蛋白激酶CK2β小干扰RNA促进小鼠受精卵早期发育  

Promotion of protein kinase CK2β siRNA in early development of fertilized mouse eggs

作　　者：刘超[1] 任丽莉[1] 刘洋[1] 肖建英[1] 

机构地区：[1]辽宁医学院生物化学与分子生物学教研室,辽宁锦州121001

出　　处：《吉林大学学报（医学版）》2012年第4期678-682,共5页Journal of Jilin University：Medicine Edition

基　　金：辽宁省教育厅重点实验室项目资助课题(LS2010101);辽宁医学院青年科技启动基金资助课题(Y2011Z014)

摘　　要：目的:利用特异性小发夹RNA(siRNA)寡核苷酸片段,研究蛋白激酶CK2调节亚基CK2β对小鼠受精卵早期发育的影响,阐明调控小鼠受精卵早期发育的机制。方法:超排卵获得小鼠1-细胞期受精卵,分别显微注射TE缓冲液、scramble siRNA和CK2βsiRNA,收集卵细胞后用RT-PCR和Western blotting方法检测CK2βsiRNA的干涉效能;相差显微镜观察受精卵的形态变化、计数卵裂率;放射自显影测定M期促进因子(MPF)活性。结果:与TE缓冲液和scramble siRNA组比较,CK2βsiRNA组CK2β的mRNA水平和蛋白表达显著降低(P<0.01),MPF的活性明显升高(P<0.01),在人绒毛膜促性腺激素(hCG)注射后28.5h达到高峰;受精卵第一次有丝分裂加速,CK2βsiRNA组在hCG注射后28.5h大多数受精卵已经分裂为2-细胞期胚胎;hCG注射后32.0h卵裂率为89.5%,与TE缓冲液和scramble siRNA组比较差异有统计学意义(P<0.01)。结论:CK2βsiRNA通过提前激活MPF活性,进而促进小鼠1-细胞期受精卵的发育。Objective To study the influence of the regulatory subunit of protein kinase CK2β in the early development of fertilized mouse eggs by the means of specific siRNA and to clarify the mechanism of regulation of early development of fertilized mouse eggs.Methods One-cell stage fertilized mouse eggs were collected after superovulation,and microinjected with TE buffer,scramble siRNA or CK2β siRNA,then the effect of siRNA was examined by RT-PCR and Western blotting.After RNA interference,the morphological changes and cleavage rate of mouse embryos were observed under dissecting microscope,and the M-phase promoting factor（MPF）activity of fertilized mouse eggs were detected by autoradiography.Results Compared with TE buffer and scramble siRNA groups,the expression levels of CK2β mRNA and protein in CK2β siRNA group were significantly reduced（P〈0.01）;the activity of MPF in CK2β siRNA group reached its peak value at 28.5 h after human chorionic gonadotropin（hCG） injection,about 30 min earlier than TE buffer and scramble siRNA groups.The embryos treated with CK2β siRNA entered the M phase at 28.5 h after hCG injection,and the cleavage rate was as high as 89.5% at 32.0 h,markedly higher than those in TE buffer and scramble siRNA groups（P〈0.01）.Conclusion CK2β siRNA may promote the early development of fertilized mouse eggs via activating MPF activity ahead of time.

关 键 词：蛋白激酶CK2β SIRNA M期促进因子 小鼠受精卵 

分 类 号：Q132[生物学—普通生物学]