AIM:To investigate the expression profile of miRNA in esophageal squamous cell carcinoma(ESCC).METHODS:The expression profile of miRNA in ESCC tissues was analyzed by miRNA microarray.The expression levels of miR-143 ...AIM:To investigate the expression profile of miRNA in esophageal squamous cell carcinoma(ESCC).METHODS:The expression profile of miRNA in ESCC tissues was analyzed by miRNA microarray.The expression levels of miR-143 and miR-145 in 86 ESCC patients were determined by real-time polymerase chain reaction(PCR) using TaqMan assay.The mobility effect was estimated by wound-healing using esophageal carcinoma cells transfected with miRNA expression plasmids.RESULTS:A set of miRNAs was found to be deregulated in the ESCC tissues,and the expression levels of miR-143 and-145 were significantly decreased in most of the ESCC tissues examined.Both miR-143 and miR-145 expression correlated with tumor invasion depth.The transfection of human esophageal carcinoma cells with miR-143 and miR-145 expression plasmids resulted in a greater inhibition of cell mobility,however,the protein level of the previously reported target of miR-145,FSCN1,did not show any significant downregulation.CONCLUSION:These findings suggest that the deregulation of miRNAs plays an important role in the progression of ESCC.Both miR-143 and miR-145 might act as anti-oncomirs common to ESCC.展开更多
目的·研究微小RNA-143(miR-143)在人结肠癌及癌周正常组织中的表达,探索其对结肠癌HCT116、RKO细胞功能的影响及下游靶基因的预测。方法·设计合成并构建miR-143慢病毒表达载体,稳定转染人结肠癌HCT116、RKO细胞。实时荧光定量...目的·研究微小RNA-143(miR-143)在人结肠癌及癌周正常组织中的表达,探索其对结肠癌HCT116、RKO细胞功能的影响及下游靶基因的预测。方法·设计合成并构建miR-143慢病毒表达载体,稳定转染人结肠癌HCT116、RKO细胞。实时荧光定量PCR检测人结肠癌、癌周正常组织、慢病毒感染后HCT116和RKO细胞中miR-143的表达。Annexin V-APC单染法及流式细胞仪检测感染前后细胞凋亡能力。Transwell法检测感染前后HCT116、RKO细胞迁移能力的改变。Western blotting方法检测FOSL2蛋白的表达。结果·miR-143在结肠癌组织中的表达量显著低于癌周正常组织(0.339±0.454 vs 1.003±0.003)(U=16.000,Z=-4.231,P=0.000)。上调miR-143后,HCT116细胞凋亡率显著高于阴性对照组(P=0.000),RKO细胞凋亡率显著高于阴性对照组(P=0.000)。上调miR-143后,HCT116迁移细胞数显著低于阴性对照组(P=0.000)。miR-143下调后,RKO迁移细胞数显著高于阴性对照组(P=0.003)。miR-143下调后,FOSL2蛋白表达量升高。结论·miR-143在结肠癌组织中表达量显著下降。上调miR-143可促进结肠癌细胞凋亡,并抑制HCT116细胞迁移;下调miR-143可促进RKO细胞迁移;抑制miR-143功能可上调FOSL2蛋白表达。展开更多
基金Supported by Grants from the NSFC-Guangdong Joint Fund,No.U0932001the National Natural Science Foundation of China,No.30900560
文摘AIM:To investigate the expression profile of miRNA in esophageal squamous cell carcinoma(ESCC).METHODS:The expression profile of miRNA in ESCC tissues was analyzed by miRNA microarray.The expression levels of miR-143 and miR-145 in 86 ESCC patients were determined by real-time polymerase chain reaction(PCR) using TaqMan assay.The mobility effect was estimated by wound-healing using esophageal carcinoma cells transfected with miRNA expression plasmids.RESULTS:A set of miRNAs was found to be deregulated in the ESCC tissues,and the expression levels of miR-143 and-145 were significantly decreased in most of the ESCC tissues examined.Both miR-143 and miR-145 expression correlated with tumor invasion depth.The transfection of human esophageal carcinoma cells with miR-143 and miR-145 expression plasmids resulted in a greater inhibition of cell mobility,however,the protein level of the previously reported target of miR-145,FSCN1,did not show any significant downregulation.CONCLUSION:These findings suggest that the deregulation of miRNAs plays an important role in the progression of ESCC.Both miR-143 and miR-145 might act as anti-oncomirs common to ESCC.
文摘目的·研究微小RNA-143(miR-143)在人结肠癌及癌周正常组织中的表达,探索其对结肠癌HCT116、RKO细胞功能的影响及下游靶基因的预测。方法·设计合成并构建miR-143慢病毒表达载体,稳定转染人结肠癌HCT116、RKO细胞。实时荧光定量PCR检测人结肠癌、癌周正常组织、慢病毒感染后HCT116和RKO细胞中miR-143的表达。Annexin V-APC单染法及流式细胞仪检测感染前后细胞凋亡能力。Transwell法检测感染前后HCT116、RKO细胞迁移能力的改变。Western blotting方法检测FOSL2蛋白的表达。结果·miR-143在结肠癌组织中的表达量显著低于癌周正常组织(0.339±0.454 vs 1.003±0.003)(U=16.000,Z=-4.231,P=0.000)。上调miR-143后,HCT116细胞凋亡率显著高于阴性对照组(P=0.000),RKO细胞凋亡率显著高于阴性对照组(P=0.000)。上调miR-143后,HCT116迁移细胞数显著低于阴性对照组(P=0.000)。miR-143下调后,RKO迁移细胞数显著高于阴性对照组(P=0.003)。miR-143下调后,FOSL2蛋白表达量升高。结论·miR-143在结肠癌组织中表达量显著下降。上调miR-143可促进结肠癌细胞凋亡,并抑制HCT116细胞迁移;下调miR-143可促进RKO细胞迁移;抑制miR-143功能可上调FOSL2蛋白表达。
