Yao Hai-lan, Xiao Zong-hui, Ren Hong-yan, Han Ji-sheng, Liu Zhe-wei
Capital Institute of Paediatrics, Beijing 100020, China.
Bing Du Xue Bao. 2007 Jul;23(4):276-81.
To study the inhibitory effect and function characteristics of small interfering RNA (siRNA) on cosxackievirus B3(CVB3) infection by RNA interference technique, siRNA-2B against the viral 2B region was synthesized and transfected into HeLa cell, which was then infected with CVB3. The efficiency of siRNA transfection was examined by FCM, the cell toxicity of siRNA-2B by MTT, and the antiviral ability of siRNA-2B by cytopathic effect (CPE), plaque reduction assay and RT-PCR. The results showed that siRNA-2B could be transfected efficiently into HeLa cell and lasted at least 48h. High concentration of siRNA-2B didn't show any sign of toxicity to cells. siRNA-2B exhibited a significant protective effect on cell viability by specific inhibition of viral replication. It showed a close relationship between the concentrations of siRNA-2B and the antiviral effects. siRNA-2B led to dramatical reduction of viral titers in supernatant of cell culture and weakened the reinfection ability of the virus. These data proposed that siRNA-2B, targeting 2B protein, can effectively inhibit CVB3 infection in HeLa cell and exhibits its transfection efficiency, viral inhibition specificity and adose-dependant manner, suggesting its potential role in prevention and treatement of CVB3 infection.
采用RNA干扰技术研究小干扰RNA（siRNA）对柯萨奇病毒B3（CVB3）感染的抑制作用及功能特性，合成针对病毒2B区的siRNA-2B并转染至HeLa细胞，随后用CVB3感染该细胞。通过流式细胞术检测siRNA转染效率，用MTT法检测siRNA-2B的细胞毒性，用细胞病变效应（CPE）、蚀斑减少试验和RT-PCR检测siRNA-2B的抗病毒能力。结果显示，siRNA-2B能高效转染至HeLa细胞且至少持续48小时。高浓度的siRNA-2B对细胞未表现出任何毒性迹象。siRNA-2B通过特异性抑制病毒复制对细胞活力具有显著的保护作用。其显示出siRNA-2B浓度与抗病毒效果之间存在密切关系。siRNA-2B导致细胞培养上清中病毒滴度显著降低，并削弱了病毒的再感染能力。这些数据表明，靶向2B蛋白的siRNA-2B能有效抑制HeLa细胞中的CVB3感染，并表现出其转染效率、病毒抑制特异性及剂量依赖性，提示其在预防和治疗CVB3感染中的潜在作用。
