical College, Shenyang 110034, China;
2. Key Laboratory of Environmental Pollution and Microecology of Liaoning Province, Shenyang Medical College, Shenyang 110034, China

CLC number: X171.5

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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10 μg·mL^-1 PM_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10 μg·mL^-1 PM_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(10 μg·mL^-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。
关键词: PM_2.5/
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

Abstract:To investigate the effects and mechanisms of PM_2.5 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 PM_2.5 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 PM_2.5 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 PM_2.5 at the concentration of 10 μg·mL^-1 was (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 PM_2.5 at the concentration of 10 μg·mL^-1 were 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 PM_2.5 (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, PM_2.5 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.
Key words:PM_2.5/
A549/
β-catenin/
snail/
slug/
MMP-2/
MMP-9/
cyclin D1/