Aim: To evaluate the Rap1A mRNA expression and its significance in the testes of normal and azoospermic subjects. Methods: A cDNA microarray that contained Rap1A and some other genes such as RBM, EIF1 AY was used to i...Aim: To evaluate the Rap1A mRNA expression and its significance in the testes of normal and azoospermic subjects. Methods: A cDNA microarray that contained Rap1A and some other genes such as RBM, EIF1 AY was used to identify the differential gene expression profiles between the normal and azoospermic testes. cDNA probes were prepared by labeling mRNA from azoospermic and normal testicular tissues through reverse transcription with Cy5-dUTP and Cy3-dUTP, respectively. The mixed cDNA probes were then hybridized with cDNA microarray (each containing 4096 unique human cDNA sequences). The fluorescent signals were scanned and the values of Cy5-dUTP and Cy3-dUTP on each spot were analyzed and calculated. In situ hybridization was employed to detect the expression of RaplA in the testes of 10 fertile and 39 azoospermic subjects. Results: One hundred and twenty-eight differentially expressed genes were found to be possibly related to azoospermia, of which 56 were up-regulated and 72, down-regulated genes. The mRNA expression of RaplA in the spermatogenic cells of azoospermic was stronger than that in those of the fertile testes. Conclusion: Rap1A may play certain roles in the development of azoospermia.展开更多
Rap1A is a small G protein implicated in a spectrum of biological processes such as cell proliferation,adhesion,differentiation,and embryogenesis.The downstream effectors through which Rap1A mediates its diverse effec...Rap1A is a small G protein implicated in a spectrum of biological processes such as cell proliferation,adhesion,differentiation,and embryogenesis.The downstream effectors through which Rap1A mediates its diverse effects are largely unknown.Here we show that Rap1A,but not the related small G proteins Rap2 or Ras,binds the tumor suppressor Ras association domain family 1A(RASSF1A)in a manner that is regulated by phosphorylation of RASSF1A.Interaction with Rap1A is shown to influence the effect of RASSF1A on microtubule behavior.展开更多
目的探讨鸟嘌呤核苷酸交换因子C3G/Rap1酶和鸟嘌呤核苷酸交换因子Dock180/Rac1酶信号通路在卵巢癌浸润中的可能作用。方法 Western blot检测Dock180沉默的卵巢癌细胞SKOV3中C3G的表达,验证上皮性卵巢癌组织中Dock180与C3G的表达相关性;...目的探讨鸟嘌呤核苷酸交换因子C3G/Rap1酶和鸟嘌呤核苷酸交换因子Dock180/Rac1酶信号通路在卵巢癌浸润中的可能作用。方法 Western blot检测Dock180沉默的卵巢癌细胞SKOV3中C3G的表达,验证上皮性卵巢癌组织中Dock180与C3G的表达相关性;免疫组化比较卵巢癌组织中Dock180与C3G的表达趋势;免疫荧光观察SKOV3中Dock180与C3G及它们各自的下游蛋白Rac1/Rap1的定位。结果 Dock180基因沉默的细胞中C3G表达明显增强(P<0.05);Dock180与C3G在卵巢癌组织中的表达呈现相反趋势(P<0.05);C3G/Dock180均主要分布于细胞质,下游效应蛋白Rap1/Rac1在细胞膜和细胞质都有表达,但Rap1以细胞质为主,而Rac1可以伸展至细胞膜及细胞膜皱褶。结论卵巢癌细胞和组织中C3G与Dock180表达呈相反趋势,下游蛋白Rap1与Rac1在细胞内的定位分布差异,可能与C3G/Rap1和Dock180/Rac1信号通路在卵巢肿瘤浸润中的不同作用有关。展开更多
文摘Aim: To evaluate the Rap1A mRNA expression and its significance in the testes of normal and azoospermic subjects. Methods: A cDNA microarray that contained Rap1A and some other genes such as RBM, EIF1 AY was used to identify the differential gene expression profiles between the normal and azoospermic testes. cDNA probes were prepared by labeling mRNA from azoospermic and normal testicular tissues through reverse transcription with Cy5-dUTP and Cy3-dUTP, respectively. The mixed cDNA probes were then hybridized with cDNA microarray (each containing 4096 unique human cDNA sequences). The fluorescent signals were scanned and the values of Cy5-dUTP and Cy3-dUTP on each spot were analyzed and calculated. In situ hybridization was employed to detect the expression of RaplA in the testes of 10 fertile and 39 azoospermic subjects. Results: One hundred and twenty-eight differentially expressed genes were found to be possibly related to azoospermia, of which 56 were up-regulated and 72, down-regulated genes. The mRNA expression of RaplA in the spermatogenic cells of azoospermic was stronger than that in those of the fertile testes. Conclusion: Rap1A may play certain roles in the development of azoospermia.
基金SKV also acknowledges the award of the Commonwealth Scholarship and the financial support received from the Department of Medical Oncology,Medical Sciences Division,The University of Oxford,Oxford,UK and the Cancer Research UK.
文摘Rap1A is a small G protein implicated in a spectrum of biological processes such as cell proliferation,adhesion,differentiation,and embryogenesis.The downstream effectors through which Rap1A mediates its diverse effects are largely unknown.Here we show that Rap1A,but not the related small G proteins Rap2 or Ras,binds the tumor suppressor Ras association domain family 1A(RASSF1A)in a manner that is regulated by phosphorylation of RASSF1A.Interaction with Rap1A is shown to influence the effect of RASSF1A on microtubule behavior.
文摘目的探讨鸟嘌呤核苷酸交换因子C3G/Rap1酶和鸟嘌呤核苷酸交换因子Dock180/Rac1酶信号通路在卵巢癌浸润中的可能作用。方法 Western blot检测Dock180沉默的卵巢癌细胞SKOV3中C3G的表达,验证上皮性卵巢癌组织中Dock180与C3G的表达相关性;免疫组化比较卵巢癌组织中Dock180与C3G的表达趋势;免疫荧光观察SKOV3中Dock180与C3G及它们各自的下游蛋白Rac1/Rap1的定位。结果 Dock180基因沉默的细胞中C3G表达明显增强(P<0.05);Dock180与C3G在卵巢癌组织中的表达呈现相反趋势(P<0.05);C3G/Dock180均主要分布于细胞质,下游效应蛋白Rap1/Rac1在细胞膜和细胞质都有表达,但Rap1以细胞质为主,而Rac1可以伸展至细胞膜及细胞膜皱褶。结论卵巢癌细胞和组织中C3G与Dock180表达呈相反趋势,下游蛋白Rap1与Rac1在细胞内的定位分布差异,可能与C3G/Rap1和Dock180/Rac1信号通路在卵巢肿瘤浸润中的不同作用有关。
