特异性核酶对肝星状细胞细胞周期蛋白D1基因表达及激活的影响  

The effect of cydin D1 specific ribozyme on the expression and activation of cydin D1 in hepatic stellate cells

作　　者：夏金荣[1] 刘乃丰[1] 黄晓明[1] 胡向阳[1] 

机构地区：[1]东南大学附属中大医院消化科,南京210009

出　　处：《中华消化杂志》2007年第3期174-177,共4页Chinese Journal of Digestion

摘　　要：目的研究抗细胞周期蛋白D1核酶对大鼠肝星状细胞（HSC）细胞周期蛋白D1基因表达及激活的影响。方法构建抗细胞周期蛋白D1核酶的真核表达载体，将其转染人HSC-T6细胞，同时以空白及转染pLXSN作为对照，用G418筛选出阳性细胞克隆；分别用RT-PCR和Western印迹法检测细胞周期蛋白D1和α-平滑肌肌动蛋白（α-SMA）的表达。结果转染核酶832的HSC细胞周期蛋白D1 mRNA和蛋白表达均受到明显抑制，分别为对照组的42．22％（P〈0．01）和58．29％（P〈0．01），同时α-SMA mRNA和蛋白表达与对照组相比也显著下降（P值均〈0．01）。结论载体表达的特异性核酶能有效地抑制HSC的细胞周期蛋白D1的表达和激活。Objectives To investigate the effect of cyclin D1 specific ribozyme on cyclin D1 expression and activation in rats hepatic stellate cells（HSCs）. Methods Expression vector of anti-cyclin D1 ribozyme was contructed and transfected into HSC-T6 cell line with lipofectin. The blank and cells transfected with pLXSN were served as controls. The positive cell clones were collected by G418 screening. The expression levels of cyclin D1 mRNA and α-smooth muscle actin were determined by reverse transcription polymerase chain reaction and Western blot. Results The ribozyme832 strongly inhibited the mRNA and protein expressions of cyclin D1 in HSCs by 42.22% and 58.29% compared to those in pLXSN control, respectively（P 〈 0. 01）. The mRNA and protein expressions of α-SMA were also reduced by 58. 29% （P 〈0.01） and 54. 56%（P（0.01） compared with those in blank control. Conclusion Ribozyme targeting cyclin D1 may effectively inhibit cyclin D1 expression and activation in the HSC T6 cell line.

关 键 词：细胞周期蛋白D1 肝细胞 基因表达 

分 类 号：R686[医药卫生—骨科学]