[Effect of Brucea javanica oil emulsion on proliferation, migration and autophagy of non-small cell lung cancer A549 cells and the underlying mechanisms]

Zhong Nan Da Xue Xue Bao Yi Xue Ban. 2018 Nov 28;43(11):1202-1208. doi: 10.11817/j.issn.1672-7347.2018.11.006.
[Article in Chinese]

Abstract

To investigate the effect of Brucea javanica oil emulsion on proliferation, migration and autophagy of non-small cell lung cancer A549 cells. Methods: First, A549 cells were divided into a control group and a low, medium or high dose of Brucea javanica oil emulsion groups (0, 2.5, 5.0 or 10.0 mg/mL); then, the cells were divided into a 3-MA+Brucea javanica oil emulsion group and a Brucea javanica oil emulsion group in the presence or absence of 3-methyl adenine (3-MA). Cell counting kit-8 (CCK-8) and clone formation assay were used to detect cell proliferation, while the wound scratch and Transwell assay were used to measure cell migration. Cell immunofluorescence and Western blot were used to analyze autophagy. Results: Compared with the control group, the numbers of cell proliferation and colony-formation, the relative cell migration rate and numbers of trans-membrane cells were reduced in a dose-dependent manner in the Brucea javanica oil emulsion groups (all P<0.05). Meanwhile, compared with the control group, the aggregation of microtubule associated protein 1 light chain3 (LC3) green fluorescence and the LC3-II/LC3-I ratios were increased, while p62 level was decreased (all P<0.05) in the high dose group. Compared with the Brucea javanica oil emulsion group (5.0 mg/mL), the cell proliferation, numbers of cell clone formation, cell migration rate and numbers of Transwell transmembrane cells were increased in the 3-MA+Brucea javanica oil emulsion group (all P<0.05). Conclusion: Brucea javanica oil emulsion can promote the autophagy of non-small cell lung cancer A549 cells and inhibit the cell proliferation and migration.

目的:探讨鸦胆子油乳对人非小细胞肺癌A549细胞增殖、迁移和自噬的影响及机制。方法:以A549细胞为研究对象,先采用不同浓度的鸦胆子油乳处理,并将其分为对照组和低、中、高剂量鸦胆子油乳组(0,2.5,5.0,10.0 mg/mL);再采用5.0 mg/mL鸦胆子油乳或3-甲基腺嘌呤(3-methyladenine,3-MA)+5.0 mg/mL鸦胆子油乳处理,并将其分为鸦胆子油乳组和3-MA+鸦胆子油乳组。采用细胞计数试剂盒(cell counting kit-8,CCK-8)和克隆形成实验检测细胞增殖,划痕实验和Transwell实验测量细胞迁移;应用细胞免疫荧光和Western印迹分析细胞自噬。结果:与对照组(0 mg/mL)相比,低、中、高剂量(2.5,5.0,10.0 mg/mL)鸦胆子油乳组的A549细胞增殖受抑制、细胞克隆形成数减少、细胞迁移率降低、Transwell穿膜细胞数减少(均P<0.05),且呈现剂量依赖性(均P<0.05);与对照组(0 mg/mL)相比,5.0 mg/mL鸦胆子油乳组的A549细胞微管相关蛋白轻链3(microtu-bule associated protein 1 light chain 3,LC3)绿色荧光聚集、LC3-II/LC3-I比值增加,p62水平降低(均P<0.05);与鸦胆子油乳组(5.0 mg/mL)相比,3-MA+鸦胆子油乳组的A549细胞增殖增加,细胞克隆形成数、细胞迁移率和Transwell穿膜细胞数增多(均P<0.05)。结论:鸦胆子油乳可诱导非小细胞肺癌A549细胞自噬,可参与肿瘤细胞增殖和迁移的抑制作用。.

MeSH terms

  • A549 Cells
  • Autophagy / drug effects*
  • Brucea* / chemistry
  • Carcinoma, Non-Small-Cell Lung*
  • Cell Movement / drug effects
  • Cell Proliferation / drug effects
  • Humans
  • Lung Neoplasms*
  • Plant Oils* / pharmacology

Substances

  • Plant Oils