CAJ | 학술논문

目的探讨小干扰 RNAs(siRNAs)沉默 E673K-hERG 突变基因的效果及作用机制.方法 Western blot 筛选特异的 E637K-hERG 负显性突变基因靶向 siRNAs,全细胞膜片钳技术检测 siRNAs 干扰前后电流的变化.结果特异的 siRNAs 作用 E637K-hERG 后,蛋白质条带明显下调.而 WT-hERG 蛋白质条带无变化. siRNAs 干扰 WT /E637K-hERG 后,不仅使激活电流和尾电流的电流幅度都较干扰前明显增大,而且使激活电流和稳态失活电流的电向分别向正向移动了9.62mV 和17.41mV,减慢了通道电流失活速度.而 siRNAs 干扰 WT-hERG 前后电流幅度和特性无明显变化.结论 siRNAs 能有效抑制 E637K-hERG 的蛋白表达和改变 WT / E637K-hERG 的通道电流特性.
목적탐토소간우 RNAs(siRNAs)침묵 E673K-hERG 돌변기인적효과급작용궤제.방법 Western blot 사선특이적 E637K-hERG 부현성돌변기인파향 siRNAs,전세포막편겸기술검측 siRNAs 간우전후전류적변화.결과특이적 siRNAs 작용 E637K-hERG 후,단백질조대명현하조.이 WT-hERG 단백질조대무변화. siRNAs 간우 WT /E637K-hERG 후,불부사격활전류화미전류적전류폭도도교간우전명현증대,이차사격활전류화은태실활전류적전향분별향정향이동료9.62mV 화17.41mV,감만료통도전류실활속도.이 siRNAs 간우 WT-hERG 전후전류폭도화특성무명현변화.결론 siRNAs 능유효억제 E637K-hERG 적단백표체화개변 WT / E637K-hERG 적통도전류특성.
Objective To investigate the effect of short interfering RNAs (siRNAs) on silencing of E637K-hERG mu-tant gene and its mechanisms. Methods Western blot was used to screen specific siRNAs targeting dominant-nega-tive mutant of E637K-hERG. Whole-cel patch-clamp technique was used to examine the changes of electrical current before and after siRNA transfection. Results siRNA significantly down-regulated the expression of protein bands by interference of E637K-hERG, but not by WT-hERG. Through inhibition of WT/E637K-hERG but not WT-hERG, siRNA in-creased activation current and tail current amplitudes, shifted the activation curve and steady-state inactivation curve to-ward positive potentials by 9.62mV and 17.41mV, respectively, and reduced the velocity of channel inactivation. Conclu-sion siRNA can effectively inhibit E637K-hERG protein expression and alter the properties of WT/E637K-hERG channel current.