Objective:Recent research has indicated that altered promoter methylation of oncogenes and tumor suppressor genes is an important mechanism in lung cancer development and progression.In this study,we investigated the ...Objective:Recent research has indicated that altered promoter methylation of oncogenes and tumor suppressor genes is an important mechanism in lung cancer development and progression.In this study,we investigated the association between promoter methylation of TMEM88,a possible inhibitor of the Wnt/β-Catenin signaling,and the survival of patients with nonsmall cell lung cancer(NSCLC).Methods:Twelve pairs of tumor and adjacent non-tumor samples were used for microarray analyses of DNA methylation and gene expression.For validation,more than two hundred additional samples were analyzed for methylation using bisulfite pyrosequencing and for gene expression using q RT-PCR.Then the cell function were tested by wound healing,transwell,CCK8 and cell cycle assay.Results:Our analysis of patient specimens showed that TMEM88 methylation was higher in NSCLC tumors(82.2%±10.3,P<0.01)compared with the adjacent normal tissues(65.9%±7.2).The survival analysis revealed that patients with high TMEM88methylation had a shorter overall survival(46 months)compared with patients with low TMEM88 methylation(>56 months;P=0.021).In addition,we found that demethylation treatment could inhibit tumor cell proliferation,migration,and invasion,which was supportive of an association between methylation and survival.Conclusions:Based on these consistent observations,we concluded that TMEM88 may play an important role in NSCLC progression and that promoter methylation of TMEM88 may serve as a biomarker for NSCLC prognosis and treatment.展开更多
目的探讨跨膜蛋白(transmembrane protein 88,TMEM88)对缺血-再灌注小鼠心肌纤维化(myocardial fibrosis,MF)的影响及可能机制研究。方法将36只SPF级C57小鼠随机分为对照组(Control组)、心肌缺血-再灌注模型组(Ischemia reperfusion,I/R...目的探讨跨膜蛋白(transmembrane protein 88,TMEM88)对缺血-再灌注小鼠心肌纤维化(myocardial fibrosis,MF)的影响及可能机制研究。方法将36只SPF级C57小鼠随机分为对照组(Control组)、心肌缺血-再灌注模型组(Ischemia reperfusion,I/R组)和心肌缺血-再灌注模型+TMEM88组(I/R+TMEM88组)。采用对左冠状动脉前降支结扎的方法构建缺血-再灌注小鼠模型。利用超声心动图检测各组小鼠心功能;Masson染色检测各组小鼠心脏病理学变化并测定胶原容积分数(Collagen volume fraction,CVF);蛋白免疫印迹(Western blot)法检测β-连环蛋白(β-catenin)、α-平滑肌肌动蛋白(alpha-smooth muscle actin,α-SMA)和E-钙黏蛋白(E-cadherin)的表达情况。结果与Control组比较,I/R组小鼠LVEDD和LVEDV明显增高[LVEDD(mm):3.37±0.25 vs. 4.28±0.56,LVEDV(mm):46.89±8.20 vs. 83.80±22.48],LVFS和LVEF明显降低[(LVFS:(%) 30.33±3.18 vs. 20.71±2.82,LVEF(%):58.77±4.55vs. 42.44±4.80],CVF升高(%:4.06±0.74 vs. 29.54±1.32),β-catenin及α-SMA表达明显增加(β-catenin:0.35±0.17 vs. 0.74±0.14,α-SMA:0.39±0.21 vs. 0.85±0.30),Ecadherin表达减少(0.54±0.16 vs. 0.28±0.10),差异均有统计学意义(P<0.05);与I/R组比较,I/R+TMEM88组小鼠LVEDD和LVEDV减低[LVEDD(mm):4.28±0.56 vs. 3.85±0.29,LVEDV(mm):83.80±22.48 vs. 64.27±11.59],LVFS和LVEF增高[LVFS(%):20.71±2.82vs. 24.14±3.79,LVEF(%):42.44±4.80 vs. 48.53±6.14),CVF降低(%:29.54±1.32 vs.25.06±3.62),β-catenin及α-SMA表达减少(β-catenin:0.74±0.14 vs. 0.58±0.10,α-SMA:0.85±0.30 vs. 0.59±0.11),E-cadherin表达增加(0.28±0.10 vs. 0.41±0.05),差异均有统计学意义(P<0.05)。结论 TMEM88能够通过抑制Wnt/β-catenin信号通路活性改善缺血-再灌注小鼠的心肌纤维化程度。展开更多
基金supported by the National Natural Science Foundation of China(Grant No.81573231)the Medical Professionals Crossing Project of Shanghai Jiao Tong University(Grant No.YG2015ZD01)
文摘Objective:Recent research has indicated that altered promoter methylation of oncogenes and tumor suppressor genes is an important mechanism in lung cancer development and progression.In this study,we investigated the association between promoter methylation of TMEM88,a possible inhibitor of the Wnt/β-Catenin signaling,and the survival of patients with nonsmall cell lung cancer(NSCLC).Methods:Twelve pairs of tumor and adjacent non-tumor samples were used for microarray analyses of DNA methylation and gene expression.For validation,more than two hundred additional samples were analyzed for methylation using bisulfite pyrosequencing and for gene expression using q RT-PCR.Then the cell function were tested by wound healing,transwell,CCK8 and cell cycle assay.Results:Our analysis of patient specimens showed that TMEM88 methylation was higher in NSCLC tumors(82.2%±10.3,P<0.01)compared with the adjacent normal tissues(65.9%±7.2).The survival analysis revealed that patients with high TMEM88methylation had a shorter overall survival(46 months)compared with patients with low TMEM88 methylation(>56 months;P=0.021).In addition,we found that demethylation treatment could inhibit tumor cell proliferation,migration,and invasion,which was supportive of an association between methylation and survival.Conclusions:Based on these consistent observations,we concluded that TMEM88 may play an important role in NSCLC progression and that promoter methylation of TMEM88 may serve as a biomarker for NSCLC prognosis and treatment.
文摘目的探讨跨膜蛋白(transmembrane protein 88,TMEM88)对缺血-再灌注小鼠心肌纤维化(myocardial fibrosis,MF)的影响及可能机制研究。方法将36只SPF级C57小鼠随机分为对照组(Control组)、心肌缺血-再灌注模型组(Ischemia reperfusion,I/R组)和心肌缺血-再灌注模型+TMEM88组(I/R+TMEM88组)。采用对左冠状动脉前降支结扎的方法构建缺血-再灌注小鼠模型。利用超声心动图检测各组小鼠心功能;Masson染色检测各组小鼠心脏病理学变化并测定胶原容积分数(Collagen volume fraction,CVF);蛋白免疫印迹(Western blot)法检测β-连环蛋白(β-catenin)、α-平滑肌肌动蛋白(alpha-smooth muscle actin,α-SMA)和E-钙黏蛋白(E-cadherin)的表达情况。结果与Control组比较,I/R组小鼠LVEDD和LVEDV明显增高[LVEDD(mm):3.37±0.25 vs. 4.28±0.56,LVEDV(mm):46.89±8.20 vs. 83.80±22.48],LVFS和LVEF明显降低[(LVFS:(%) 30.33±3.18 vs. 20.71±2.82,LVEF(%):58.77±4.55vs. 42.44±4.80],CVF升高(%:4.06±0.74 vs. 29.54±1.32),β-catenin及α-SMA表达明显增加(β-catenin:0.35±0.17 vs. 0.74±0.14,α-SMA:0.39±0.21 vs. 0.85±0.30),Ecadherin表达减少(0.54±0.16 vs. 0.28±0.10),差异均有统计学意义(P<0.05);与I/R组比较,I/R+TMEM88组小鼠LVEDD和LVEDV减低[LVEDD(mm):4.28±0.56 vs. 3.85±0.29,LVEDV(mm):83.80±22.48 vs. 64.27±11.59],LVFS和LVEF增高[LVFS(%):20.71±2.82vs. 24.14±3.79,LVEF(%):42.44±4.80 vs. 48.53±6.14),CVF降低(%:29.54±1.32 vs.25.06±3.62),β-catenin及α-SMA表达减少(β-catenin:0.74±0.14 vs. 0.58±0.10,α-SMA:0.85±0.30 vs. 0.59±0.11),E-cadherin表达增加(0.28±0.10 vs. 0.41±0.05),差异均有统计学意义(P<0.05)。结论 TMEM88能够通过抑制Wnt/β-catenin信号通路活性改善缺血-再灌注小鼠的心肌纤维化程度。
