Abstract
Exposure to airborne fine particulate matter (PM2.5) induced various adverse health effects, such as metabolic syndrome, systemic inflammation, and respiratory disease. Many works have studied the effects of PM2.5 exposure on cells through intracellular proteomics analyses. However, changes of the extracellular proteome under PM2.5 exposure and its correlation with PM2.5-induced cytotoxicity still remain unclear. Herein, the cytotoxicity of PM2.5 on normal human bronchial epithelia cells (BEAS-2B cells) was evaluated, and the secretome profile of BEAS-2B cells before and after PM2.5 exposure was investigated. A total of 83 proteins (58 upregulated and 25 downregulated) were differentially expressed in extracellular space after PM2.5 treatment. Notably, we found that PM2.5 promoted the release of several pro-apoptotic factors and induced dysregulated secretion of extracellular matrix (ECM) constituents, showing that the abnormal extracellular environment attributed to PM2.5-induced cell damage. This study provided a secretome data for the deep understanding of the molecular mechanism underlying PM2.5-caused human bronchial epithelia cell damage.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Data availability
All data generated or analyzed during this study are included in this published article.
Abbreviations
- AM:
-
acetyl methoxy methyl ester
- ANOVA:
-
one-way analysis of variance
- BCA:
-
bicinchoninic acid
- BEAS-2B cells:
-
bronchial epithelia cells
- BEBM:
-
stable isotope labeling with amino acids in cell culture
- CCK-8:
-
cell counting kit-8
- DCFH/DA:
-
2′,7′-dichlorodihydrofluorescein diacetate
- DKK1:
-
dickkopf1
- DSG2:
-
Desmoglein-2
- ECM:
-
extracellular matrix
- ECM1:
-
extracellular matrix protein-1
- EVs:
-
extracellular vesicles
- FASP:
-
filter-aided sample preparation
- FBS:
-
fetal bovine serum
- FDR:
-
false discovery rate
- TITC:
-
fluorescein isothiocyanate isomer
- GAS1:
-
growth arrest-specific protein 1
- GO:
-
Gene Ontology
- HCD:
-
high-energy collision dissociation
- HRP:
-
horseradish peroxidase
- IC50:
-
half-maximal inhibitory concentration
- ICP-MS :
-
inductively coupled plasma mass spectrometry
- KEGG:
-
Kyoto encyclopedia of genes and genomes
- LAMA4:
-
laminin all subunit alpha-4
- MAPK pathway:
-
mitogen-activated protein kinase pathway
- MIF:
-
migration inhibitory factor
- MLEFF:
-
a strategy combing metabolic labeling, protein “equalization,” protein fractionation, and filter-aided sample preparation
- MMPs:
-
matrix metalloproteinases
- nano-RPLC-MS/MS:
-
nano reverse phase liquid chromatography mass spectrometry
- PAHs:
-
polycyclic aromatic hydrocarbons
- PAI1:
-
plasminogen activator inhibitor type-1
- PBS:
-
phosphate buffered solution
- PCDD/Fs:
-
polychlorinated dibenzo-p-dioxins and dibenzofurans
- PD:
-
proteome discoverer
- PI:
-
propidium iodide
- PI3K/AKT pathway:
-
phosphatidylinositol-3-kinase/AKT serine/threonine kinase pathway
- PVDF:
-
polyvinylidene fluoride
- RNA:
-
ribonucleic acid
- ROS:
-
reactive oxygen species
- SDS:
-
sodium dodecyl sulfate
- TFs:
-
transcription factors
- TNF-alpha:
-
tumor necrosis factor alpha
- VTN:
-
vitronectin
References
Adam M, Schikowski T, Carsin AE, Cai Y, Jacquemin B et al (2015) Adult lung function and long-term air pollution exposure. ESCAPE: a multicentre cohort study and meta-analysis. Eur Respir J 45:38–50. https://doi.org/10.1183/09031936.00130014
Bae S, Pan X-C, Kim S-Y, Park K, Kim Y-H et al (2010) Exposures to particulate matter and polycyclic aromatic hydrocarbons and oxidative stress in schoolchildren. Environ Health Perspect 118:579–583. https://doi.org/10.1289/ehp.0901077
Bo Y, Chang L-Y, Guo C, Lin C, Lau AKH et al (2021) Reduced ambient PM2.5, better lung function, and decreased risk of chronic obstructive pulmonary disease. Environ Int 156:106706. https://doi.org/10.1016/j.envint.2021.106706
Chen H, Liu X, Gao X, Lv Y, Zhou L et al (2021) Epidemiological evidence relating risk factors to chronic obstructive pulmonary disease in China: a systematic review and meta-analysis. PLoS One 16:e0261692. https://doi.org/10.1371/journal.pone.0261692
Cheng L, Hill AF (2022) Therapeutically harnessing extracellular vesicles. Nat Rev Drug Discov. https://doi.org/10.1038/s41573-022-00410-w
Circu ML, Aw TY (2010) Reactive oxygen species, cellular redox systems, and apoptosis. Free Radic Biol Med 48:749–762. https://doi.org/10.1016/j.freeradbiomed.2009.12.022
Ćwieląg-Drabek M, Parascandola M, Piekut A, Spychała A, Dziubanek G et al (2020) Non-dioxin-like PCBs – the key air pollutant associated with lung cancer in 15 cities in Silesia. Pol J Environ Stud 29:1111–1117. https://doi.org/10.15244/pjoes/102786
Dagher Z, Garcon G, Billet S, Gosset P, Ledoux F et al (2006) Activation of different pathways of apoptosis by air pollution particulate matter (PM2.5) in human epithelial lung cells (L132) in culture. Toxicology 225:12–24. https://doi.org/10.1016/j.tox.2006.04.038
Deng X, Zhang F, Rui W, Long F, Wang L et al (2013) PM2.5-induced oxidative stress triggers autophagy in human lung epithelial A549 cells. Toxicol in Vitro 27:1762–1770. https://doi.org/10.1016/j.tiv.2013.05.004
Duan R, Niu H, Yu T, Huang K, Cui H et al (2021) Adverse effects of short-term personal exposure to fine particulate matter on the lung function of patients with chronic obstructive pulmonary disease and asthma: a longitudinal panel study in Beijing, China. Environ Sci Pollut Res 28:47463–47473. https://doi.org/10.1007/s11356-021-13811-y
Forder A, Hsing C-Y, Trejo Vazquez J, Garnis C (2021) Emerging role of extracellular vesicles and cellular communication in metastasis. Cells 10:3429. https://doi.org/10.3390/cells10123429
Fouchecourt MO, Berny P, Riviere JL (1998) Bioavailability of PCBs to male laboratory rats maintained on litters of contaminated soils: PCB burden and induction of alkoxyresorufin O-dealkylase activities in liver and lung. Arch Environ Contam Toxicol 35:680–687. https://doi.org/10.1007/s002449900431
Freeberg MAT, Farhat YM, Easa A, Kallenbach JG, Malcolm DW et al (2018) Serpine1 knockdown enhances MMP activity after flexor tendon injury in mice: implications for adhesions therapy. Sci Rep 8:1–13. https://doi.org/10.1038/s41598-018-24144-1
Heinbockel L, Marwitz S, Schromm AB, Watz H, Kugler C et al (2018) Identification of novel target genes in human lung tissue involved in chronic obstructive pulmonary disease. Int J Chron Obstruct Pulmon Dis 13:2255–2259. https://doi.org/10.2147/copd.s161958
Holowka T, Bucala R (2020) Role of host and parasite MIF cytokines during Leishmania infection. Trop Med Infect Disease 5:46. https://doi.org/10.3390/tropicalmed5010046
Hong Z, Guo Z, Zhang R, Xu J, Dong W et al (2016) Airborne fine particulate matter induces oxidative stress and inflammation in human nasal epithelial cells. Tohoku J Exp Med 239:117–125. https://doi.org/10.1620/tjem.239.117
Hu R, Xie XY, Xu SK, Wang YN, Jiang M et al (2017) PM2.5 Exposure elicits oxidative stress responses and mitochondrial apoptosis pathway activation in HaCaT keratinocytes. Chin Med J 130:2205–2214. https://doi.org/10.4103/0366-6999.212942
Hynes RO (2002) Integrins: bidirectional, allosteric signaling machines. Cell 110:673–687. https://doi.org/10.1016/S0092-8674(02)00971-6
Johnson TM, Yu ZX, Ferrans VJ, Lowenstein RA, Finkel T (1996) Reactive oxygen species are downstream mediators of p53-dependent apoptosis. Proc Natl Acad Sci 93:11848. https://doi.org/10.1073/pnas.93.21.11848
Kang X, Li N, Wang M, Boontheung P, Sioutas C et al (2010) Adjuvant effects of ambient particulate matter monitored by proteomics of bronchoalveolar lavage fluid. Proteomics 10:520–531. https://doi.org/10.1002/pmic.200900573
Kiedrzyńska E, Jóźwik A, Kiedrzyński M, Zalewski M (2014) Hierarchy of factors exerting an impact on nutrient load of the Baltic Sea and sustainable management of its drainage basin. Mar Pollut Bull 88:162–173. https://doi.org/10.1016/j.marpolbul.2014.09.010
Kim W, Cho Y, Song M-K, Lim J-h, Jy K et al (2018) Effect of particulate matter 2.5 on gene expression profile and cell signaling in JEG-3 human placenta cells. Environ Toxicol 33:1123–1134. https://doi.org/10.1002/tox.22591
Kim HJ, Kim G, Lee J, Lee Y, Kim J-H (2022) Secretome of stem cells: roles of extracellular vesicles in diseases, stemness, differentiation, and reprogramming. Tissue Eng Regen Med 19:19–33. https://doi.org/10.1007/s13770-021-00406-4
Kloog I, Ridgway B, Koutrakis P, Coull BA, Schwartz JD (2013) Long- and short-term exposure to PM2.5 and mortality: using novel exposure models. Epidemiology 24:555–561. https://doi.org/10.1097/EDE.0b013e318294beaa
Liu J, Man R, Ma S, Li J, Wu Q et al (2015) Atmospheric levels and health risk of polycyclic aromatic hydrocarbons (PAHs) bound to PM2.5 in Guangzhou, China. Mar Pollut Bull 100:134–143. https://doi.org/10.1016/j.marpolbul.2015.09.014
Liu W, Xu Y, Liu W, Liu Q, Yu S et al (2018) Oxidative potential of ambient PM(2.5) in the coastal cities of the Bohai Sea, northern China: seasonal variation and source apportionment. Environ Pollut 236:514–528. https://doi.org/10.1016/j.envpol.2018.01.116
Lopez-Ornelas A, Mejia-Castillo T, Vergara P, Segovia J (2011) Lentiviral transfer of an inducible transgene expressing a soluble form of Gas1 causes glioma cell arrest, apoptosis and inhibits tumor growth. Cancer Gene Ther 18:87–99. https://doi.org/10.1038/cgt.2010.54
Lu P, Takai K, Weaver VM, Werb Z (2011) Extracellular matrix degradation and remodeling in development and disease. Cold Spring Harb Perspect Biol 3:a005058. https://doi.org/10.1101/cshperspect.a005058
Nava P, Laukoetter MG, Hopkins AM, Laur O, Gerner-Smidt K et al (2007) Desmoglein-2: a novel regulator of apoptosis in the intestinal epithelium. Mol Biol Cell 18:4565–4578. https://doi.org/10.1091/mbc.e07-05-0426
Nicin L, Wagner JUG, Luxan G, Dimmeler S (2022) Fibroblast-mediated intercellular crosstalk in the healthy and diseased heart. FEBS Lett 596:638–654. https://doi.org/10.1002/1873-3468.14234
Niida A, Hiroko T, Kasai M, Furukawa Y, Nakamura Y et al (2004) DKK1, a negative regulator of Wnt signaling, is a target of the beta-catenin/TCF pathway. Oncogene 23:8520–8526. https://doi.org/10.1038/sj.onc.1207892
Ning X, Wang Y, Zhu N, Li G, Sang N (2020) Risk assessment of the lipid metabolism-disrupting effects of nitro-PAHs. J Hazard Mater 396:122611. https://doi.org/10.1016/j.jhazmat.2020.122611
Niu B-Y, Li W-K, Li J-S, Hong Q-H, Khodahemmati S et al (2020) Effects of DNA damage and oxidative stress in human bronchial epithelial cells exposed to PM(2.5) from Beijing, China, in Winter. Int J Environ Res Public Health 17:4874. https://doi.org/10.3390/ijerph17134874
Ortiz A (2021) Extracellular vesicles in cancer progression. Semin Cancer Biol 76:139–142. https://doi.org/10.1016/j.semcancer.2021.05.032
Overmiller AM, Pierluissi JA, Wermuth PJ, Sauma S, Martinez-Outschoorn U et al (2017) Desmoglein 2 modulates extracellular vesicle release from squamous cell carcinoma keratinocytes. FASEB J 31:3412–3424. https://doi.org/10.1096/fj.201601138RR
Pei L, Zhao M, Xu J, Li A, Luo K et al (2019) Associations of ambient fine particulate matter and its constituents with serum complement C3 in a panel study of older adults in China. Environ Pollut 252:1019–1025. https://doi.org/10.1016/j.envpol.2019.05.096
Piao MJ, Ahn MJ, Kang KA, Ryu YS, Hyun YJ et al (2018) Particulate matter 2.5 damages skin cells by inducing oxidative stress, subcellular organelle dysfunction, and apoptosis. Arch Toxicol 92:2077–2091. https://doi.org/10.1007/s00204-018-2197-9
Popova NV, Juecker M (2022) The functional role of extracellular matrix proteins in cancer. Cancers (Basel) 14:238. https://doi.org/10.3390/cancers14010238
Reyes-Zárate E, Sánchez-Pérez Y, Gutiérrez-Ruiz MC, Chirino YI, Osornio-Vargas ÁR et al (2016) Atmospheric particulate matter (PM10) exposure-induced cell cycle arrest and apoptosis evasion through STAT3 activation via PKCζ and Src kinases in lung cells. Environ Pollut 214:646–656. https://doi.org/10.1016/j.envpol.2016.04.072
Rui W, Guan L, Zhang F, Zhang W, Ding W (2016) PM2.5-induced oxidative stress increases adhesion molecules expression in human endothelial cells through the ERK/AKT/NF-kappaB-dependent pathway. J Appl Toxicol 36:48–59. https://doi.org/10.1002/jat.3143
Shan N, Zhang X, Xiao X, Zhang H, Tong C et al (2015) Laminin alpha 4 (LAMA4) expression promotes trophoblast cell invasion, migration, and angiogenesis, and is lowered in preeclamptic placentas. Placenta 36:809–820. https://doi.org/10.1016/j.placenta.2015.04.008
Song X, Liu J, Geng N, Shan Y, Zhang B et al (2022) Multi-omics analysis to reveal disorders of cell metabolism and integrin signaling pathways induced by PM2.5. J Hazard Mater 424:127573. https://doi.org/10.1016/j.jhazmat.2021.127573
Strack R (2021) Revealing the secretome. Nat Methods 18:1273–1273. https://doi.org/10.1038/s41592-021-01320-2
Stuehler K (2020) The secrets of protein secretion: what are the key features of comparative secretomics? Expert Rev Proteomics 17:785–787. https://doi.org/10.1080/14789450.2020.1881890
Torres-Ramos Y, Montoya-Estrada A, Guzman-Grenfell A, Mancilla J, Cardenas B et al (2011) Urban PM2.5 induces ROS generation and RBC damage in COPD patients. Front Biosci (Elite edition) 3:808–817. https://doi.org/10.2741/e288
Wang W, Deng Z, Feng Y, Liao F, Zhou F et al (2017) PM2.5 induced apoptosis in endothelial cell through the activation of the p53-bax-caspase pathway. Chemosphere 177:135–143. https://doi.org/10.1016/j.chemosphere.2017.02.144
Weng Y, Sui Z, Shan Y, Jiang H, Zhou Y et al (2016) In-depth proteomic quantification of cell secretome in serum-containing conditioned medium. Anal Chem 88:4971–4978. https://doi.org/10.1021/acs.analchem.6b00910
Wu X, Zhu B, Zhou J, Bi Y, Xu S et al (2021) The epidemiological trends in the burden of lung cancer attributable to PM2.5 exposure in China. BMC Public Health 21:737. https://doi.org/10.1186/s12889-021-10765-1
Xia T, Shen Z, Cai J, Pan M, Sun C (2020) ColXV aggravates adipocyte apoptosis by facilitating abnormal extracellular matrix remodeling in mice. Int J Mol Sci 21:959. https://doi.org/10.3390/ijms21030959
Xu H, Jiao X, Wu Y, Li S, Cao L et al (2019a) Exosomes derived from PM2.5-treated lung cancer cells promote the growth of lung cancer via the Wnt3a/-catenin pathway. Oncol Rep 41:1180–1188. https://doi.org/10.3892/or.2018.6862
Xu M-X, Ge C-X, Qin Y-T, Gu T-T, Lou D-S et al (2019b) Prolonged PM2.5 exposure elevates risk of oxidative stress-driven nonalcoholic fatty liver disease by triggering increase of dyslipidemia. Free Radic Biol Med 130:542–556. https://doi.org/10.1016/j.freeradbiomed.2018.11.016
Xu D, Zhang Y, Sun Q, Wang X, Li T (2021) Long-term PM(2.5) exposure and survival among cardiovascular disease patients in Beijing, China. Environ Sci Pollut Res Int 28:47367–47374. https://doi.org/10.1007/s11356-021-14043-w
Xue Z, Li A, Zhang X, Yu W, Wang J et al (2019) iTRAQ based proteomic analysis of PM2.5 induced lung damage. RSC Adv 9:11707–11717. https://doi.org/10.1039/C9RA00252A
Yan Z, Wang J, Li J, Jiang N, Zhang R et al (2016) Oxidative stress and endocytosis are involved in upregulation of interleukin-8 expression in airway cells exposed to PM2.5. Environ Toxicol 31:1869–1878. https://doi.org/10.1002/tox.22188
Yang L, Liu G, Li X, Xia Z, Wang Y et al (2020) Small GTPase RAB6 deficiency promotes alveolar progenitor cell renewal and attenuates PM2.5-induced lung injury and fibrosis. Cell Death Dis 11:1–17. https://doi.org/10.1038/s41419-020-03027-2
Yang Z, Mahendran R, Yu P, Xu R, Yu W et al (2022) Health effects of long-term exposure to ambient PM2.5 in Asia-Pacific: a systematic review of cohort studies. Current Environmental. Health Rep. https://doi.org/10.1007/s40572-022-00344-w
Zarco N, Gonzalez-Ramirez R, Gonzalez RO, Segovia J (2012) GAS1 induces cell death through an intrinsic apoptotic pathway. Apoptosis 17:627–635. https://doi.org/10.1007/s10495-011-0696-8
Zhai X, Wang J, Sun J, Xin L (2022) PM2.5 induces inflammatory responses via oxidative stress-mediated mitophagy in human bronchial epithelial cells. Toxicol Res 11:195–205. https://doi.org/10.1093/toxres/tfac001
Zhang Z, He J, Shi T, Tang N, Zhang S et al (2019) Associations between polychlorinated dibenzo-dioxins and polychlorinated dibenzo-furans exposure and oxidatively generated damage to DNA and lipid. Chemosphere 227:237–246. https://doi.org/10.1016/j.chemosphere.2019.04.057
Zhang F, Yang B, Wang Y, Zhu J, Liu J et al (2020) Time- and dose-resolved proteome of PM(2.5)-exposure-induced lung injury and repair in rats. J Proteome Res 19:3162–3175. https://doi.org/10.1021/acs.jproteome.0c00155
Zhao C, Zhu L, Li R, Wang H, Cai Z (2019) Omics approach reveals metabolic disorders associated with the cytotoxicity of airborne particulate matter in human lung carcinoma cells. Environ Pollut 246:45–52. https://doi.org/10.1016/j.envpol.2018.11.108
Funding
This work was supported by National Key Research and Development Program of China (Grant 2017YFA0505003) and National Natural Science Foundation (Grants 21775149, 91543201, 21806165, and 21725506).
Author information
Authors and Affiliations
Contributions
Conceptualization: SZ, CJ, and ZL; methodology: SZ, SX, and WY; writing—original draft preparation: SZ, SX and WY; writing—review and editing: LJ, ZB and LZ; supervision: CJ, ZL and ZY; funding acquisition: ZL, SZ and SX. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Ethics approval and consent to participate
Not applicable
Consent for publication
Not applicable
Competing interests
The authors declare no competing interests.
Additional information
Responsible Editor: Mohamed M. Abdel-Daim
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary information
ESM 1
(XLSX 98 kb)
Rights and permissions
About this article
Cite this article
Sui, Z., Song, X., Wu, Y. et al. The cytotoxicity of PM2.5 and its effect on the secretome of normal human bronchial epithelial cells. Environ Sci Pollut Res 29, 75966–75977 (2022). https://doi.org/10.1007/s11356-022-20726-9
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s11356-022-20726-9