Comparison of Proteomic and Transcriptomic Profiles in the Bronchial Airway Epithelium of Current and Never Smokers

University of Pittsburgh, United States of America
PLoS ONE (Impact Factor: 3.53). 02/2009; 4(4):e5043. DOI: 10.1371/journal.pone.0005043
Source: PubMed

ABSTRACT Although prior studies have demonstrated a smoking-induced field of molecular injury throughout the lung and airway, the impact of smoking on the airway epithelial proteome and its relationship to smoking-related changes in the airway transcriptome are unclear.
Airway epithelial cells were obtained from never (n = 5) and current (n = 5) smokers by brushing the mainstem bronchus. Proteins were separated by one dimensional polyacrylamide gel electrophoresis (1D-PAGE). After in-gel digestion, tryptic peptides were processed via liquid chromatography/ tandem mass spectrometry (LC-MS/MS) and proteins identified. RNA from the same samples was hybridized to HG-U133A microarrays. Protein detection was compared to RNA expression in the current study and a previously published airway dataset. The functional properties of many of the 197 proteins detected in a majority of never smokers were similar to those observed in the never smoker airway transcriptome. LC-MS/MS identified 23 proteins that differed between never and current smokers. Western blotting confirmed the smoking-related changes of PLUNC, P4HB1, and uteroglobin protein levels. Many of the proteins differentially detected between never and current smokers were also altered at the level of gene expression in this cohort and the prior airway transcriptome study. There was a strong association between protein detection and expression of its corresponding transcript within the same sample, with 86% of the proteins detected by LC-MS/MS having a detectable corresponding probeset by microarray in the same sample. Forty-one proteins identified by LC-MS/MS lacked detectable expression of a corresponding transcript and were detected in <or=5% of airway samples from a previously published dataset.
1D-PAGE coupled with LC-MS/MS effectively profiled the airway epithelium proteome and identified proteins expressed at different levels as a result of cigarette smoke exposure. While there was a strong correlation between protein and transcript detection within the same sample, we also identified proteins whose corresponding transcripts were not detected by microarray. This noninvasive approach to proteomic profiling of airway epithelium may provide additional insights into the field of injury induced by tobacco exposure.

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Available from: James Flanigon, Jul 20, 2015
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    • "Prostate cancer (correlation with malignity degree) [176] [177] [178] Erp57, Erp72, PDI P, I Chronic lymphocytic leukemia (with prolonged survival) and acute promyelocytic leukemia (methotrexate treatment) [179] [180] PDI, Erp57 P, I, G Breast cancer (in metastatic node vs primary tumors) [181] [182] [183] [184] [185] P5 I, G Hypoxia (in head and neck squamous cell line carcinoma) [186] Erp57 I Esophageal carcinoma (cells) [187] PDI P Oral squamous carcinoma (cells) [188] Erp57 P Gastric cancer [189] Erp46, Erp57 P, I Colorectal cancer (increases with 5-fluorouracil treatment) [190] [191] ER60 P, I Liver cancer (pheophorbide and photodynamic therapy in HepG2 cells) [192] Erp72, Erp57 P, I Hepatocellular carcinoma (increases in high vs low metastasis potential) [193] [194] Erp57, PDIp P Pancreatic carcinoma (increases in 5-aza-2′-deoxycytidine treatment) [195] [196] [197] PDI, Erp57, P5 P, I HeLa cells (resistance to aplidin) [198] PDI P, I H-RasV12-transformed p38α-deficient mouse embryo fibroblasts [199] Neurologic PDI, Erp57 P Hemiparkinson (in rat model) [200] Erp57 P Attention-deficit hyperactivity disorder (in midbrain and striatum, rat model) [201] PDI I Alzheimer disease (in temporal cortex brain) [202] PDI and Erp57 P, I Amyotrophic lateral sclerosis (in spinal cords from ALS patients vs controls, or G93A-SOD1 rat/mouse vs wild type) [203] [204] [205] Erp57 P Perinatal asphyxia (in hippocampus) [206] PDI I, G Brain ischemia (in dentate gyrus) [207] Erp57 P, I Sporadic Creutzfeldt-Jakob disease (in cerebellum) [208] Erp57 P Major depression (with paroxetine treatment) [209] PDI G Brain injury (with p-hydroxybenzyl alcohol treatment) [210] PDI P Tuberous sclerosis (HeLa cells transfected with TSC1/2 tumor suppressor gene) [211] Cardiovascular Erp57 P Endothelial function (improvement with salvianolic acid) [212] PDI P Acute coronary syndrome (in monocytes from atorvastatin-treated patients) [213] PDI I, G Ischemic heart disease (peri-infarct area vs remote area; murine HL-1 cells subjected to hypoxia vs control cells; MCP-1 Tg mice hearts vs wild type) [214] [215] PDI, Erp57, Erp44 P, I Preeclampsia (in placentae and trophoblasts) [216] [217] [218] Inflammation Erp46 I, G Varioliform gastritis [219] PDI P Antiphospholipid syndrome (with fluvastatin treatment) [220] Erp72 P Acute pancreatitis (in acinar cells) [221] Endocrine/metabolic Erp57 P, G Hepatosteatosis (in hepatic cells) [222] Erp57 P, G Obesity (subcutaneous adipose tissue in patients; white adipose tissue in leptin-treated ob/ob mice) [223] [224] PDI P, I Dyslipidemia (in fatty liver from high-fructose-fed hamsters) [225] Erp57 P, I Insulin resistance (in fat of obese subjects) [226] PDI, Erp57, P5 P, I Diabetes (in liver microsomes of diabetic rats; in islets exposed to high glucose) [227] [228] PDI P Insulin resistance and dyslipidemia (in hepatocytes) [229] Erp29, Erp72 I Congenital hypothyroid goiter (in thyroid tissues) [230] Others PDI P Hepatotoxicity (in carbon tetrachloride-induced liver injury) [231] ERP57 P, I Irritable bowel syndrome (in rat colon) [232] PDI P Helicobacter pylori infection (in gastric epithelial cells) [233] PDI P, I, G Smoker's lung [234] [235] Erp57 P, I Environmental contaminant (in nonylphenol-exposed Sertoli cells) [236] Erp57 P Periodontitis (in fibroblasts) [237] Erp57 P, I Dehydration (in rat hypothalamic supraoptic nucleus) [238] PDI, Erp57 P Aging (in colon epithelia and dermal fibroblasts) [239] [240] PDI I Pregnancy/embryo implantation (in decidualized endometrial lysates) [241] PDI P Episodic hypoxia (in rat kidney) [242] P, proteome analysis; I, immunodetection; G, gene expression analysis. "
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