腺病毒介导RNA干扰抑制核心结合因子α1表达阻断软骨细胞的肥大分化

摘要

背景：软骨细胞肥大分化是软骨内骨化启动的标志，也是软骨内骨化必不可少的步骤，它是一种级联反应，一旦启动就很难阻断，最终结果是形成骨骼结构。RNA干扰技术是一种转录后水平的基因沉默，相关研究显示采用RNA干扰技术阻断核心结合因子α1表达能够有效地抑制异位骨化形成。目的：利用RNA干扰技术抑制核心结合因子α1表达，从而阻断大鼠软骨细胞肥大分化。方法：构建沉默SD大鼠核心结合因子基因的腺病毒Ad-Cbfa1-siRNA。利用维甲酸及白细胞介素1α促进软骨细胞肥大分化，观察Ad-Cbfa1-siRNA对核心结合因子α1表达的抑制作用。利用核心结合因子α1免疫组化方法比较各组核心结合因子α1的表达从而分析软骨细胞的肥大分化情况。结果与结论：经维甲酸和白细胞介素1α诱导后，阴性对照病毒组软骨细胞出现肥大分化，核心结合因子α1的表达呈阳性反应；Ad-Cbfa1-siRNA组软骨细胞中未见核心结合因子α1明显表达。提示利用RNA干扰技术能够明显抑制核心结合因子α1的表达，从而阻断软骨细胞的肥大分化。%BACKGROUND:Hypertrophic differentiation of chondrocytes is the sign of starting endochondral ossification, and it is also an essential step in endochondral ossification, which is a cascade reaction and difficult to be blocked once started. The end result is the formation of bone structure. RNA interference is a post-transcriptional gene silencing. Relevant studies have shown that the use of RNA interference to block the expression of core binding factorα1 (Cbfα1) can effectively inhibit the formation of heterotopic ossification. OBJECTIVE:To use RNA intereference technology to suppress Cbfα1 expression so as to achieve the purpose of blocking the hypertrophic diferentiation of chondrocytes. METHODs: We constructed an adenovirus containing siRNA against Cbfα1 (Ad-Cbfα1-siRNA). Retinoic acid and interleukin-1α were used to induce hypertrophic differetiation of chondrocytes, and then Ad-Cbfα1-siRNA was utilized to inhibit the hypertrophic differentiation of chondrocytes. Immunohistochemistry method was used to analyze the expression of Cbfα1. RESULTS AND CONCLUSION:After induction with retinoic acid and interleukin-1α, the chondrocytes in the negative control virus group appeared to have hypertrophy and the expression of Cbfα1 was positive. In the Ad-Cbα1-siRNA group, the expression of Cbfα1 was negative. These findings suggest that the inhibition of Cbfα1 by RNA interference can be a powerful way to prevent the hypertrophic differentiation of chondrocytes .