BackgroundThe hedgehog signaling system (HHS) plays an important role in the regulation of cell proliferation and differentiation during the embryonic phases. However, little is known about the involvement of HHS in the malignant transformation of cells. This study aimed to detect the role of HHS in the malignant transformation of human bronchial epithelial (16HBE) cells.

MethodsIn this study, two microfluidic chips were designed to investigate cigarette smoke extract (CSE)-induced malignant transformation of cells. Chip A contained a concentration gradient generator, while chip B had four cell chambers with a central channel. The 16HBE cells cultured in chip A were used to determine the optimal concentration of CSE for inducing malignant transformation. The 16HBE cells in chip B were cultured with 12.25% CSE (Group A), 12.25% CSE + 5 μmol/L cyclopamine (Group B), or normal complete medium as control for 8 months (Group C), to establish the in vitro lung inflammatory-cancer transformation model. The transformed cells were inoculated into 20 nude mice as cells alone (Group 1) or cells with cyclopamine (Group 2) for tumorigenesis testing. Expression of HHS proteins was detected by Western blot. Data were expressed as mean ± standard deviation. The t-test was used for paired samples, and the difference among groups was analyzed using a one-way analysis of variance.

ResultsThe optimal concentration of CSE was 12.25%. Expression of HHS proteins increased during the process of malignant transformation (Group B vs. Group A, F = 7.65, P < 0.05). After CSE exposure for 8 months, there were significant changes in cellular morphology, which allowed the transformed cells to grow into tumors in 40 days after being inoculated into nude mice. Cyclopamine could effectively depress the expression of HHS proteins (Group C vs. Group B, F = 6.47, P < 0.05) and prevent tumor growth in nude mice (Group 2 vs. Group 1, t = 31.59, P < 0.01).

ConclusionsThe activity of HHS is upregulated during the CSE-induced malignant transformation of 16HBE cells. Cyclopamine can effectively depress expression of HHS proteins in vitro and prevent tumor growth of the transformed cells in vivo.

Animals ; Cell Transformation, Neoplastic ; genetics ; metabolism ; Gene Expression Regulation, Neoplastic ; genetics ; physiology ; Hedgehog Proteins ; genetics ; metabolism ; Lab-On-A-Chip Devices ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Microfluidics ; Signal Transduction ; genetics ; physiology ; Smoke ; Smoking ; adverse effects

OBJECTIVE: We aimed to study the association between rs7525173, rs2236518, rs2493264 single nucleotide polymorphism (SNP) in the PRDM16 gene, smoking, alcohol exposures, and nonsyndromic cleft lip with or without cleft palate (NSCL/P). METHODS: A total of 157 case-parent trios were selected, and SNPs were genotyped by using ligase detection reaction (LDR) and direct sequencing methods. Transmission disequilibrium test (TDT) and linkage disequilibrium (LD) tests were con-ducted to analyze the data. A total of 1 710 patients with orofacial clefts and 956 healthy newborns were enrolled in the epidemiological survey. The smoking and drinking exposures of parents during early pregnancy were analyzed. RESULTS: The C allele at rs2236518 was over-transmitted for NSCPO (P<0.05). Statistical differences were observed among three factors, namely, maternal smoking, maternal passive smoking, and maternal drinking (P<0.05). CONCLUSIONS The rs2236518 at PRDM16 gene, maternal smoking, maternal passive smoking, and maternal drinking were closely related to the occurrence of NSCL/P.
Alcohol Drinking ; adverse effects ; Case-Control Studies ; Cleft Lip ; genetics ; Cleft Palate ; genetics ; DNA-Binding Proteins ; Environmental Exposure ; Female ; Genotype ; Humans ; Infant, Newborn ; Mothers ; Polymorphism, Single Nucleotide ; Pregnancy ; Smoking ; adverse effects ; Transcription Factors

Objective: To explore the SNP effects of patatin-like phospholipase domain which containing 3 (PNPLA3), transmembrane 6 superfamily member 2 (TM6SF2) gene, environmental effects of smoking, alcohol drinking and interaction between gene-gene, gene-environment and drinking-smoking on hepatitis B virus-associated hepatocellular carcinoma (HBV-HCC). Methods: We collected anticoagulant peripheral blood from patients of HBV-HCC, chronic hepatitis B (CHB), liver cirrhosis (LC) and from healthy controls to detect the single nucleotide polymorphism (SNP) of patatin-like phospholipase domain containing 3 (PNPLA3) gene loci rs738409 and transmembrane 6 superfamily member 2 (TM6SF2) gene loci rs58542926, using the flight mass spectrometry method. The optimal assignment value of gene polymorphisms was defined by using the online SNP stats. Hardy-Weinberg (H-W) balance was tested for SNP. Effects of the genetic and environmental factors to HBV-HCC were analyzed by using the multiple classification logistic regression method. The gene-gene, gene-smoking and alcohol drinking interaction effects were investigated by Fork-Life analysis and binary logistic regression methods. Results: The frequency distribution of CHB group rs738409 loci seemed not in conformity with the H-W balance (χ(2)=11.980, P<0.005). Two loci frequency distributions in the other groups were all in accordandce with the H-W balance. After adjusting for influences on age and sex and comparing to the healthy group, the rs58542926 mutation appeared as OR=1.659, 95%CI: 1.026-2.684, P=0.039, in the HBV-HCC group. When comparing to CHB group, the HBV-HCC group presented that drinking as OR=1.680, 95%CI: 1.121-2.519, P=0.012. When comparing to the LC group, the ORs of drinking and smoking were 1.539 (1.071-2.213) and 1.453 (1.005-2.099) respectively, in the HBV-HCC group. When comparing to the CHB+LC group, interactions between the HBV-HCC group were found rs738409 and rs58542926 on additive model OR=1.548 (U=1.885, P=0.029) and OR=1.658 (P=0.024) on logistic regression model while drinking was rs738409 on interaction additive model with OR=1.811(U=1.965, P=0.024). As for drinking and mutation of rs738409, the multiplication model of logistic regression showed no statistically significant differences. Interaction between smoking and drinking appeared as OR=1.756 (P<0.001) in the logistics regression multiplication model. Conclusions: Factors as mutation of TM6SF2, smoking and drinking all appeared as risk factors for HBV-HCC. Mutations of both PNPLA3 and TM6SF2, together with smoking and drinking all served as risk factors for HBV-HCC. However, the mutation of single PNPLA3 appeared as a protective factor on HBV-HCC.
Alcohol Drinking/adverse effects* ; Carcinoma, Hepatocellular/virology* ; Case-Control Studies ; Epistasis, Genetic ; Gene-Environment Interaction ; Genetic Predisposition to Disease ; Genotype ; Hepatitis B virus ; Hepatitis B, Chronic ; Humans ; Lipase/genetics* ; Liver Cirrhosis, Alcoholic/complications* ; Liver Neoplasms/virology* ; Membrane Proteins/genetics* ; Polymorphism, Single Nucleotide ; Smoking/adverse effects*

OBJECTIVETo investigate the cumulative effect regarding the family history of cardiovascular disease and smoking on ischemic stroke events in population with Mongolian ethnicity.

METHODSBased on data gathered from the baseline investigation, a 10-year prospective cohort follow-up project was conducted among 2 589 participants with Mongolian ethnicity. Ischemic stroke events were defined as the outcomes of the study. All the 2 589 participants were categorized into four subgroups: without family history of cardiovascular disease/nonsmokers, without family history of cardiovascular disease/smokers, with family history of cardiovascular disease/nonsmokers and with family history of cardiovascular disease/smokers, according to family history of cardiovascular disease and smoking status. Cumlative incidence rates of events among the four subgroups was described with Kaplan-Meier curves. Cox proportional hazards model was used to estimate the hazard ratios (HRs) and 95% confidence intervals (95%CI) of ischemic stroke events among the four subgroups.

RESULTSData from the Kaplan-Meier curves showed that the cumulative incidence rates of ischemic stroke were 1.17% (15/1 278), 3.83% (37/967), 5.70% (9/158) and 8.33% (15/180) for the groups of no family history of cardiovascular disease/nonsmokers, no family history of cardiovascular disease/smokers, with family history of cardiovascular disease/nonsmokers and with family history of cardiovascular disease/smokers, respectively. By cox proportional hazards model, after adjusting for age, male, drinking status, systolic and diastolic blood pressure, body mass index, fasting glucose, total cholesterol, triglycerides, LDL cholesterol factors, the HRs (95% CI) of ischemic stroke were 2.26 (1.19-4.28) and 2.45 (1.13-5.33) in the no family history of cardiovascular disease/smokers group, with family history of cardiovascular disease/smokers group when compared to the no family history of cardiovascular disease/nonsmokers group, respectively. The risk of ischemic stroke appeared the highest in the group with family history of cardiovascular disease/smokers (all P<0.05).

CONCLUSIONSmoking may increase the risk of ischemic stroke events among the population with family history of cardiovascular disease.

Alcohol Drinking ; Asian Continental Ancestry Group ; ethnology ; genetics ; Blood Glucose ; Blood Pressure ; Body Mass Index ; Cardiovascular Diseases ; ethnology ; genetics ; Cholesterol ; Cholesterol, LDL ; Genetic Predisposition to Disease ; Humans ; Incidence ; Male ; Mongolia ; epidemiology ; Population Surveillance ; Proportional Hazards Models ; Prospective Studies ; Risk Factors ; Smoking ; adverse effects ; epidemiology ; Stroke ; epidemiology ; genetics

OBJECTIVETo investigate the interaction on smoking and the lung cancer related genes miR-196a2 rs11614913, miR-146a rs2910164, miR-300 rs12894467, miR-26a-1 rs7372209, miR-27a rs895819 in Fujian Han population.

METHODSFrom January 2006 to January 2012, by using a hospital-based case-control study, 1 053 cases were pathologically diagnosed as primary lung cancer from the Department of Thoracic Surgery and 1 058 controls were randomly selected from the visiting relatives of patients and visiting people of Cangxia community health service of Fuzhou city according to match with age and genders. They were recruited for questionnaires survey and genotyping detection. Research objects of genders, height, weight, cultural degree, marital status, family history of cancer, lung disease history, smoking, drinking tea, drinking, and so on. After informed consent, we collected 5 ml fasting venous blood from every object, used MALDI-TOF-MS to analysis genotyping of polymorphic loci. Logistic regression model was constructed by using SNP as independent variable, and the multiple factors were constructed with different loci. The possible association between SNP and cigarette smoking was analyzed by using the crossover analysis. The relative excess risk of interaction (RERI) were used to analyze on smoking and SNP loci additive interaction of dominant and recessive genetic models.

RESULTSSmokers in case group who smoked P50(P25-P75)30.00 (0.00-56.00) packages in a year were higher than control group (0.00(0.00 - 20.48) pack years) (Z=14.57,P<0.001). Passive smoking index for non-smokers was 11.40(0.00-25.00), higher than the controls (0.00(0.00-13.11)) (Z=10.71,P<0.001). Site detection rate of rs11614913, rs2910164, rs12894467, rs7372209 and rs895819 in cases was 95.82%(1 009/1 053), 97.72%(1 029/1 053), 97.82% (1 030/1 053), 97.15% (1 023/1 053) and 96.01% (1 011/1 053) respectively. The controls were 98.11% (1 038/1 058), 98.96% (1 047/1 058), 98.30% (1 040/1 058), 98.68% (1 044/1 058) and 98.02% (1 037/1 058) respectively. rs11614913 dominant genetic model, TT genotype and smoking could increase the risk of primary lung cancer (OR=4.04, 95%CI: 2.67 -6.12). Recessive genetic model, CC genotype and smoking increased the incidence of primary lung cancer risk (OR=4.76, 95%CI: 3.16 -7.17). rs12894467 dominant genetic model, TT genotype and smoking could increase the risk (OR=2.98, 95%CI: 2.28 -3.90) in primary lung cancer. In recessive genetic model, CC genotype and smoking increased the incidence of primary lung cancer risk (OR=1.94, 95% CI: 1.10-3.43). Dominant genetic model of rs2910164, CC genotype and smoking could increase the risk (OR=2.18, 95% CI: 1.60 -2.98) in primary lung cancer. Recessive genetic model, GG genotype and smoking increased the incidence of primary lung cancer risk (OR=3.29, 95% CI: 2.16 -5.03). Especially rs12894467 dominant and recessive gene model and genders, smoking and there were combined effects(χ(2)=8.58, P=0.003; χ(2)=4.76, P=0.040).

CONCLUSIONRs11614913, rs12894467 and rs2910164 polymorphism were potentially associated with primary lung cancer in Fujian Han population.

Case-Control Studies ; China ; Genetic Predisposition to Disease ; Genotype ; Humans ; Lung Neoplasms ; genetics ; MicroRNAs ; genetics ; Polymorphism, Single Nucleotide ; Risk Factors ; Smoking ; adverse effects

The role of genetic polymorphisms of NAD(P)H:quinone oxidoreductase 1 (NQO1), which is known to be related to carcinogen metabolism and oxidative status, was evaluated for lung cancer development. The genotypes of two NQO1 polymorphisms, namely, IVS1-27C>G and Ex6+40C>T, were determined in 616 lung cancer cases and 616 lung cancer-free controls and haplotypes composed of the two polymorphisms were estimated. In the evaluation of the effect of the NQO1 genotypes or diplotypes, we did not find any significant association with lung cancer risk after adjusting for body mass index and smoking status. However, when we evaluated the effect of the NQO1 diplotypes for lung cancer risk in combination with smoking, smokers without the C-T/C-T diplotype showed a significantly increased risk of lung cancer compared with nonsmokers without the C-T/C-T diplotype (adjusted OR, 2.2; 95% CI, 1.67-3.02), and smokers with the C-T/C-T diplotype showed the highest OR of lung cancer (adjusted OR, 2.7; 95% CI, 1.78-4.21). Moreover, a trend test showed an additive interaction between smoking and the NQO1 C-T/C-T diplotype (P(trend) < 0.01). The additive effect of smoking and the NQO1 C-T/C-T diplotype was more apparent in squamous cell carcinoma, although this effect was statistically significant in all lung cancer cell types (all cell types, P(trend) < 0.05). This result suggests that haplotypes of the NQO1 gene play an important role in the development of lung cancer by interaction with smoking.
Aged ; Carcinoma, Non-Small-Cell Lung/epidemiology/*genetics ; Female ; Genetic Predisposition to Disease ; Haplotypes/genetics ; Humans ; Lung Neoplasms/epidemiology/*genetics ; Male ; Middle Aged ; NAD(P)H Dehydrogenase (Quinone)/*genetics ; Polymorphism, Single Nucleotide/genetics ; Risk ; Small Cell Lung Carcinoma/epidemiology/*genetics ; Smoking/*adverse effects

BACKGROUNDFound in inflammatory zone 1 (FIZZ1) protein increased in pulmonary epithelial cells and in limited amounts of other lung cells. FIZZ1 increased in murine model of smoke induced chronic obstructive pulmonary disease. However, the direct role of FIZZ1 produced by pulmonary epithelium stimulated with cigarette smoke extraction has not been determined. We examined the expression and function of FIZZ1 in rat lung epithelial L2 cells.

METHODSThe rat lung epithelial L2 cells (CCL 149) were exposed to cigarette smoke extraction, expression of FIZZ1 mRNA was investigated by RT-PCR. Levels of FIZZ1 protein were detected by Western blotting and laser confocal microscope. CCL 149 cells were treated with different concentrations and for different time of recombinant protein FIZZ1. After treatment, the expression levels of interleukin 8 (IL-8) were detected by enzyme-linked immunosorbent assay (ELISA).

RESULTSWhen CCL 149 cells were exposed to cigarette smoke extraction, FIZZ1 mRNA and protein levels expressed significantly higher than control group. Recombinant protein FIZZ1 promoted the expression of IL-8 in a dose and time dependent manner in a certain range.

CONCLUSIONSCigarette smoke extraction activates FIZZ1 at mRNA and protein levels in CCL 149 cells. Recombinant protein FIZZ1 induces the expression of IL-8 and may thus participate in the process of chronic obstructive pulmonary disease airway inflammation and airflow obstruction. Generally, immune cells such as macrophages, neutrophils and lymphocytes are unavoidably involved in airway inflammatory and immune responses to cigarette smoke, but it is still unclear whether their involvement in the pathogenesis of chronic obstructive pulmonary disease is based on the specific expression in lung epithelial cells of FIZZ1.

Animals ; Cell Line ; Disease Models, Animal ; Enzyme-Linked Immunosorbent Assay ; Epithelial Cells ; drug effects ; metabolism ; Interleukin-8 ; genetics ; metabolism ; Nerve Growth Factor ; genetics ; metabolism ; Pulmonary Disease, Chronic Obstructive ; genetics ; RNA, Messenger ; genetics ; Rats ; Smoking ; adverse effects

Cytochrome P-450 CYP1A1 ; genetics ; Genetic Predisposition to Disease ; Humans ; Logistic Models ; Pulmonary Disease, Chronic Obstructive ; complications ; Risk Factors ; Smoking ; adverse effects ; Venous Thromboembolism ; etiology ; genetics

OBJECTIVETo estimate the contribution of known identified risk factors to breast cancer incidence and mortality in China, and provide evidence to support the prevention and control of breast cancer for Chinese females.

METHODSWe calculated the proportion of breast cancer attributable to specific risk factors. Data on exposure prevalence were obtained from Meta-analyses and large-scale national surveys of representative samples of the Chinese population. Data on relative risks were obtained from Meta-analyses and large-scale prospective studies. Cancer mortality and incidence were taken from the Third National Death Survey and from cancer registries in China.

RESULTSThe first 5 risk factors of breast cancer in China were benign breast disease (RR = 2.62), family history of breast cancer (RR = 2.39), smoking (RR = 1.86), overweight (RR = 1.60) and age at menarche (RR = 1.54). The proportion of breast cancer deaths attributable to reproductive factors, lifestyle factors, benign breast disease, the use of external hormone and family history of breast cancer was 27.84%, 23.55%, 15.09%, 3.60% and 2.49%, respectively. The total population attributable fraction (PAF) was 55.95% for risk factors in our study. Overall, we estimated that 79 862 breast cancer cases and 22 456 deaths were attributed to the five risk factors in China in 2005.

CONCLUSIONSThe prevention and control of unhealthy lifestyle factors may significantly reduce the number and death of breast cancer in China.

Breast Diseases ; complications ; Breast Neoplasms ; epidemiology ; etiology ; genetics ; China ; epidemiology ; Female ; Genetic Predisposition to Disease ; Humans ; Menarche ; Meta-Analysis as Topic ; Overweight ; complications ; Risk Factors ; Smoking ; adverse effects

PURPOSE: This study was designed to determine the relationship of cigarette smoking to the frequency and qualitative differences among KRAS mutations in lung adenocarcinomas from Korean patients. MATERIALS AND METHODS: Detailed smoking histories were obtained from 200 consecutively enrolled patients with lung adenocarcinoma according to a standard protocol. EGFR (exons 18 to 21) and KRAS (codons 12/13) mutations were determined via direct-sequencing. RESULTS: The incidence of KRAS mutations was 8% (16 of 200) in patients with lung adenocarcinoma. KRAS mutations were found in 5.8% (7 of 120) of tumors from never-smokers, 15% (6 of 40) from former-smokers, and 7.5% (3 of 40) from current-smokers. The frequency of KRAS mutations did not differ significantly according to smoking history (p=0.435). Never-smokers were significantly more likely than former or current smokers to have a transition mutation (G-->A or C-->T) rather than a transversion mutation (G-->T or G-->C) that is known to be smoking-related (p=0.011). In a Cox regression model, the adjusted hazard ratios for the risk of progression with epidermal growth factor receptor-tyrosine kinase inhibitors (EGFR-TKIs) were 0.24 (95% CI, 0.14-0.42; p<0.001) for the EGFR mutation and 1.27 (95% CI, 0.58-2.79; p=0.537) for the KRAS mutation. CONCLUSION: Cigarette smoking did not influence the frequency of KRAS mutations in lung adenocarcinomas in Korean patients, but influenced qualitative differences in the KRAS mutations.
Adenocarcinoma/drug therapy/etiology/*genetics/pathology ; Adult ; Aged ; Aged, 80 and over ; Asian Continental Ancestry Group/genetics ; Female ; Humans ; Incidence ; Lung Neoplasms/drug therapy/etiology/*genetics/pathology ; Male ; Middle Aged ; *Mutation ; Mutation Rate ; Proportional Hazards Models ; Proto-Oncogene Proteins/*genetics ; Receptor, Epidermal Growth Factor/antagonists & inhibitors/genetics ; Smoking/adverse effects/*genetics ; Treatment Outcome ; ras Proteins/*genetics