Department of Respiratory Medicine, The First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, P.R. China.
Int J Mol Med. 2012 Aug;30(2):351-7. doi: 10.3892/ijmm.2012.992. Epub 2012 May 9.
The aim of this study was to investigate the effects of small interference RNA (siRNA) against ribosomal protein L22 (RPL22) on ET-1-induced proliferation of human pulmonary arterial smooth muscle cells (HPASMCs). HPASMCs were transfected with siRNA against RPL22, incubated in smooth muscle cell culture medium (SMCM) and ET-1. RPL22 gene expression and protein levels of HPASMCs were measured by real-time PCR and western blotting, respectively. PCNA, CCK-8 immunohistochemistry and flow cytometry analysis were used to evaluate HPASMC proliferation. Cyclin D1 (CCND1) expression was also assayed. Following transfection with siRNA against RPL22, RPL22 expression was significantly inhibited in the control and ET-1 groups. HPASMC proliferation was significantly suppressed by transfection with siRNA against RPL22. Moreover, CCND1 was also downregulated by inhibiting RPL22 expression. In conclusion, these data suggest that inhibition of RPL22 expression can suppress HPASMC proliferation and CCND1 expression. The effects of siRNA against RPL22 on HPASMC proliferation are considered to be mediated through inhibition of CCND1 expression.
本研究旨在探讨针对核糖体蛋白 L22(RPL22)的小干扰 RNA(siRNA)对内皮素-1(ET-1)诱导的人肺动脉平滑肌细胞(HPASMC)增殖的影响。将 siRNA 转染入 HPASMC 中,在平滑肌细胞培养基(SMCM)和 ET-1 中孵育。通过实时 PCR 和 Western blot 分别测量 HPASMC 中 RPL22 基因表达和蛋白水平。通过 PCNA、CCK-8 免疫组化和流式细胞术分析评估 HPASMC 增殖。还检测了细胞周期蛋白 D1(CCND1)的表达。转染针对 RPL22 的 siRNA 后,对照和 ET-1 组中 RPL22 的表达明显受到抑制。转染针对 RPL22 的 siRNA 可显著抑制 HPASMC 增殖。此外,抑制 RPL22 表达也下调了 CCND1。综上所述,这些数据表明抑制 RPL22 表达可抑制 HPASMC 增殖和 CCND1 表达。认为 siRNA 对 HPASMC 增殖的影响是通过抑制 CCND1 表达介导的。