The Crosstalk between lncRNA-SNHG7/miRNA-181/cbx7 Modulates Malignant Character in Lung Adenocarcinoma.

Lung adenocarcinoma (LUAD) is a malignant tumor with poor patient survival and high patient mortality. Long noncoding RNA is profoundly involved in the tumorigenesis of LUAD. The present study explores the effect of small nucleolar RNA host gene 7 (SNHG7) on the progression of LUAD and its underlying mechanisms. SNHG7 was found to be down-regulated in LUAD tissues compared with normal tissues. Altered SNHG7 expression induced changes in cell proliferation and migration both in vitro and in vivo. Mechanistically, it was found that SNHG7 interacted with microRNA mir-181 and sequentially up-regulated cbx7. cbx7, which suppresses the Wnt/ß-catenin pathway in LUAD, was found to be a direct target of mir-181. Taken together, loss of SNHG7 in LUAD up-regulated mir-181 and then down-regulated the tumor suppressor cbx7.

Aberrant promoter methylation of cell adhesion-related genes associated with clinicopathologic features in non-small cell lung cancer in China.

PURPOSE: The aim of this study was to investigate the methylation status of three cell adhesion-related genes including CDH1, TSLC1 and TIMP3 in non-small cell lung cancer and explore its association with clinicopathologic features and various environmental risk factors. METHODS: We detected the aberrant methylation presence of these genes by methylation-specific polymerase chain reaction and analyzed the potential correlations with multivariate logistic regression model as well as stepwise logistic regression. RESULTS: For CDH1, promoter methylation was less frequent in adenosquamous carcinomas than adenocarcinomas (OR=0.35, 95%CI=0.13-0.96); pickled food increased the methylation frequency (OR=2.23, 95%CI=1.09-4.54) while light smoking and fruit intake decreased that (OR=0.43, 95%CI=0.19-0.97; OR=0.37, 95%CI=0.15-0.95). For TSLC1, males and toxin exposure increased methylation frequency (OR=6.25, 95%CI=1.05-37.13; OR=2.42, 95%CI=1.01-5.77) while light smoking and radiation exposure decreased that (OR=0.14, 95%CI=0.03-0.60; OR=0.17, 95%CI=0.04-0.87). For TIMP3, males showed lower methylation frequency than females (OR=0.18, 95%CI=0.04-0.88) while central lung cancer, heavy smoking and radiation exposure presented higher aberrant DNA methylation status (OR=2.19, 95%CI=1.07-4.52; OR=6.99, 95%CI=1.32-37.14; OR=2.30, 95%CI=1.04-5.08). CONCLUSIONS: Aberrant promoter methylation of cell adhesion-related tumor suppressor genes in lung cancer displayed varieties of gene-specific correlations with clinicopathologic features and various environmental risk factors.

[Correlation between RARbeta gene promoter methylation and P53 gene mutations in non-small cell lung cancer].

OBJECTIVE: To investigate the correlation between RARbeta gene promoter methylation and P53 gene mutations in non-small cell lung cancer (NSCLC). METHODS: Promoter methylation of RARbeta and P53 mutations of exons 5 through 9 in 198 resected primary NSCLC tissues were determined by methylation-specific PCR and direct sequencing. RESULTS: RARbeta gene promoter methylation and P53 mutation were detected in 58.1% and 36.4% of tumors, respectively. Both were higher in males than in females and in smokers than in nonsmokers. A higher prevalence of RARbeta promoter methylation was found in patients with advanced stage tumors than those with TNM stage I. P53 gene mutations were more frequent in squamous cell carcinoma and adeno-squamous carcinoma than adenocarcinoma. All such differences were statistically significant (P< 0.05). Frequencies of P53 mutations, including G:C>T:A mutations, transversions and missense mutations were significantly higher in tumors with RARbeta methylation than in those without (P< 0.05). A significantly higher prevalence of RARbeta methylation was found in tumors with only G:C>T:A mutation in P53 gene than those without P53 mutations (P< 0.05). This difference (OR=3.737, 95%CI: 1.414-9.873) was still statistically significant (P< 0.05) in smokers (OR=4.020, 95%CI: 1.263-12.800), squamous cell carcinomas (OR=5.480, 95%CI: 1.400-21.446) or patients with advanced tumors (OR=3.446, 95%CI: 1.054-11.267) after adjusting for age and sex. CONCLUSION: RARbeta methylation is associated with G:C>T:A mutations in P53 gene in NSCLC.

[Effects of CYP1A1 and GSTM1 gene polymorphisms and BPDE-DNA adducts on lung cancer].

OBJECTIVE: To investigate the effect of CYP1A1 and GSTM1 genetic polymorphisms and BPDE-DNA adducts on lung tumorigenesis. METHODS: The case control study has included 200 cases of lung cancer and 200 controls. DNA was extracted from blood samples of all subjects. The genotype of both CYP1A1 and GSTM1 were detected with PCR-based restriction fragment length polymorphisms (PCR-RELP). BPDE-DNA adducts were detected with competitive ELISA. RESULTS: CYP1A1 mutant genotype and GSTM1 null genotype with smoke has increased the risk of lung cancer, with OR being 2.406(1.321-4.382), 2.755(1.470-5.163), respectively. The level of BPDE-DNA adducts in patients was greater than control, and the adduct level in ever smokers was higher than never smokers, the difference was statistically significant (P= 0.0252). GSTM1 null genotype individuals with BPDE-DNA level higher than 5 adducts/10(8) nucleotide have increased risk of lung cancer (OR= 1.988, 95%CI: 1.011-3.912). Compared with never smokers with CYP1A1 wild genotype, smokers with CYP1A1 mutation genotype had an increased risk of forming a higher level of DNA adducts (P= 0.0459). Smokers with GSTM1 null genotype formed more DNA adducts compared with never smokers with GSTM1 functional genotype (OR = 2.432, 95% CI: 1.072-4.517). CONCLUSION: GSTM1 null genotype with higher level DNA adducts may increase the risk of lung cancer. DNA adducts form easier in smokers with CYP1A1 mutation genotype and GSTM1 null genotype, which in turn may influence lung tumorigenesis.

Effect of CYP1A1 MSPI polymorphism on the relationship between TP53 mutation and CDKN2A hypermethylation in non-small cell lung cancer.

BACKGROUND AND AIMS: The molecular mechanisms of lung cancer susceptibility have not been fully understood. Although it has been described that germline polymorphisms are associated with either mutation or methylation of genes, the link between gene polymorphisms and gene-gene interactions has not been investigated. Therefore, we conducted this study to determine whether CYP1A1/GSTM1 polymorphisms can affect the relationship between TP53 mutation and CDKN2A hypermethylation in lung cancer. METHODS: This study included 196 primary non-small cell lung cancer (NSCLC) patients. CYP1A1 MSPI and GSTM1 polymorphisms were characterized through PCR-RFLP on DNA isolated from peripheral lymphocytes. TP53 mutations of exons 5 through 9 and CDKN2A promoter hypermethylation in both cancer tissues and corresponding normal tissues were analyzed by direct sequencing and methylation-specific PCR (MSP) respectively. RESULTS: TP53 mutation in the tumor was associated with squamous cell histology and CDKN2A methylation was associated with older age (≥60 years), heavy smoking (>30 pack-years), squamous cell histology and advanced stage (stage II-IV). After adjusting for age, sex, smoking degree, histology type and TNM stage, the correlation between TP53 mutation and CDKN2A methylation was significant in patients with CYP1A1 risk genotype (p = 0.038), but not in those with CYP1A1 homogeneity wild genotype (p = 0.151). CONCLUSIONS: This may suggest that TP53 mutation and CDKN2A methylation specifically interact to promote lung tumorigenesis in subjects with CYP1A1 risk genotype but not in those with CYP1A1 wild-type homozygotes, implying different pathways for the development of lung carcinoma with respect to CYP1A1 polymorphism.

Addition of ulinastatin to preservation solution promotes protection against ischemia-reperfusion injury in rabbit lung.

BACKGROUND: The composition of the lung preservation solution used in lung graft procurement has been considered the key to minimize lung injury during the period of ischemia. Low-potassium dextran glucose (LPDG), an extracellular-type solution, has been adopted by most lung transplantation centers, due to the experimental and clinical evidences that LPDG is superior to intracellular-type solutions. Ulinastatin has been shown to attenuate ischemia-reperfusion (I/R) injury in various organs in animals. We supposed that the addition of ulinastatin to LPDG as a flushing solution, would further ameliorate I/R lung injury than LPDG solution alone. METHODS: Twelve male New Zealand white rabbits were randomly divided into 2 groups. Using an alternative in situ lung I/R model, the left lung in the control group was supplied and preserved with LPDG solution for 120 minutes. In the study group 50,000 U/kg of ulinastatin was added to the LPDG solution for lung preservation. Then re-ventilation and reperfusion of the left lung were performed for 90 minutes. Blood gas analysis (PaO2, PaCO2), mean pulmonary artery pressure (MPAP) and serum TNF-α level were measured intermittently. The pulmonary water index (D/W), tissue myeloperoxidase (MPO) activity, tissue malondialdehyde (MDA) content and morphologic changes were analyzed. RESULTS: The study group showed significantly higher PaO2 and lower MPAP at the end of reperfusion. Serum TNF-α level, left lung tissue MPO and MDA in the study group were significantly lower than those in the control group. D/W and pathologic evaluation were also remarkably different between the two groups. CONCLUSIONS: This study indicated that better lung preservation could be achieved with the use of an ulinastatin modified LPDG solution. Ulinastatin further attenuated lung I/R injury, at least partly by reducing oxidative reactions, inhibiting the release of inflammatory factors and neutrophils immigration.

[Relationship between promoter methylation of p16, DAPK and RAR beta genes and the clinical data of non-small cell lung cancer].

OBJECTIVE: To investigate the effects of promoter methylation of p16, death-associated protein kinase (DAPK) and retinoic acid receptor-beta (RAR beta) genes on clinical data in non-small cell lung cancers, and to study the effect of smoking on the risk of gene methylation. METHODS: The promoter methylation of p16, DAPK and RAR beta genes in 200 primary non-small cell lung cancers and the corresponding nonmalignant lung tissues were determined by methylation-specific PCR. RESULTS: Methylation in the tumor tissues was detected in 51.0% for p16, 60.0% for DAPK, and 58.0% for RAR beta gene, with significant differences (P < 0.05) when compared with those in the corresponding nonmalignant tissues(12.5%, 11.5% and 15.0%) respectively. p16 gene methylation in tumor tissue was associated with age significantly in unconditional logistic regression analysis (P < 0.01) and histologic type (P < 0.05). DAPK gene methylation in tumor tissue was associated significantly with age (P < 0.05), gender (P < 0.05) and clinical type (P < 0.05). RAR beta gene methylation in tumor tissue was associated with clinical type (P < 0.05) and tumor stage (P < 0.05) significantly. The interaction odds ratio (OR) for the gene-gene interaction in tumor tissue between p16 and DAPK was 1.987 (95%CI:1.055-3.743). The results of the gene-smoking analyses revealed that a relationship existed between cigarette smoking and p16 gene methylation (OR = 3.139, 95%CI: 1.046-9.419), the OR for the relationship of DAPK gene methylation and cigarette smoking was 3.585(95%CI: 1.270-10.123) in tumor tissue. The RAR beta gene methylation did not differ based on the smoking status of patients in tumor tissue. CONCLUSION: The p16, DAPK and RAR beta genes methylation are strongly associated with clinical data of non-small cell lung cancer, and methylation of p16 and DAPK genes are associated with tobacco smoking.

Diagnosis and treatment of pulmonary mucosa-associated lymphoid tissue lymphoma.

BACKGROUND: Primary non-Hodgkin's lymphoma in lung is very rare, and the most common among them is mucosa-associated lymphoid tissue lymphoma (MALToma), whose clinical features and laboratory characteristics are poorly defined, making diagnosis difficult. The purpose of this study was to study the diagnosis and treatment of pulmonary MALToma. METHODS: The clinical data of 12 patients treated for MALToma between August 1992 and December 2005 were analyzed. RESULTS: No specific symptoms or signs, or results of bronchoscopy, ultrasonagraphy or bone marrow examination could be found in the 12 patients. Only radiography was useful in diagnosis, though the final diagnosis of all the patients was based on histology and immunohistochemistry. Two patients also had gastric MALToma. Operations were performed on 6 patients, including 5 radical operations and 1 partial resection: 4 patients also received adjuvant chemotherapy. One patient experienced recurrence 152 months after the operation, while the other 5 patients have survived disease-free. Four patients were treated with chemotherapy alone, two of whom experienced complete remission and the others partial remission. The final 2 patients received no treatment and had survived for 7 and 27 months respectively. All the patients were still alive at the most recent follow-up, 7 to 160 months (mean 71.3 months). CONCLUSIONS: Except radiography, no specific clinical manifestations could be identified for pulmonary MALToma. The final diagnosis should be based on histology and immunohistochemistry. Several treatment methods can be used to achieve good outcomes.

[Selective determination of dopamine in the presence of high concentration of ascorbic acid with L-cysteine modified glassy carbon electrodes].

OBJECTIVE: To establish an assay system for determination of dopamine (DA) in the presence of ascorbic acid (AA) with L-cysteine modified glassy carbon electrode. METHODS: L-cysteine was modified onto glassy carbon electrode electrochemically, and with this modified electrode, dopamine was determined by linear sweep stripping voltammetry. RESULTS: L-cysteine polymer-modified electrode had strong catalytic effect towards the electrochemical oxidation of DA. The modified electrode showed good properties in determination of DA with coexisting AA. Under selected conditions, the linearity of DA was in the range of 2.0 x 10(-7) - 1.0 x 10(-4) mol/L with the detection limit of 2.0 x 10(-8) mol/L. The stability, reliability and recovery of this L-cysteine-modified electrode based on electrochemical method were also satisfactory. CONCLUSION: L-cysteine-modified electrode can avoid the interference by AA for determination of DA.

[Protective effects of ginaton against ischemia-reperfusion injury on the autograft after lung autotransplantation: experiment with rabbits].

OBJECTIVE: To investigate the effects of ginaton against ischemia-reperfusion injury on the autograft after lung autotransplantation. METHODS: Models of lung autotransplantation were established in 18 New Zealand rabbits were established. The 18 rabbits were randomly divided into 3 equal groups: group of simple ischemia-reperfusion (group I/R), undergoing ischemia by blocking the left pulmonary artery for 2 h and then re-perfusion for 90 min; group with perfusion of low potassium dextran solution (group LPD), undergoing perfusion of LSD solution before ischemia; and group with treatment of ginaton (group LPD + E), undergoing intravenous injection of ginaton 15 min before ischemia. Arterial blood samples were collected before ischemia, and 15, 60, and 90 min after re-perfusion to examine the alveolar oxygen pressure (PaO2). Serum tumor necrosis factor-alpha (TNF-alpha) was monitored before ischemia, and 30, 60, and 90 min after re-perfusion. Then the left lungs were taken out to undergo detection of dry/wet ratio (D/W), pathological examination, and contents of myeloperoxidase (MPO) and the malondialdehyde (MDA) in the lung tissues. RESULTS: (1) The PaO2 decreased significantly after reperfusion in all groups. And the PaO2 values at different time points of Group LPD + E were all significantly higher than those of Group I/R (all P < 0.01), however, there were no significant differences between Group LPD and Group LPD + E. (2) The TNF-alpha level after reperfusion increased in Group I/R and Group LPD, while in Group LPD + E it increased only 60 min and 90 min after the reperfusion. The TNF-alpha levels after reperfusion at all time points of Group LPD + E were all significantly lower than those of the other 2 groups (all P < 0.05). (3) The MPO and MDA levels at all time points after re-perfusion of Group LPD + E were all significantly lower than those of the other 2 groups (all P < 0.01). (4) The value of D/W ratio of Group LPD + E was significantly higher than those of the other 2 groups (both P < 0.01). (5) Pathological examination showed that the lung tissue lesion of Group I/R was severe. Interstitial inflammatory cell infiltration, intra-alveolar inflammatory cell aggregation, exudation and even hemorrhage could be observed. The pathological lesion of Group LPD + E was mild, no significant inflammatory cell infiltration or exudation was observed. CONCLUSION: Ginaton provides a protective effect against ischemia-reperfusion injury on the autograft after lung autotransplantation. The mechanism may be related with antioxidation, inhibition of neutrophil aggregation, and TNF-releasing.

[Diagnosis and treatment of primary pulmonary mucosa-associated lymphoid tissue lymphoma].

OBJECTIVE: To study the diagnosis and treatment for primary pulmonary mucosa-associated lymphoid tissue lymphoma. METHODS: The clinical data of 12 patients with primary pulmonary mucosa-associated lymphoid tissue lymphoma from August 1992 to May 2005 were analyzed. RESULTS: All the patients survived during the follow-up periods of 6 to 164 months (mean 70.3 months). Gastric mucosa-associated lymphoid tissue lymphoma was found to coexist in 2 patients. No specific symptoms or signs, or specific results of bronchoscopy, ultrasonography or bone marrow examination were found in these patients, except that radiography showed nodules with blurred margins with characteristic air bronchogram. The final diagnosis was based on histology and immunohistochemistry. Surgical resection was performed for 6 patients, including 5 radical operations and 1 partial resection, among which 4 patients received adjuvant chemotherapy. Recurrence occurred in 1 patient 12.7 years after the operation, while the other 5 patients got disease free survival. Chemotherapy alone was administered for 4 patients, among whom 2 patients got complete remission and the others got partial remission. The other 2 patients received no treatment and had survived for 6 and 26 months respectively. CONCLUSIONS: Except for the radiographic findings, there were no specific clinical manifestations for primary pulmonary mucosa-associated lymphoid tissue lymphoma. The final diagnosis should be made by histology and immunohistochemistry. Surgery and chemotherapy can be adopted for the patients with good outcomes.