siRNA干扰Wnt11对鸭成肌细胞增殖的影响  

Effects of Wnt11 by siRNA interfering on cell proliferation of duck myoblasts

作　　者：解晓东[1] 虞德兵[1] 于敏莉[1] 林凯[1] 何宗亮 杜文兴[1] 

机构地区：[1]南京农业大学动物科技学院,江苏南京210095 [2]南京市畜牧家禽科学研究所,江苏南京210036

出　　处：《畜牧与兽医》2017年第2期42-46,共5页Animal Husbandry & Veterinary Medicine

基　　金：江苏省农业科技自主创新资金项目(CX(11)1033);国家科技支撑项目(2013BAD20B05)

摘　　要：为了探究Wnt11(无翅型MMTV整合位点家族成员11)对鸭骨骼肌成肌细胞增殖的影响以及可能与Wnt11相关的信号通路,采用体外分离培养鸭骨骼肌成肌细胞,脂质体介导siRNA(小干扰RNA)转染干扰基因的表达,qRT-PCR(实时荧光定量PCR)检测基因表达变化,Ed U(5-乙炔基-2'-脱氧尿苷)染色分析细胞增殖情况,流式细胞术检测细胞周期变化及信号通路抑制剂处理。结果显示:干扰Wnt11的表达后,增殖相关基因CCND1(细胞周期蛋白D1)、CDK6(周期蛋白依赖性激酶6)和PCNA(增殖细胞核抗原)的表达量极显著降低(P<0.01),Ed U阳性细胞数极显著降低(P<0.01),处于S期的细胞数显著降低(P<0.05);当用Akt(蛋白激酶B)抑制剂处理鸭成肌细胞后,Wnt11表达量显著降低(P<0.05),而干扰Wnt11表达后,PI3K(磷脂酰肌醇-3-羟激酶)和Akt的表达量显著升高(P<0.05)。提示:Wnt11在鸭成肌细胞增殖过程中有重要调控作用,Wnt11发挥调控作用需要PI3K/Akt信号通路参与,且二者间存在反馈作用。In order to investigate the effect of Wnt11 by siRNA interfering on cell proliferation of duck myoblast and the relevant pathway,duck myoblast were cultured in vitro.The siRNA were used to inhibit the expression of Wnt11 gene and qRT-PCR used to detect the gene expression.In addition,Ed U and immunofluorescence were used to determine the cell proliferation and MK2206 used to inhibit PI3 K/Akt pathway.The results showed that expressions of CCND1,CDK6 and PCNA were significantly decreased（P〈0.01）,amount of Ed U positive cells was significantly decreased（P〈0.01） and S phase proportion was also reduced（P〈0.05） after Wnt11 was inhibited.Moreover,levels of Wnt11 were significantly decreased after treated with MK2206（P〈0.05） and levels of PI3K and Akt significantly higher treated with siRNA-Wnt11（P〈0.05）.Together,we suggested that Wnt11 played an important role on regulating duck myoblast proliferation and PI3K/Akt signal pathway was involved in this process.There was a feedback between Wnt11 signals and PI3K/Akt signal pathway.

关 键 词：SIRNA Wnt11 鸭成肌细胞 增殖 

分 类 号：S834[农业科学—畜牧学]