An H&E slide of each array post construction was completed as a quality control measure. section and all gel bands quantified. After normalization, the percent splicing was calculated using the formula: spliced/(unspliced + spliced) 100. 1471-2407-8-229-S3.xls (27K) GUID:?56630EE5-50B5-420C-BAFA-6FA2014D75FE Abstract Background Although lung cancer is among the few malignancies for which we know the primary etiological agent (i.e., cigarette smoke), a precise understanding of the temporal sequence of events that drive tumor progression remains elusive. In addition to finding that cigarette smoke (CS) impacts the functioning of key pathways with significant roles in redox homeostasis, xenobiotic detoxification, cell cycle control, and endoplasmic reticulum (ER) functioning, our data highlighted a defensive role for the unfolded protein response (UPR) program. The UPR promotes cell survival by reducing the accumulation of aberrantly folded proteins through translation arrest, production of chaperone proteins, and increased degradation. Importance of the UPR in maintaining tissue health is evidenced by the fact that a chronic increase in defective protein structures plays a pathogenic role in diabetes, cardiovascular disease, Alzheimer’s and Parkinson’s syndromes, and cancer. Methods Gene and protein expression changes in CS exposed human cell cultures were monitored by high-density microarrays and Western blot analysis. Tissue arrays containing samples from 110 lung cancers were probed with antibodies to proteins of interest using immunohistochemistry. Results We show that: 1) CS induces ER stress and activates components of the UPR; 2) reactive species in CS that promote oxidative stress are primarily responsible for UPR activation; 3) CS exposure results in increased expression of several genes with significant roles in attenuating oxidative stress; and 4) several major UPR regulators are increased either in expression (i.e., BiP and eIF2) or phosphorylation (i.e., phospho-eIF2) in a majority of human lung cancers. Conclusion These data indicate that chronic ER stress and recruitment of one or more UPR effector arms upon exposure to CS may play a pivotal role in the etiology or progression of lung cancers, and that phospho-eIF2 and 3-O-(2-Aminoethyl)-25-hydroxyvitamin D3 BiP may have diagnostic and/or therapeutic potential. Furthermore, we speculate that upregulation of UPR regulators (in particular BiP) may provide a pro-survival advantage by increasing resistance to cytotoxic stresses such as hypoxia and chemotherapeutic drugs, and that UPR induction is a potential mechanism that could be attenuated or reversed resulting in a more efficacious treatment strategy for lung cancer. Background The long lag time between initiation of cigarette smoking and 3-O-(2-Aminoethyl)-25-hydroxyvitamin D3 cancer induction (estimated at 25 to 50 pack-years) [1,2] raises several fundamental questions concerning the eventual induction of tobacco-induced diseases for which there is little information: e.g., how does the lung adapt to the chronic assault of many decades of cigarette smoke (CS) exposure, what are the biological sequelae that occur in response to this adaptation and the continuous disruption of normal cellular homeostasis in the lung, and is this adaption a help or hindrance to lung cancer development? Our working hypothesis is Rabbit Polyclonal to CYB5R3 3-O-(2-Aminoethyl)-25-hydroxyvitamin D3 that a) tobacco-induced lung cancer is a complex process in which numerous pro-survival cellular systems have important contributory functions that both augment and modify the central role played by tobacco carcinogens and reactive oxygen/nitrogen species, and b) CS temporally shapes the course of lung carcinogenesis through chronic activation, and 3-O-(2-Aminoethyl)-25-hydroxyvitamin D3 eventual dysregulation, of normal cellular defense mechanisms. In our published [3-6] and unpublished studies using high-density oligonucleotide arrays and other techniques to define relevant CS-induced alterations in gene/protein expression and function in lung cells, we have attempted to place the impacted genes into biological context by developing a plausible mechanistic model.