目的:探讨人黑素瘤抗原-A(melanoma antigen-A,MAGE-A)对p53转录活性的影响。方法:选用p53缺失型人肺癌细胞系H1299,采用荧光素酶报告分析法、RT-PCR、Western印迹法、克隆形成实验和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(termin...目的:探讨人黑素瘤抗原-A(melanoma antigen-A,MAGE-A)对p53转录活性的影响。方法:选用p53缺失型人肺癌细胞系H1299,采用荧光素酶报告分析法、RT-PCR、Western印迹法、克隆形成实验和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling,TUNEL)等方法研究MAGE-A4对p53转录活性的影响。结果:荧光素酶报告分析结果显示,转染p53基因(25ng)可以使p21WAF1启动子介导的荧光素酶表达增加(P<0.01);共转染恒定量(25ng)的p53基因和不同量的MAGE-A4基因(100、200和400ng)后,p21WAF1启动子介导的荧光素酶表达均明显高于单独转染p53基因时的表达量(P<0.01)。RT-PCR和Western印迹法检测结果也显示,转染p53基因可以使p21WAF1mRNA和蛋白表达水平增加(P<0.01),共转染MAGE-A4基因和p53基因后,p21WAF1mRNA和蛋白表达水平均明显高于单独转染p53基因时的表达水平(P<0.01)。克隆形成实验及TUNEL染色结果显示,转染p53基因可以使H1299细胞克隆形成数减少及凋亡细胞数增加(P<0.01),共转染MAGE-A4基因和p53基因以后,H1299细胞克隆形成数与单独转染p53基因组相比减少,而凋亡细胞数与单独转染p53基因组比较增加(P<0.01)。结论:MAGE-A4可以增强p53的转录活性及功能发挥。展开更多
目的检测人脑膜瘤组织中黑色素瘤相关抗原MAGE-D4 m RNA的表达并分析其临床意义。方法采用逆转录-聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)方法,检测36例脑膜瘤组织和8例正常人脑组织中MAGE-D4 m RNA的表...目的检测人脑膜瘤组织中黑色素瘤相关抗原MAGE-D4 m RNA的表达并分析其临床意义。方法采用逆转录-聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)方法,检测36例脑膜瘤组织和8例正常人脑组织中MAGE-D4 m RNA的表达,分析MAGE-D4 m RNA的表达与脑膜瘤患者临床病理特征的关系。结果 MAGE-D4 m RNA在脑膜瘤组织中的阳性表达率为58.33%(21/36),明显高于正常人脑组织中的阳性表达率12.50%(1/8)(P<0.05);脑膜瘤组织中MAGE-D4 m RNA的表达与性别、年龄、肿瘤大小和病理类型等临床病理特征无关(P>0.05)。结论在脑膜瘤组织中MAGE-D4的表达率较高,提示MAGE-D4可能成为脑膜瘤免疫治疗的靶抗原。展开更多
MAGED4B belongs to the melanoma-associated antigen family;originally found in melanoma,it is expressed in various types of cancer,and is especially enriched in glioblastoma.However,the functional role and molecular me...MAGED4B belongs to the melanoma-associated antigen family;originally found in melanoma,it is expressed in various types of cancer,and is especially enriched in glioblastoma.However,the functional role and molecular mechanisms of MAGED4B in glioma are still unclear.In this study,we found that the MAGED4B level was higher in glioma tissue than that in non-cancer tissue,and the level was positively correlated with glioma grade,tumor diameter,Ki-67 level,and patient age.The patients with higher levels had a worse prognosis than those with lower MAGED4B levels.In glioma cells,MAGED4B overexpression promoted proliferation,invasion,and migration,as well as decreasing apoptosis and the chemosensitivity to cisplatin and temozolomide.On the contrary,MAGED4B knockdown in glioma cells inhibited proliferation,invasion,and migration,as well as increasing apoptosis and the chemosensitivity to cisplatin and temozolomide.MAGED4B knockdown also inhibited the growth of gliomas implanted into the rat brain.The interaction between MAGED4B and tripartite motif-containing 27(TRIM27)in glioma cells was detected by co-immunoprecipitation assay,which showed that MAGED4B was co-localized with TRIM27.In addition,MAGED4B overexpression down-regulated the TRIM27 protein level,and this was blocked by carbobenzoxyl-L-leucyl-L-leucyl-L-leucine(MG132),an inhibitor of the proteasome.On the contrary,MAGED4B knockdown up-regulated the TRIM27 level.Furthermore,MAGED4B overexpression increased TRIM27 ubiquitination in the presence of MG132.Accordingly,MAGED4B down-regulated the protein levels of genes downstream of ubiquitin-specific protease 7(USP7)involved in the tumor necrosis factor-alpha(TNF-α)-induced apoptotic pathway.These findings indicate that MAGED4B promotes glioma growth via a TRIM27/USP7/receptor-interacting serine/threonine-protein kinase 1(RIP1)-dependent TNF-α-induced apoptotic pathway,which suggests that MAGED4B is a potential target for glioma diagnosis and treatment.展开更多
文摘目的:探讨人黑素瘤抗原-A(melanoma antigen-A,MAGE-A)对p53转录活性的影响。方法:选用p53缺失型人肺癌细胞系H1299,采用荧光素酶报告分析法、RT-PCR、Western印迹法、克隆形成实验和末端脱氧核苷酸转移酶介导的dUTP缺口末端标记(terminal deoxynucleotidyl transferase biotin-dUTP nick end labeling,TUNEL)等方法研究MAGE-A4对p53转录活性的影响。结果:荧光素酶报告分析结果显示,转染p53基因(25ng)可以使p21WAF1启动子介导的荧光素酶表达增加(P<0.01);共转染恒定量(25ng)的p53基因和不同量的MAGE-A4基因(100、200和400ng)后,p21WAF1启动子介导的荧光素酶表达均明显高于单独转染p53基因时的表达量(P<0.01)。RT-PCR和Western印迹法检测结果也显示,转染p53基因可以使p21WAF1mRNA和蛋白表达水平增加(P<0.01),共转染MAGE-A4基因和p53基因后,p21WAF1mRNA和蛋白表达水平均明显高于单独转染p53基因时的表达水平(P<0.01)。克隆形成实验及TUNEL染色结果显示,转染p53基因可以使H1299细胞克隆形成数减少及凋亡细胞数增加(P<0.01),共转染MAGE-A4基因和p53基因以后,H1299细胞克隆形成数与单独转染p53基因组相比减少,而凋亡细胞数与单独转染p53基因组比较增加(P<0.01)。结论:MAGE-A4可以增强p53的转录活性及功能发挥。
文摘目的检测人脑膜瘤组织中黑色素瘤相关抗原MAGE-D4 m RNA的表达并分析其临床意义。方法采用逆转录-聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)方法,检测36例脑膜瘤组织和8例正常人脑组织中MAGE-D4 m RNA的表达,分析MAGE-D4 m RNA的表达与脑膜瘤患者临床病理特征的关系。结果 MAGE-D4 m RNA在脑膜瘤组织中的阳性表达率为58.33%(21/36),明显高于正常人脑组织中的阳性表达率12.50%(1/8)(P<0.05);脑膜瘤组织中MAGE-D4 m RNA的表达与性别、年龄、肿瘤大小和病理类型等临床病理特征无关(P>0.05)。结论在脑膜瘤组织中MAGE-D4的表达率较高,提示MAGE-D4可能成为脑膜瘤免疫治疗的靶抗原。
基金supported by the National Natural Science Foundation of China(81801679,81571308).
文摘MAGED4B belongs to the melanoma-associated antigen family;originally found in melanoma,it is expressed in various types of cancer,and is especially enriched in glioblastoma.However,the functional role and molecular mechanisms of MAGED4B in glioma are still unclear.In this study,we found that the MAGED4B level was higher in glioma tissue than that in non-cancer tissue,and the level was positively correlated with glioma grade,tumor diameter,Ki-67 level,and patient age.The patients with higher levels had a worse prognosis than those with lower MAGED4B levels.In glioma cells,MAGED4B overexpression promoted proliferation,invasion,and migration,as well as decreasing apoptosis and the chemosensitivity to cisplatin and temozolomide.On the contrary,MAGED4B knockdown in glioma cells inhibited proliferation,invasion,and migration,as well as increasing apoptosis and the chemosensitivity to cisplatin and temozolomide.MAGED4B knockdown also inhibited the growth of gliomas implanted into the rat brain.The interaction between MAGED4B and tripartite motif-containing 27(TRIM27)in glioma cells was detected by co-immunoprecipitation assay,which showed that MAGED4B was co-localized with TRIM27.In addition,MAGED4B overexpression down-regulated the TRIM27 protein level,and this was blocked by carbobenzoxyl-L-leucyl-L-leucyl-L-leucine(MG132),an inhibitor of the proteasome.On the contrary,MAGED4B knockdown up-regulated the TRIM27 level.Furthermore,MAGED4B overexpression increased TRIM27 ubiquitination in the presence of MG132.Accordingly,MAGED4B down-regulated the protein levels of genes downstream of ubiquitin-specific protease 7(USP7)involved in the tumor necrosis factor-alpha(TNF-α)-induced apoptotic pathway.These findings indicate that MAGED4B promotes glioma growth via a TRIM27/USP7/receptor-interacting serine/threonine-protein kinase 1(RIP1)-dependent TNF-α-induced apoptotic pathway,which suggests that MAGED4B is a potential target for glioma diagnosis and treatment.
