核糖体蛋白 L22表达沉默对细胞周期蛋白D1表达的影响

摘要

目的：研究核糖体蛋白L22（RPL22）基因表达沉默对细胞周期蛋白D1表达的影响。方法体外培养人肺动脉平滑肌细胞（PASMC），分为空白对照组（A组）、内皮素1（ET‐1）诱导组（B组）、siRNA‐RPL22转染组（C组）、siRNA‐RPL22＋ET‐1组（D组）、siRNA转染组（E组）和siRNA＋ET‐1组（F组）。采用RT‐PCR和Western blot分别检测RPL22、细胞周期蛋白D1的mRNA和蛋白表达，细胞计数试剂盒检测PASMC增殖情况。结果与A组和E组相比，C组RPL22、细胞周期蛋白D1 mRNA和蛋白表达均降低，PASMC增殖减少（P＜0．05）。经ET‐1诱导后，D组细胞周期蛋白D1 mRNA和蛋白表达较B组和F组降低（P＜0．05）。结论 RPL22表达沉默能有效抑制PASMC增殖，可能与下调细胞周期蛋白D1表达有关。%Objective To investigate the effect of ribosomal protein L22(RPL22) silencing on cyclin D1 expression .Methods Human pulmonary arterial smooth muscle cells (PASMC ) were cultured in vitro and then divided into six groups of A(blank control) ,B(induced by endothelin‐1) , C(transfected with siRNA‐RPL22 ) ,D (transfected with siRNA‐RPL22 followed by endothelin‐1 induction) ,E (transfected with siRNA ) and F (transfected with siRNA followed by endothelin‐1 induction) .The mRNA and protein expressions of RPL22 and cyclin D1 were detected by RT‐PCR and Western blot ,respectively .PASMC proliferation was determined by CCK‐8 assay .Results Compared with groups of A and E ,the mRNA and protein expressions of RPL22 and cyclin D1 were all significantly down‐regulated ,and PASMC proliferation was decreased in group C(P<0 .05) .The mRNA and protein expressions of cyclin D1 after endothelin‐1 induction in group D were lower than those in groups of B and F(P<0 .05) .Conclusion RPL22 silencing can effectively inhibit PASMC proliferation ,which may be related with the down‐regulation of cyclin D1 expression .