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BACKGROUND: The impacts of chronic airway diseases on coronavirus disease 2019 (COVID-19) are far from understood. OBJECTIVE: To explore the influence of asthma and chronic obstructive pulmonary disease (COPD) comorbidity on disease expression and outcomes, and the potential underlying mechanisms in COVID-19 patients. METHODS: A total of 961 hospitalized COVID-19 patients with a definite clinical outcome (death or discharge) were retrospectively enrolled. Demographic and clinical information were extracted from the medical records. Lung tissue sections from patients suffering from lung cancer were used for immunohistochemistry study of angiotensin-converting enzyme II (ACE2) expression. BEAS-2B cell line was stimulated with various cytokines. RESULTS: In this cohort, 21 subjects (2.2%) had COPD and 22 (2.3%) had asthma. After adjusting for confounding factors, COPD patients had higher risk of developing severe illness (OR: 23.433; 95% CI 1.525-360.135; P < .01) and acute respiratory distress syndrome (OR: 19.762; 95% CI 1.461-267.369; P = .025) than asthmatics. COPD patients, particularly those with severe COVID-19, had lower counts of CD4+ T and CD8+ T cells and B cells and higher levels of TNF-α, IL-2 receptor, IL-10, IL-8, and IL-6 than asthmatics. COPD patients had increased, whereas asthmatics had decreased ACE2 protein expression in lower airways, compared with that in control subjects without asthma and COPD. IL-4 and IL-13 downregulated, but TNF-α, IL-12, and IL-17A upregulated ACE2 expression in BEAS-2B cells. CONCLUSION: Patients with asthma and COPD likely have different risk of severe COVID-19, which may be associated with different ACE2 expression.
Assuntos
Asma/epidemiologia , COVID-19/complicações , Doença Pulmonar Obstrutiva Crônica/epidemiologia , Idoso , Enzima de Conversão de Angiotensina 2/biossíntese , Asma/imunologia , Asma/metabolismo , COVID-19/imunologia , Comorbidade , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Prevalência , Doença Pulmonar Obstrutiva Crônica/imunologia , Doença Pulmonar Obstrutiva Crônica/metabolismo , SARS-CoV-2Assuntos
Enzima de Conversão de Angiotensina 2/genética , COVID-19/enzimologia , Rinite Alérgica/enzimologia , SARS-CoV-2 , Adulto , Idoso , COVID-19/etiologia , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Estudos Retrospectivos , Rinite Alérgica/complicações , Rinite Alérgica/epidemiologiaRESUMO
Cigarette smoke is associated with the development of several diseases, such as chronic obstructive pulmonary disease (COPD). The purpose of this study was to investigate genotoxicity and heat shock protein 70 (Hsp70) in human airway smooth muscle cells (HASMCs) exposed to cigarette smoke extract (CSE). HASMCs was exposed to CSE with different doses for 24 h. The level of 8-hydroxydeoxyguanosine (8-OHdG) was determined by using HPLC-ECD, the DNA damage was analyzed by using comet assay, and apoptosis was examined by using Annexin-FITC/PI staining. The production of Hsp70 after CSE stimulation was tested. Results indicated that CSE significantly increased the level of 8-OHdG, DNA damage and cell apoptosis, and reduced the production of Hsp70. In particular, levels of Hsp70 were inversely correlated with 8-OHdG, DNA damage and cell apoptosis. It was concluded that cigarette smoke induced genotoxicity and decreased the production of cell protective protein Hsp70, which may contribute to the development of some airway diseases.
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Dano ao DNA , Proteínas de Choque Térmico HSP70/metabolismo , Pulmão/efeitos dos fármacos , Miócitos de Músculo Liso/efeitos dos fármacos , Nicotiana/toxicidade , Fumaça/efeitos adversos , 8-Hidroxi-2'-Desoxiguanosina , Apoptose , Desoxiguanosina/análogos & derivados , Desoxiguanosina/metabolismo , Proteínas de Choque Térmico HSP70/genética , Humanos , Pulmão/citologia , Miócitos de Músculo Liso/metabolismo , Células Tumorais CultivadasRESUMO
In order to study whether cysteine-rich 61 protein (cyr61) is involved in the pathogenesis of asthma and its relation to airway inflammation, the effect of dexamethasone (Dxm) on the expression of cyr61 in the lung tissues of asthmatic mice was investigated. Forty BALB/c mice were divided into asthma group (n=15), control group (n=10) and Dxm group (n=15). The asthma group was sensitized and challenged by ovalbumin (OVA). The mice in Dxm group were intraperitoneally administered with Dxm after OVA challenge. The expression of cyr61 in the lung tissues was detected by using immunohistochemistry, and that of eotaxin protein in the bronchoalveolar lavage fluid (BALF) by using enzyme-linked immunosorbent assay (ELISA). The number of inflammatory cells in BALF was also analyzed. The results showed that the cyr61 expression was highest in asthma group (P<0.05), followed by Dxm group (P<0.05) and control group. The cyr61 had a positive correlation with the total nucleated cells (r=0.867, P<0.05), especially eosinophils (r=0.856, P<0.05), and eotaxin level (r=0.983, P<0.05) in the BALF. Our findings suggested that cyr61 is expressed in airway epithelial cells and has a positive correlation with eotaxin and number of airway infiltrating eosinophils.
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Asma/tratamento farmacológico , Proteína Rica em Cisteína 61/biossíntese , Dexametasona/farmacologia , Células Epiteliais/efeitos dos fármacos , Animais , Anti-Inflamatórios/administração & dosagem , Anti-Inflamatórios/farmacologia , Asma/induzido quimicamente , Asma/metabolismo , Líquido da Lavagem Broncoalveolar/química , Líquido da Lavagem Broncoalveolar/citologia , Quimiocinas CC/metabolismo , Dexametasona/administração & dosagem , Ensaio de Imunoadsorção Enzimática , Células Epiteliais/metabolismo , Células Epiteliais/patologia , Feminino , Imuno-Histoquímica , Injeções Intraperitoneais , Contagem de Leucócitos , Pulmão/metabolismo , Pulmão/patologia , Camundongos , Camundongos Endogâmicos BALB C , Neutrófilos/efeitos dos fármacos , Neutrófilos/patologia , OvalbuminaRESUMO
BACKGROUND AND OBJECTIVES: The majority of coronavirus disease 2019 (COVID-19) cases are nonsevere, but severe cases have high mortality and need early detection and treatment. We aimed to develop a nomogram to predict the disease progression of nonsevere COVID-19 based on simple data that can be easily obtained even in primary medical institutions. METHODS: In this retrospective, multicenter cohort study, we extracted data from initial simple medical evaluations of 495 COVID-19 patients randomized (2:1) into a development cohort and a validation cohort. The progression of nonsevere COVID-19 was recorded as the primary outcome. We built a nomogram with the development cohort and tested its performance in the validation cohort. RESULTS: The nomogram was developed with the nine factors included in the final model. The area under the curve (AUC) of the nomogram scoring system for predicting the progression of nonsevere COVID-19 into severe COVID-19 was 0.875 and 0.821 in the development cohort and validation cohort, respectively. The nomogram achieved a good concordance index for predicting the progression of nonsevere COVID-19 cases in the development and validation cohorts (concordance index of 0.875 in the development cohort and 0.821 in the validation cohort) and had well-fitted calibration curves showing good agreement between the estimates and the actual endpoint events. CONCLUSIONS: The proposed nomogram built with a simplified index might help to predict the progression of nonsevere COVID-19; thus, COVID-19 with a high risk of disease progression could be identified in time, allowing an appropriate therapeutic choice according to the potential disease severity.
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Abnormal hypertrophy and hyperplasia of airway smooth muscle cells play an important role in airway remodeling in chronic asthma. The authors' previous studies have indicated that protein kinase C alpha (PKC alpha) is involved in the proliferation of passively sensitized human airway smooth muscle cells (HASMCs). However, the underlying mechanisms remain unknown. Here, the authors examined the possible role of the alpha isoform of PKC in the control of cyclin D1 expression, using HASMCs passively sensitized on human atopic asthmatic serum as a model system. Cultured HASMCs were passively sensitized with serum from atopic asthmatic patients. Cell proliferation was measured by [(3)H]thymidine incorporation and an MTT assay. Cell cycle status was analyzed by flow cytometry. The mRNA and protein expression profiles of cyclin D1 were measured by reverse transcriptase-polymerase chain reaction (RT-PCR) and Western blotting, respectively. Furthermore, the authors assessed the role of cyclin D1 in PKC alpha-induced HASMC proliferation by transfection with a recombinant cyclin D1 antisense construct. The activation of PKC alpha with phorbol myristate acetate (PMA), a PKC activator, up-regulated cyclin D1 expression and increased the proliferation of passively sensitized HASMCs. This effect was significantly decreased by specific inhibition of PKC alpha with Go6976. In addition, the authors showed that transfection with antisense cyclin D1 abolished PMA-induced G1/S progression and HASMC proliferation. These results demonstrate that PKC alpha promotes the proliferation of HASMCs sensitized with atopic asthmatic serum via up-regulation of cyclin D1 expression.
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Asma/enzimologia , Brônquios/enzimologia , Proliferação de Células , Ciclina D1/metabolismo , Miócitos de Músculo Liso/fisiologia , Proteína Quinase C-alfa/metabolismo , Adulto , Asma/patologia , Brônquios/patologia , Carbazóis , Células Cultivadas , Feminino , Humanos , Isoenzimas , Masculino , Pessoa de Meia-Idade , Soro , Acetato de Tetradecanoilforbol , Regulação para Cima , Adulto JovemRESUMO
The authors have retracted the article [Hsa-miR-623 suppresses tumor progression in human lung adenocarcinoma, Cell Death & Disease volume 7, page e2388 (2016), doi 10.1038/cddis.2016.260] because it has recently come to their attention that the A549 cells used in this research were contaminated with Hela cells, which may have altered the outcome of their experiment. The conclusions of this article are therefore unreliable. All authors agree to this retraction.
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AIM: To determine whether protein kinase C (PKC) has any effect on the expression of cyclinD1, a key regulator of growth control and G1/S transition, and to investigate the underlying molecular mechanisms of PKC involving the remodeling of the asthmatic airway smooth muscle (ASM). METHODS: The treatment of synchronized ASM cells from asthmatic rats with PKC-specific agonist phorbol 12-myristate 13-acetate (PMA) and antagonist 2-{1-[3-(amidinothio) propyl]-1Hindol-3-yl}-3-(1-methylindol-3-yl) maleimide methanesulfonate salt (Ro31-8220) was followed by the proliferation assay. PKCalpha and cyclinD1 expressions in ASM cells (ASMC) were detected by RT-PCR and Western blotting. The relation between PKCalpha and cyclinD1 was assessed by linear regression analysis. The effect of the construct recombinant plasmid pcDNA3.1-antisense cyclinD1 (pcDNA3.1-ascyclinD1) on the proliferation of ASMC was found to be induced by PMA. RESULTS: The data showed phorbol ester-dependent PKCalpha promoted the proliferation of ASMC. The closely-positive correlation existed between the expression of PKCalpha and cyclinD1 at the transcriptional (r=0.821, P<0.01) and translational (r=0.940, P<0.01) levels. pcDNA3.1-ascyclinD1 could inhibit the proliferation of ASMC. pcDNA3.1-ascyclinD1 almost completely attenuated the PMA-induced proliferation effect as Ro31-8220+pcDNA3.1. CONCLUSION: The proliferation of ASMC by PKC might by regulated by the cyclinD1 expression in asthmatic rats.
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Asma/patologia , Proliferação de Células/efeitos dos fármacos , Ciclina D1/biossíntese , Miócitos de Músculo Liso/efeitos dos fármacos , Miócitos de Músculo Liso/patologia , Proteína Quinase C/fisiologia , Animais , Células Cultivadas , Ciclina D1/genética , Impressões Digitais de DNA , Regulação da Expressão Gênica , Masculino , Plasmídeos/genética , Antígeno Nuclear de Célula em Proliferação/metabolismo , Ratos , Ratos Endogâmicos BNRESUMO
BACKGROUND: Airway smooth muscle (ASM) is suspected to be a determining factor in the structural change of asthma. However, the role of protein kinase C alpha (PKCalpha) and cyclin D1 involved in the dysfunction of ASM leading to asthmatic symptoms is not clear. In this study, the central role of PKCalpha and cyclin D1 in ASM proliferation in asthmatic rats was explored. METHODS: Thirty-six pathogen-free male Brown Norway (BN) rats were randomly divided into 2 groups: control groups (group N1, N2 and N3) and asthmatic groups (group A1, A2, and A3). Groups A1, A2 and A3 were challenged with ovalbumin (OA) for 2 weeks, 4 weeks and 8 weeks respectively. Control animals were exposed to an aerosolized sterile phosphate buffered saline (PBS). The ASM mass and nucleus numbers were studied to estimate the degree of airway remodeling by the hematoxylin-eosin staining method. PKCalpha and cyclin D1 expression in the ASM cells was detected by reverse transcription polymerase chain reaction (RT-PCR) and immunohistochemistry. The relation between PKCalpha and cyclin D1 was assessed by linear regression analysis. PKC agonist phorbol 12-myristate 13-acetate (PMA), PKC inhibitor Ro31-8220 and an antisense oligonucleotide against cyclin D1 (ASOND) were used to treat ASM cells (ASMCs) obtained from the 2 weeks asthmatic rats. The cyclin D1 protein expression level was detected by Western blotting. RESULTS: Compared with the control group, the PKCalpha and cyclin D1 mRNA levels were increased in the asthmatic group. Similar to RT-PCR results, immunohistochemistry analysis for PKCalpha and cyclin D1 expression revealed an increased production in ASMCs after allergen treatment for 2, 4 and 8 weeks compared with the respective control groups. No difference in expression of PKCalpha and cyclin D1 in ASM were found in the 2, 4 or 8 weeks asthmatic rats. There were significant positive correlations between PKCalpha and cyclin D1 expression, both transcriptionally (r = 0.944, P < 0.01) and translationally (r = 0.826, P < 0.01), in ASM. The content of cyclin D1 in asthmatic ASMCs increased after being stimulated by PMA, and decreased when induced by Ro31-8220. ASOND targeting for cyclin D1 lowered the expression of cyclin D1 induced by PMA. CONCLUSIONS: Increased expression of PKCalpha and cyclin D1 in ASM along with smooth muscle structure changes might implicate PKCalpha and cyclin D1 participation in the proliferation of ASM and contribute to the pathogenesis of asthma after repeated allergen exposure in rats. The results suggested that cyclin D1 might be downstream of PKC signal transduction pathway.
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Asma/patologia , Ciclina D1/fisiologia , Pulmão/patologia , Miócitos de Músculo Liso/patologia , Proteína Quinase C-alfa/fisiologia , Animais , Proliferação de Células , Ciclina D1/genética , Masculino , Proteína Quinase C-alfa/genética , RNA Mensageiro/análise , Ratos , Ratos Endogâmicos BNRESUMO
This study is to investigate the expression of CyclinD1 in asthmatic rats and construct expression plasmids of sense and antisense CyclinD1 gene and transfect them to asthmatic airway smooth muscle cell to study the effects of CyclinD1 on the proliferation of airway smooth muscle cells in asthmatic rats. CyclinD1 cDNA was obtained by RT-PCR of total RNA extracted from the airway smooth muscle in asthmatic rats. The sequence was inserted into eukaryotic expression vector pcDNA3.1 (+) to recombinate the sense and antisense pcDNA3.1-CyclinD1 eukaryotic expression vector. The two recombinations and vector were then separately transfected into airway smooth muscle cell in asthmatic rats by using liposome. The expression level of CyclinD1 was certificated by Western blotting analysis. The proliferations of ASMCs isolated from asthmatic rats were examined with cell cycle analysis, MTT colorimetric assay and proliferating cell nuclear antigen (PCNA) immunocytochemical staining. Results showed (1) Compared with control group, the content of CyclinD1 was significantly increased; (2) It was comformed by restriction endonucleasa digestion and DNA sequence analysis that the expression plasmid of sense and antisense CyclinD1 were successfully recombinated. There was significant change of CyclinD1 expression between vector and sense CyclinD1 transfected cells, and the expression level of CyclinD1 in ASMC transfected with antisense CyclinD1 was lower than that in vector transfected cells (P <0.01); (3) In the asthmatic groups, compared with the vecter group, the percentage of S + G2M phase, absorbance A value of MTT and the expression rate of PCNA protein in ASMC transfected with pcDNA3. 1-CyclinD1 vector significantly increased. The values decreased remarkably in the pcDNA3,1-as CyclinD1 group. Statistical analysis revealed that there were significant differences in these indicators of cell proliferation in three groups (P <0.01). In the normal groups, statistical analysis revealed that there were significant differences in the percentage of S + G2M phase, a value of MTT and the expression rate of PCNA protein in three groups (P <0.01). Sense CyclinD1 eukaryotic expression vectors could have a positive effect on the proliferation of ASMC, however the antisence one have a negative effect, which implicated that CyclinD1 might contribute to the process of airway smooth muscle cell proliferation.
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Asma/patologia , Proliferação de Células/efeitos dos fármacos , Ciclina D1/antagonistas & inibidores , DNA Antissenso/farmacologia , Miócitos de Músculo Liso/efeitos dos fármacos , Animais , Ciclo Celular/efeitos dos fármacos , Códon/genética , Códon/farmacologia , Ciclina D1/agonistas , Ciclina D1/genética , DNA Antissenso/genética , Modelos Animais de Doenças , Expressão Gênica , Vetores Genéticos/genética , Masculino , Miócitos de Músculo Liso/patologia , Ratos , Ratos Sprague-Dawley , Recombinação Genética/genética , Sistema Respiratório , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Transdução Genética , TransfecçãoRESUMO
OBJECTIVE: To explore the role of PKCalpha-ERK1/2 cascade in PMA induced up-regulation of cyclinD1 and P21(cip1) in human airway smooth muscle cells (HASMCs) sensitized by sera from atopic asthmatics. METHODS: HASMCs in cultures were passively sensitized by 10% serum from asthmatic patients and were randomly divided into five groups: the control group, PMA treated group, PMA and PKCalpha mismatched Oligodeoxynucleotides (PKCalpha-mmODN) treated group, PMA and PKCalpha antisense Oligodeoxynucleotides (PKCalpha-asODN) treated group, PMA and U0126 (MAP Kinase Kinase inhibitor)treated group. The expression of p-PKCalpha, ERK1/2, p-ERK1/2, cyclinD1 and P21(cip1) protein were determined by western blotting. The proliferation of HASMC was examined by cell cycle analysis and MTT colorimetric assay. RESULTS: Compared with the control group, the expression of p-PKCalpha and ERK1/2, p-ERK1/2 protein increased, the expression of cyclinD1, P21(cip1) protein increased correspondingly (the A value % control was 2.10 +/- 0.29, 1.67 +/- 0.19, 2.20 +/- 0.27, 1.99 +/- 0.22 and 3.11 +/- 0.29 respectively; q value was 9.87, 7.06, 10.57, 11.10 and 20.33 respectively; all P < 0.05) in PMA treated group, and cells proliferation [the percentage of cells in S phase was (30.3 +/- 2.4)%, A(490) value was 0.80 +/- 0.06] enhanced significantly compared with those [the percentage of cells in S phase was (13.9 +/- 2.6)%, A(490) value was 0.41 +/- 0.04] of the control group (q = 6.07, 12.63; all P < 0.05). In PMA and PKCalpha-asODN treated group, the level of p-PKCalpha decreased, the expression of ERK1/2, p-ERK1/2 and the expression of cyclinD1, P21(cip1) decreased correspondingly (the A value % control was 1.23 +/- 0.19, 1.34 +/- 0.18, 1.52 +/- 0.20, 1.45 +/- 0.18 and 1.49 +/- 0.18 respectively; q value was 7.49, 3.58, 5.97, 6.06 and 15.65 respectively; all P < 0.05), and cells proliferation reduced significantly [the percentage of cells in S phase was (21.2 +/- 2.8)%, A(490) value was 0.51 +/- 0.04; q = 6.07, 12.63; all P < 0.05], as compared with those of the PMA treated group. In PMA and U0126 treated group, the level of p-PKCalpha had no significant change (A value was1.99 +/- 0.18, q = 0.94, P > 0.05), but the levels of ERK1/2, p-ERK1/2 decreased, the expression of cyclinD1, P21(cip1) reduced (the A value % control was 0.95 +/- 0.21, 1.15 +/- 0.19, 1.37 +/- 0.15 and 1.96 +/- 0.21 respectively; q value was 7.79, 9.16, 6.92 and 11.16 respectively; all P < 0.05), and cells proliferation reduced significantly [the percentage of cells in S phase was (22.0 +/- 3.2)%, A(490) value was 0.49 +/- 0.03; q = 5.51, 13.45; all P < 0.05], as compared with those of the PMA treated group. CONCLUSION: ERK1/2 is one of the downstream regulators of PKCalpha, and PKCalpha-ERK1/2 cascade is involved in PMA induced up-regulation of cyclinD1 and P21(cip1) and proliferation in HASMC sensitized by sera from atopic asthmatics.
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Asma/metabolismo , Ciclina D1/metabolismo , Inibidor de Quinase Dependente de Ciclina p21/metabolismo , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Miócitos de Músculo Liso/metabolismo , Proteína Quinase C-alfa/metabolismo , Adulto , Asma/sangue , Ciclo Celular , Proliferação de Células , Células Cultivadas , Feminino , Regulação da Expressão Gênica , Humanos , Masculino , Pessoa de Meia-Idade , Proteína Quinase 3 Ativada por Mitógeno/metabolismo , Miócitos de Músculo Liso/citologia , Transdução de Sinais , Regulação para Cima , Adulto JovemRESUMO
Following publication of their article, the authors noticed that there were minor errors in Figs. 3, 7 and S5. The errors had no effect on the scientific content or conclusions. The rectified figures are given below.
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This corrects the article DOI: 10.1038/cddis.2016.260.
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Vacinas contra COVID-19 , COVID-19/imunologia , Imunidade Humoral/efeitos dos fármacos , Rinite Alérgica/imunologia , SARS-CoV-2/imunologia , Vacinas de Produtos Inativados , Adulto , COVID-19/prevenção & controle , Vacinas contra COVID-19/administração & dosagem , Vacinas contra COVID-19/imunologia , Feminino , Humanos , Masculino , Estudos Prospectivos , Vacinas de Produtos Inativados/administração & dosagem , Vacinas de Produtos Inativados/imunologiaRESUMO
The number of smokers in Chinese rural areas is more than 200 million, which is twice that in cities. It is very significant to carry out tobacco control interventions in rural areas. We performed this community intervention study to evaluate the efficacy of village-based health education of tobacco control on the male current smoking rate in rural areas. The population of this study was the males above 15 years old from 6 villages in rural areas. The villages were randomly assigned to intervention group or control group (3 villages in each group). Self-designed smoking questionnaire was applied. The intervention group received the village-based health education of tobacco control for one year. The primary outcome measurement was the male current smoking rate. In the baseline investigation, completed surveys were returned by 814 male residents from the control group and 831 male residents from the intervention group. The male current smoking rate in the control group and the intervention group was 61.2% and 58.5%, respectively, before intervention. There was no significant difference between these two groups (P>0.05). After one-year intervention, the current smoking rate in the intervention group (51.2%) was significantly lower than that in the control group (62.8%) (P<0.001). Our study suggested that the village-based health education of tobacco control was effective in lowering the male current smoking rate in rural areas, which could be a suitable and feasible way for tobacco control in the Chinese rural areas.
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Educação em Saúde/métodos , População Rural , Prevenção do Hábito de Fumar , Abandono do Uso de Tabaco , Adolescente , Adulto , Estudos de Casos e Controles , China , Atenção à Saúde/métodos , Humanos , Masculino , Pessoa de Meia-IdadeRESUMO
Our previous study revealed that Ku80 was overexpressed in lung cancer tissues and hsa-miR-623 regulated the Ku80 expression; however, the detailed function of hsa-miR-623 in lung cancer was unclear. We identified that hsa-miR-623 bound to the 3'-UTR of Ku80 mRNA, thus significantly decreasing Ku80 expression in lung adenocarcinoma cells. Hsa-miR-623 was downregulated in lung adenocarcinoma tissues compared with corresponding non-tumorous tissues, and its expression was inversely correlated with Ku80 upregulation. Downregulation of hsa-miR-623 was associated with poor clinical outcomes of lung adenocarcinoma patients. Hsa-miR-623 suppressed lung adenocarcinoma cell proliferation, clonogenicity, migration and invasion in vitro. Hsa-miR-623 inhibited xenografts growth and metastasis of lung adenocarcinoma in vivo. Ku80 knockdown in lung adenocarcinoma cells suppressed tumor properties in vitro and in vivo similar to hsa-miR-623 overexpression. Further, hsa-miR-623 overexpression decreased matrix metalloproteinase-2 (MMP-2) and MMP-9 expression levels, with decreased ERK/JNK phosphorylation. Inhibition of hsa-miR-623 or overexpression of Ku80 promoted lung adenocarcinoma cell invasion, activated ERK/JNK phosphorylation and increased MMP-2/9 expressions, which could be reversed by ERK kinase inhibitor or JNK kinase inhibitor. In summary, our results showed that hsa-miR-623 was downregulated in lung adenocarcinoma and suppressed the invasion and metastasis targeting Ku80 through ERK/JNK inactivation mediated downregulation of MMP-2/9. These findings reveal that hsa-miR-623 may serve as an important therapeutic target in lung cancer therapy.
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Adenocarcinoma/genética , Adenocarcinoma/patologia , Progressão da Doença , Neoplasias Pulmonares/genética , Neoplasias Pulmonares/patologia , MicroRNAs/metabolismo , Adenocarcinoma de Pulmão , Animais , Sequência de Bases , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Regulação para Baixo/genética , Feminino , Regulação Neoplásica da Expressão Gênica , Humanos , Autoantígeno Ku/metabolismo , Sistema de Sinalização das MAP Quinases , Masculino , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 9 da Matriz/metabolismo , Camundongos Endogâmicos BALB C , Camundongos Nus , MicroRNAs/genética , Pessoa de Meia-Idade , Invasividade Neoplásica , Metástase Neoplásica , Resultado do Tratamento , Ensaios Antitumorais Modelo de XenoenxertoRESUMO
OBJECTIVE: To investigate whether polymorphism in gene for surfactant protein A(SPA) has any bearing on individual susceptibility to the development of chronic obstructive pulmonary disease(COPD). METHODS: The genotypes of 88 patients with COPD and 87 healthy smoking subjects as controls were tested with polymerase chain reaction followed by restriction fragment length polymorphism analysis for SPA gene. RESULTS: In COPD group, the frequencies of +186 locus genotypes AA, AG and GG were 86.4%, 12.5% and 1.1%i respectively and in the control group, these were 66.7%, 27.6% and 5.7%; the frequencies of polymorphic genotypes or alleles showed statistically significant difference between the COPD group and the control group(P<0.05). The frequencies of polymorphic genotypes at +655 locus and +667 loci showed no significant difference between the COPD group and the control group in(P>0.05). CONCLUSION: Genetic polymorphism in SPA is associated with the development of COPD in the Chinese Hans.
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Polimorfismo Genético , Doença Pulmonar Obstrutiva Crônica/genética , Proteína A Associada a Surfactante Pulmonar/genética , Idoso , Idoso de 80 Anos ou mais , Alelos , Feminino , Frequência do Gene , Predisposição Genética para Doença/genética , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da PolimeraseRESUMO
OBJECTIVE: To explore the relationship of DNA damage in lymphocytes with chronic obstructive pulmonary disease (COPD). METHODS: Levels of heat shock protein 70 (HSP70) in peripheral lymphocytes were measured by Western blot in 20 COPD patients with smoking, 20 healthy smoking control subjects and 15 healthy nonsmokers. DNA damage in the lymphocytes was detected by Comet Assay. RESULTS: DNA damage score was 42.1 +/- 10.2, 17.4 +/- 9.3 and 10.1 +/- 3.6 in COPD patients, healthy smoking control subjects and healthy nonsmokers, respectively; the score was higher in COPD patients than that in healthy smoking control subjects, and was higher in smokers than in nonsmokers (P < 0.01). The levels of HSP70 (absorbency value) were 20.9 +/- 9.9, 44.8 +/- 15.3 and 56.1 +/- 18.9, respectively, and the difference was significant among the three groups (P < 0.05). The level of HSP70 showed a strongly negative correlation with DNA damage scores (r = -0.73, P < 0.01). CONCLUSION: DNA damage may be involved in the pathogenesis of COPD. The difference of HSP70 level may be associated with the level of DNA damage.
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Dano ao DNA , Doença Pulmonar Obstrutiva Crônica/genética , Idoso , Western Blotting , Ensaio Cometa , Feminino , Proteínas de Choque Térmico HSP70/metabolismo , Humanos , Linfócitos/metabolismo , Linfócitos/patologia , Masculino , Pessoa de Meia-Idade , Doença Pulmonar Obstrutiva Crônica/metabolismo , FumarRESUMO
OBJECTIVE: To determine whether 2 polymorphism sites of the high-affinity IgE receptor beta (Fc epsilon RI beta) gene were associated with allergic asthma in Han nationality of Hubei province in China. METHODS: DNA and clinical data were obtained from allergic asthma patients and were compared with those from a group of healthy control subjects. The subjects were genotyped for the -109C/T and a coding variant Glu237Gly in Fc epsilon RI beta gene by polymerase chain reaction-restriction fragment length polymorphism. RESULTS: (1) The genotype frequencies were 0.403 for -109T/T, 0.491 for -109T/C and 0.106 for -109C/C in allergic asthma patients in a Chinese population. No significant difference in the distribution of -109C/T polymorphism was found between allergic asthma patients and healthy control subjects; however, a homozygosity for the -109T allele was associated with increased total plasma IgE levels in patients with allergic asthma (F=7.523, P<0.01). (2) The frequency of Gly237Gly genotype was 0.051 in patient group, and 0.020 in control group. The allele frequencies of Gly237 in the patients and control were 0.236 and 0.136 respectively. There was a significant association between Gly/Gly genotype and allergic asthma. Among allergic asthma patients Gly237Gly was significantly associated with high IgE. CONCLUSION: These data suggested that the Gly237Gly genotype of the Fc epsilon RI beta gene conferred genetic susceptibility to allergic asthma in Chinese, which affected the total plasma IgE levels in the allergic asthma patients. And a homozygosity for the -109T allele was associated with increased total plasma IgE.
Assuntos
Asma/genética , Polimorfismo Genético , Receptores de IgE/genética , Adolescente , Adulto , Idoso , Povo Asiático/genética , Asma/sangue , Criança , Pré-Escolar , China , Feminino , Frequência do Gene , Predisposição Genética para Doença/genética , Genótipo , Humanos , Imunoglobulina E/sangue , Masculino , Pessoa de Meia-Idade , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Regiões Promotoras Genéticas/genética , Adulto JovemRESUMO
OBJECTIVE: To investigate the polymorphism of interleukin-10 (IL-10) gene promoters in Chinese Han people, and to disclose whether such polymorphism is associated with susceptibility to chronic obstructive pulmonary disease (COPD). METHODS: After the process of extracting genomic DNA from blood of 94 health smokers and 88 COPD smokers by use of phenol-chloroform-isoamyl alcohol, three single nucleotide polymorphism (SNP) sites in IL-10 gene promoter marked as -1082G/A,-819C/T,-592C/A were determined by polymerase chain reaction/restriction fragment length polymorphism analysis. RESULTS: Eleven different promoter genotypes were detected from all of the 182 smokers, and AA*TT*AA, AA*TC*AC, AA*TC*AA genotypes accounted for about 80% of genotypes in the research subjects. Two previously unreported haplotypes of IL-10 gene promoter (ATC and ACA) were found in Chinese Han people by analyzing the promoter genotypes. -1082G/A and -592C/A SNP sites polymorphisms were not associated with susceptibility to COPD, whereas the genotypes of -819C/T SNP site were associated with susceptibility to COPD in Chinese Han people. In respect to the alleles frequencies of the three SNP sites respectively, the Chinese Hans were similar to Japanese, but different from whites. CONCLUSION: Polymorphism of IL-10 -819C/T SNP site is associated with susceptibility to COPD in Chinese Han people; at least five haplotypes of IL-10 gene promoter (ATA, ACC, GCC, ATC and ACA) exist in Chinese Han people.