目的探讨CADM1在大肠癌细胞系中的表达及与启动子甲基化的关系。方法用RT-PCR法检测大肠癌细胞系中CADM1基因m RNA表达水平,Western blot法检测细胞系CADM1蛋白表达情况,BSP法和MSP法检测CADM1基因启动子甲基化状态。结果 CADM1 m RNA在...目的探讨CADM1在大肠癌细胞系中的表达及与启动子甲基化的关系。方法用RT-PCR法检测大肠癌细胞系中CADM1基因m RNA表达水平,Western blot法检测细胞系CADM1蛋白表达情况,BSP法和MSP法检测CADM1基因启动子甲基化状态。结果 CADM1 m RNA在24%大肠癌细胞系表达缺失,在50%细胞系中CADM1 m RNA表达减少。75%大肠癌细胞系检测到CADM1基因启动子甲基化阳性,且表达缺失。结论 CADM1基因启动子甲基化与大肠癌的发生发展有密切关系,该基因的甲基化检测可作为癌症早期筛选、监测预后的肿瘤标志物。展开更多
A tumor suppressor gene, CADM1, encoding an immunoglobulin superfamily cell adhesion molecule, is inactivated in various cancers, including non-small-cell lung cancer (NSCLC). Although promoter methylation is one of t...A tumor suppressor gene, CADM1, encoding an immunoglobulin superfamily cell adhesion molecule, is inactivated in various cancers, including non-small-cell lung cancer (NSCLC). Although promoter methylation is one of the mechanisms to suppress CADM1 expression, about half of tumors lacking CADM1 expression do not show methylation of the gene promoter. We herein investigated the possible involvement of microRNA (miRNA) in the down-regulation of CADM1. Using computational algorithms, miR-214 and miR-375 were identified as candidate miRNAs targeting CADM1. A luciferase reporter assay demonstrated that miR-214 and miR-375 repressed the promoter activity through 3’-UTR of CADM1. Quantitative RT-PCR analysis demonstrated that miR-214 and miR-375 was highly expressed in 21 (62%) and 17 cases (50%) of 34 primary NSCLCs. Notably, increased expression of miR-214 was preferentially observed in tumors with advanced pathological stages and in those lacking CADM1 expression but were not associated with the promoter methylation, suggesting that miR-214-mediated silencing would be another mechanism to suppress CADM1 expression. On the other hand, introduction of miR-214 or miR-375 into NSCLC cells decreased CADM1 protein expression. Furthermore, overexpression of miR-214 enhanced anchorage-independent growth of NSCLC cells, A549, whereas transfection of miRNA inhibitor, miR-214 or miR-375, significantly suppressed the in vitro wound healing activity of HCC827 cells. These findings suggest that overexpression of miR-214 and miR-375 could participate in the malignant features of NSCLC through down-regulating CADM1 and would provide a potential target for the treatment of a subset of NSCLC.展开更多
BACKGROUND： Increasing evidence indicates that down- regulation of cell adhesion molecule 1 （CADM1） contributes to tumorigenesis in various cancers. The present study was undertaken to investigate the CADM1 express...BACKGROUND： Increasing evidence indicates that down- regulation of cell adhesion molecule 1 （CADM1） contributes to tumorigenesis in various cancers. The present study was undertaken to investigate the CADM1 expression pattern in human hepatocellular carcinoma （HCC）, and to elucidate the mechanism underlying CADMl-mediated tumor suppression.展开更多
目的:研究过表达细胞黏附分子1(cell adhesion molecule 1,CADM1)对卵巢癌增殖、迁移和侵袭的影响。方法:q RT-PCR测定CADM1m RNA在卵巢癌细胞系SKOV3及人正常卵巢上皮细胞hose中的表达,将SKOV3细胞分成两组,即CADM1过表达组和对照组,转...目的:研究过表达细胞黏附分子1(cell adhesion molecule 1,CADM1)对卵巢癌增殖、迁移和侵袭的影响。方法:q RT-PCR测定CADM1m RNA在卵巢癌细胞系SKOV3及人正常卵巢上皮细胞hose中的表达,将SKOV3细胞分成两组,即CADM1过表达组和对照组,转染48 h后Western印迹测定两组CADM1蛋白表达量,采用lipofectamine 2000分别转染pc DNA3.1-CADM1及pc DNA3.1质粒,采用CCK-8、克隆形成、细胞划痕及Transwel l实验分别检测两组细胞增殖、克隆形成、细胞迁移及侵袭能力。结果:CADM1m RNA在SKOV3中表达水平显著低于hose细胞系(1.54±0.34 vs.5.63±0.96,P<0.0 5);转染4 8 h后,C A D M 1过表达组和对照组C A D M 1蛋白表达量分别为2.5 3±0.4 2,0.37±0.09,差异具有统计学意义(P<0.05)。CADM1过表达组和对照组在0,24,48,72 h 450 nm处的OD值差异无统计学意义(P>0.05);CADM1过表达组与对照组克隆形成数相比(60.4±7.6 vs.58.3±8.2),差异无统计学意义(P>0.05);CADM1过表达组细胞迁移率显著低于对照组(20.3%±3.5%v s.60.1%±4.2%,P<0.05);C A DM1过表达组侵袭细胞数显著少于对照组(24.5±5.3 v s.65.1±6.9,P<0.05)。结论:CADM1在卵巢癌细胞系中低表达,过表达CADM1对卵巢癌细胞增殖和克隆形成无影响,但可抑制迁移和侵袭,起抑癌基因的作用。展开更多
文摘目的探讨CADM1在大肠癌细胞系中的表达及与启动子甲基化的关系。方法用RT-PCR法检测大肠癌细胞系中CADM1基因m RNA表达水平,Western blot法检测细胞系CADM1蛋白表达情况,BSP法和MSP法检测CADM1基因启动子甲基化状态。结果 CADM1 m RNA在24%大肠癌细胞系表达缺失,在50%细胞系中CADM1 m RNA表达减少。75%大肠癌细胞系检测到CADM1基因启动子甲基化阳性,且表达缺失。结论 CADM1基因启动子甲基化与大肠癌的发生发展有密切关系,该基因的甲基化检测可作为癌症早期筛选、监测预后的肿瘤标志物。
文摘A tumor suppressor gene, CADM1, encoding an immunoglobulin superfamily cell adhesion molecule, is inactivated in various cancers, including non-small-cell lung cancer (NSCLC). Although promoter methylation is one of the mechanisms to suppress CADM1 expression, about half of tumors lacking CADM1 expression do not show methylation of the gene promoter. We herein investigated the possible involvement of microRNA (miRNA) in the down-regulation of CADM1. Using computational algorithms, miR-214 and miR-375 were identified as candidate miRNAs targeting CADM1. A luciferase reporter assay demonstrated that miR-214 and miR-375 repressed the promoter activity through 3’-UTR of CADM1. Quantitative RT-PCR analysis demonstrated that miR-214 and miR-375 was highly expressed in 21 (62%) and 17 cases (50%) of 34 primary NSCLCs. Notably, increased expression of miR-214 was preferentially observed in tumors with advanced pathological stages and in those lacking CADM1 expression but were not associated with the promoter methylation, suggesting that miR-214-mediated silencing would be another mechanism to suppress CADM1 expression. On the other hand, introduction of miR-214 or miR-375 into NSCLC cells decreased CADM1 protein expression. Furthermore, overexpression of miR-214 enhanced anchorage-independent growth of NSCLC cells, A549, whereas transfection of miRNA inhibitor, miR-214 or miR-375, significantly suppressed the in vitro wound healing activity of HCC827 cells. These findings suggest that overexpression of miR-214 and miR-375 could participate in the malignant features of NSCLC through down-regulating CADM1 and would provide a potential target for the treatment of a subset of NSCLC.
基金supported by grants from the National S&T Major Project(2012ZX10002-017)National Natural Science Foundation of China(81201944)+1 种基金Health Science and Technology Program of Zhejiang Province(2013KYB083)Education Department of Zhejiang Province Scientific Research Project(Y201430751)
文摘BACKGROUND： Increasing evidence indicates that down- regulation of cell adhesion molecule 1 （CADM1） contributes to tumorigenesis in various cancers. The present study was undertaken to investigate the CADM1 expression pattern in human hepatocellular carcinoma （HCC）, and to elucidate the mechanism underlying CADMl-mediated tumor suppression.
文摘目的:研究过表达细胞黏附分子1(cell adhesion molecule 1,CADM1)对卵巢癌增殖、迁移和侵袭的影响。方法:q RT-PCR测定CADM1m RNA在卵巢癌细胞系SKOV3及人正常卵巢上皮细胞hose中的表达,将SKOV3细胞分成两组,即CADM1过表达组和对照组,转染48 h后Western印迹测定两组CADM1蛋白表达量,采用lipofectamine 2000分别转染pc DNA3.1-CADM1及pc DNA3.1质粒,采用CCK-8、克隆形成、细胞划痕及Transwel l实验分别检测两组细胞增殖、克隆形成、细胞迁移及侵袭能力。结果:CADM1m RNA在SKOV3中表达水平显著低于hose细胞系(1.54±0.34 vs.5.63±0.96,P<0.0 5);转染4 8 h后,C A D M 1过表达组和对照组C A D M 1蛋白表达量分别为2.5 3±0.4 2,0.37±0.09,差异具有统计学意义(P<0.05)。CADM1过表达组和对照组在0,24,48,72 h 450 nm处的OD值差异无统计学意义(P>0.05);CADM1过表达组与对照组克隆形成数相比(60.4±7.6 vs.58.3±8.2),差异无统计学意义(P>0.05);CADM1过表达组细胞迁移率显著低于对照组(20.3%±3.5%v s.60.1%±4.2%,P<0.05);C A DM1过表达组侵袭细胞数显著少于对照组(24.5±5.3 v s.65.1±6.9,P<0.05)。结论:CADM1在卵巢癌细胞系中低表达,过表达CADM1对卵巢癌细胞增殖和克隆形成无影响,但可抑制迁移和侵袭,起抑癌基因的作用。
