This study was performed to investigate a potential marker for the presence of spermatozoa in the ejaculate following varicocelectomy in Chinese men with nonobstructive azoospermia and varicoceles. The micro-RNA （miR...This study was performed to investigate a potential marker for the presence of spermatozoa in the ejaculate following varicocelectomy in Chinese men with nonobstructive azoospermia and varicoceles. The micro-RNA （miR）-192a levels in seminal plasma and testicular tissue were evaluated by quantitative real-time polymerase chain reaction from 60 men with nonobstructive azoospermia and varicoceles （Group A： 27 men with spermatozoa found in the ejaculate after surgery; Group B： 33 men without spermatozoa found in the ejaculate after surgery） and 30 controls. The seminal plasma and testicular tissue miR-192a levels were higher in Group B than in Group A and the controls （P〈 0.001）, and there was no significant difference between Group A and the controls （P〉 0.05）. Apoptosis and proliferation assays with miR mimics and inhibitors showed that miR-192a induced GC-2 cell apoptosis through the activation of Caspase-3 protein. Thus, seminal plasma miR-192a appears to be a potential marker for successfully indicating spermatozoa in the ejaculate following microsurgical varicocelectomy in men with nonobstructive azoospermia and varicoceles. Seminal plasma miR-192a may be a useful clinical marker for prescreening to determine which patients with nonobstructive azoospermia and varicoceles would benefit from varicocelectomy.展开更多
目的探讨HBx基因通过调节miR-192的表达影响人肝癌细胞株HepG2周期进展的机制。方法流式细胞仪分析以下3组细胞的周期变化:HepG2/HBx细胞(HepG2细胞稳定转染HBx基因)、HepG2/pcDNA3.1细胞(HepG2细胞稳定转染空载体pcDNA3.1)以及HepG2细...目的探讨HBx基因通过调节miR-192的表达影响人肝癌细胞株HepG2周期进展的机制。方法流式细胞仪分析以下3组细胞的周期变化:HepG2/HBx细胞(HepG2细胞稳定转染HBx基因)、HepG2/pcDNA3.1细胞(HepG2细胞稳定转染空载体pcDNA3.1)以及HepG2细胞。3组细胞中miR-192的表达采用Taqman探针荧光定量PCR法检测。流式细胞术观察转染miR-192后HepG2细胞的周期分布变化,SYBR Green荧光定量PCR和Westernblot分别检测miR-192对HepG2细胞p53、CDKN1A mRNA和蛋白表达的影响。结果 3组细胞中,HepG2/HBx细胞G0/G1期细胞比例明显降低[(52.78±4.08)%vs(67.37±4.87)%,(65.08±5.15)%],S期和G2/M期比例明显升高[S期:(25.22±1.84)%vs(19.78±1.26)%,(18.84±1.68)%;G2/M期:(22.00±2.07)%vs(12.85±1.29)%,(16.08±1.44)%]。HepG2/HBx细胞miR-192表达显著下调[(49.1±5.9)%vs(98.0±8.9)%,(100.0±9.1)%]。转染miR-192引起HepG2细胞G0/G1期和G2/M期阻滞,同时p53、CDKN1A mRNA(p53:1.68±0.12 vs 0.90±0.06;CD-KN1A:2.36±0.12 vs 1.05±0.06)和蛋白(p53:3.07倍;CDKN1A:2.82倍)的表达水平亦显著上升。结论 miR-192通过促进p53和CDKN1A表达引起HepG2细胞周期阻滞,而HBx通过下调miR-192加速HepG2细胞周期进程。展开更多
文摘This study was performed to investigate a potential marker for the presence of spermatozoa in the ejaculate following varicocelectomy in Chinese men with nonobstructive azoospermia and varicoceles. The micro-RNA （miR）-192a levels in seminal plasma and testicular tissue were evaluated by quantitative real-time polymerase chain reaction from 60 men with nonobstructive azoospermia and varicoceles （Group A： 27 men with spermatozoa found in the ejaculate after surgery; Group B： 33 men without spermatozoa found in the ejaculate after surgery） and 30 controls. The seminal plasma and testicular tissue miR-192a levels were higher in Group B than in Group A and the controls （P〈 0.001）, and there was no significant difference between Group A and the controls （P〉 0.05）. Apoptosis and proliferation assays with miR mimics and inhibitors showed that miR-192a induced GC-2 cell apoptosis through the activation of Caspase-3 protein. Thus, seminal plasma miR-192a appears to be a potential marker for successfully indicating spermatozoa in the ejaculate following microsurgical varicocelectomy in men with nonobstructive azoospermia and varicoceles. Seminal plasma miR-192a may be a useful clinical marker for prescreening to determine which patients with nonobstructive azoospermia and varicoceles would benefit from varicocelectomy.
文摘目的探讨HBx基因通过调节miR-192的表达影响人肝癌细胞株HepG2周期进展的机制。方法流式细胞仪分析以下3组细胞的周期变化:HepG2/HBx细胞(HepG2细胞稳定转染HBx基因)、HepG2/pcDNA3.1细胞(HepG2细胞稳定转染空载体pcDNA3.1)以及HepG2细胞。3组细胞中miR-192的表达采用Taqman探针荧光定量PCR法检测。流式细胞术观察转染miR-192后HepG2细胞的周期分布变化,SYBR Green荧光定量PCR和Westernblot分别检测miR-192对HepG2细胞p53、CDKN1A mRNA和蛋白表达的影响。结果 3组细胞中,HepG2/HBx细胞G0/G1期细胞比例明显降低[(52.78±4.08)%vs(67.37±4.87)%,(65.08±5.15)%],S期和G2/M期比例明显升高[S期:(25.22±1.84)%vs(19.78±1.26)%,(18.84±1.68)%;G2/M期:(22.00±2.07)%vs(12.85±1.29)%,(16.08±1.44)%]。HepG2/HBx细胞miR-192表达显著下调[(49.1±5.9)%vs(98.0±8.9)%,(100.0±9.1)%]。转染miR-192引起HepG2细胞G0/G1期和G2/M期阻滞,同时p53、CDKN1A mRNA(p53:1.68±0.12 vs 0.90±0.06;CD-KN1A:2.36±0.12 vs 1.05±0.06)和蛋白(p53:3.07倍;CDKN1A:2.82倍)的表达水平亦显著上升。结论 miR-192通过促进p53和CDKN1A表达引起HepG2细胞周期阻滞,而HBx通过下调miR-192加速HepG2细胞周期进程。
