靶向抑制 μ-钙蛋白酶和半胱天冬酶 9 的表达及其对韩牛卫星细胞中半胱天冬酶 3 和半胱天冬酶 7 的影响。
Targeted suppression of μ-calpain and caspase 9 expression and its effect on caspase 3 and caspase 7 in satellite cells of Korean Hanwoo cattle.
机构信息
Department of Animal Science and Institute of Rare Earth for Biological Applications, Chonbuk National University, Jeonju, 561756, Korea.
出版信息
Cell Biol Int. 2012 Sep;36(9):843-9. doi: 10.1042/CBI20120050.
The calpains play an important role in cell death and cell signalling. Caspases catalyse wholesale destruction of cellular proteins which is a major cause of cellular death. The current study looks at the function of μ-calpain and caspase 9, using RNAi (RNA interference)-mediated silencing, and to observe the mRNA expression level of caspase genes during satellite cell growth. The satellite cells were treated with siRNA (small interfering RNA) of μ-calpain and caspase 9 separately. There was reduction of 16 and 24% in CAPN1 (calpain1)-siRNA2 and CAPN1-siRNA3 transfected cells respectively, whereas it was 60 and 56% in CAPN1-siRNA1 and CAPN1-siRNA4 transfected cells respectively. CAPN1-siRNA4 and CAPN1-siRNA1 treated cells showed more reduction in caspase 3 and 7 gene expression. CARD9 (caspase recruitment domain 9)-siRNA1 and CARD9-siRNA2-treated cells showed reduction of 40 and 49% respectively. CARD9-siRNA1 and CARD9-siRNA2 showed an increase in caspase 3 gene expression, whereas CARD9-siRNA2 showed reduction in caspase 7 gene expression. These results suggest a strong cross-talk between μ-calpain and the caspase enzyme systems. Suppression of target genes, such as μ-calpain and caspase 9, might have genuine potential in the treatment of skeletal muscle atrophy.
钙蛋白酶在细胞死亡和细胞信号转导中发挥重要作用。半胱天冬酶催化细胞蛋白质的大规模破坏，这是细胞死亡的主要原因。本研究使用 RNAi（RNA 干扰）介导的沉默来研究 μ-钙蛋白酶和半胱天冬酶 9 的功能，并观察卫星细胞生长过程中半胱天冬酶基因的 mRNA 表达水平。将卫星细胞分别用 μ-钙蛋白酶和半胱天冬酶 9 的 siRNA（小干扰 RNA）处理。CAPN1-siRNA2 和 CAPN1-siRNA3 转染细胞中 CAPN1（钙蛋白酶 1）的表达分别减少了 16%和 24%，而 CAPN1-siRNA1 和 CAPN1-siRNA4 转染细胞中 CAPN1 的表达分别减少了 60%和 56%。CAPN1-siRNA4 和 CAPN1-siRNA1 处理的细胞中 caspase3 和 caspase7 基因的表达减少更为明显。CARD9-siRNA1 和 CARD9-siRNA2 处理的细胞中分别减少了 40%和 49%。CARD9-siRNA1 和 CARD9-siRNA2 使 caspase3 基因的表达增加，而 CARD9-siRNA2 使 caspase7 基因的表达减少。这些结果表明 μ-钙蛋白酶和半胱天冬酶酶系统之间存在很强的相互作用。抑制靶基因，如 μ-钙蛋白酶和半胱天冬酶 9，可能在治疗骨骼肌萎缩方面具有真正的潜力。
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