Elsevier

Lung Cancer

Volume 71, Issue 2, February 2011, Pages 123-129
Lung Cancer

A combination of functional polymorphisms in the CASP8, MMP1, IL10 and SEPS1 genes affects risk of non-small cell lung cancer

https://doi.org/10.1016/j.lungcan.2010.04.016Get rights and content

Abstract

Exposure to tobacco smoke as well as environmental and occupational factors is the major cause of lung cancer. Non-small cell lung cancer (NSCLC) is the major histological type. Genes in pathways affecting inflammation, cellular stress and apoptosis are important, and the extent of inflammation in the lung could be affected by polymorphisms modifying these responses. In the present study we have investigated whether a combination of potential functional polymorphisms in genes related to inflammation may modulate risk of NSCLC. Eleven functional polymorphisms in nine genes were analyzed for association with risk of NSCLC in 882 subjects from the Norwegian population. The results showed that individuals carrying combination of three functional polymorphisms in the caspase-8, matrix metalloproteinase-1, seleno-protein S1, and interleukin-10 genes had two-fold increased risk of NSCLC (OR 2.06 (95% CI, 1.19–3.47) whereas individuals with four risk genotypes had 4.62-fold increased risk (OR 4.62, 95% CI, 1.69–12.63). These results highlight the need to investigate the combinatory effects of multiple SNPs in the carcinogenesis of the lung.

Introduction

Exposure to tobacco smoke as well as environmental and occupational factors is major cause of lung cancer [1]. Enhanced inflammation due to environmental and intrinsic factors is an important co-factor in promotion and progression of lung cancer. Major genes modulating the level of inflammation in chemical carcinogenesis include interleukins 1 (IL-1) and 6 (IL-6), tumor necrosis factor alpha (TNF-α), and the transcription factor nuclear factor kappa B (NF-κB). Cigarette smoke also induces an endoplasmic reticulum (ER) stress response in the lung [2]. Several genes including Selenoprotein S (SEPS1) have been implicated in regulation of the ER-stress response.

Studies suggest that inflammatory reactions in the lung could be affected by polymorphisms in the genes regulating these responses. For example, polymorphisms in the TNF-α (G−308A, G+488A) and NF-κB (Ins-94Del) genes have been shown to modulate risk of several cancers [3], [4], [5], [6]. We have recently demonstrated that regulatory polymorphisms in the IL1B gene were correlated with increased risk of NSCLC and higher expression of IL1B mRNA in the lung [7]. In vitro studies using human lung epithelial cells showed that the functional polymorphism in the IL1B gene promoter (IL1B-31 T/C) had increased expressional activity after exposure to the lung carcinogen benzo(a)pyrene [8]. Among others, SEPS1 has been identified as an ER protein participating in processing and removal of misfolded proteins from the ER to the cytosol where they are destroyed by the proteasome in an ubiquitin-dependent manner [9]. Recently, the SEPS1 G-105A promoter SNP was identified and shown to influence the efficiency of removal of the misfolded proteins from the ER, resulting in increased ER-stress and elevated circulatory levels of the pro-inflammatory cytokines, particularly IL1β [9]. An interaction between the SEPS1 -105 SNP and the IL1B promoter SNPs towards an increased risk of rheumatid arthritis has been suggested as evidence for epistasis between the IL1B and SEPS1 genes [10].

The cysteine-dependent aspartate-specific protease-8 (CASP-8) is involved in apoptotic signaling as well as inflammation [11]. Apoptotic signals can lead to activation and cleavage of pro-caspase 8 into active caspase 8, thereby inducing apoptosis [12]. Caspase-8 may also trigger inflammation [13]. A functional six nucleotide insertion–deletion polymorphism (-652 ins/del) in the CASP8 gene promoter was recently described and reported to be associated with susceptibility to multiple cancers [14]. Yet another caspase protein, Caspase-1 (CASP-1) may mediate cytokine production caused by tobacco smoke. CASP-1 enzyme is essential for the cleavage of pro-IL1β protein into its active and mature form [15]. A polymorphism in the CASP1 gene (G+5455A) may affect the enzymatic activity of the CASP1 enzyme towards pro-IL1β and hence formation of the active IL1β protein [16].

The inflammation-related molecules such as the matrix metalloproteinase 1 (MMP-1) are associated with degradation of the extracellular matrix (ECM), tissue remodeling and alteration of cellular signals in multiple cancer types [17]. MMP-1 may also be involved in the migration of immune cells from the bloodstream to sites of inflammation. A functional insertion/deletion polymorphism in the -1607 promoter region (-1607 2G/1G) of the MMP1 gene has been shown to affect expression levels of the gene [18].

Interleukin-10 (IL-10) is an important immune regulatory cytokine, mostly with anti-inflammatory functions [19], [20]. The implication of IL-10 in lung cancer has previously been studied, but its significance is still not clear [21], [22]. Three promoter SNPs at positions -1082, -819 and -592, forming a specific haplotype, have been associated with decreased expression of IL10 gene [23].

Studies indicate that Toll-like receptor 4 (TLR-4) signaling contributes to environmentally induced airway inflammation [24]. The A-2026G polymorphism in the regulatory region of the TLR4 gene has been shown to be functional and has been associated with several diseases [25], [26], [27]. Another gene involved in the TLR4 signaling is the CD14 which is a receptor for bacterial wall components such as LPS and is a susceptibility locus for asthma and may be a critical factor in lung inflammation as well [28]). The functional C-260T polymorphism in the CD14 gene has been shown to affect expression of the gene [29].

We hypothesized that functional polymorphisms that are individually associated with a weak to moderate risk of lung cancer, may have additive effects leading to a much higher lung cancer risk. We examined the association between NSCLC and 11 functionally verified polymorphisms in genes related to lung inflammation, cellular stress and apoptosis. Our results suggest that a significant increase in the risk of NSCLC may require a combination of at least three functional polymorphisms. Specifically, we found that a combination of functional polymorphisms in the CASP8, MMP1, IL10 and SEPS1 genes increased risk of NSCLC to more than four-fold. The odds ratios were tested and further confirmed using statistical tools False-Positive Report Probability (FPRP) and Bayesian False Discovery Probability (BFDP) tests [30], [31] developed to correct for multiple testing errors in association studies.

In view of the significance of the TP53 gene in lung carcinogenesis, we also examined the genotypes in relation to TP53 mutations in the lung tumors available from a subset of lung cancer patients.

Section snippets

Study population

The characteristics of lung cancer patients and healthy controls included in the present study are summarized in Table 1. The details of the study population are recently published by Landvik et al. [7]. Briefly, 455 lung cancer patients were admitted for surgery at the university hospitals in Oslo or Bergen between 1986 and 2001. Diagnosis of lung cancer was ascertained by reviewing histological slides from tumor tissue and 442 non-small cell lung cancer (NSCLC) cases were enrolled in the

Results

The characteristics of NSCLC patients and healthy controls are shown in Table 1. Eleven polymorphisms in nine genes (Table 2), involved in inflammation, cellular stress and apoptosis were analyzed for association with risk of NSCLC. The polymorphisms were selected on the basis of confirmed functionality [3], [9], [14], [16], [18], [23], [25], [29], [37], [38]. Genotypes were obtained for >92% of cases and controls for all eleven polymorphisms as shown in online supplementary Tables 1 and 2 (

Discussion

In this study we have examined the risk of NSCLC associated with eleven functional polymorphisms in nine genes. We investigated further the interaction between four of the polymorphisms in CASP8, MMP1, SEPS1 and IL10 genes. The results showed a step-wise increase in risk of NSCLC with increasing number of risk genotypes.

IL-10 is an important immunoregulatory cytokine with anti-inflammatory properties and capacity to inhibit pro-inflammatory cytokines including IL-1β, IL-6 and TNF-α [39] and is

Conflict of interest

The authors declare no conflicts of interest.

Acknowledgments

The authors gratefully acknowledge collaboration of Dr. Lodve Stangeland, Haukeland University Hospital, Bergen, and Dr. Anne Naalsund, National University Hospital, Oslo for recruiting the lung cancer patients. We are grateful to Ms. Elín Einarsdóttir Thornér, Ms. Tove Andreassen and Mr. Erik Eide for excellent technical assistance. The National Health Screening Service (Norway) is acknowledged for collecting and providing biological materials and demographic data for the controls.

Funding:

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