RNA干扰沉默基质交感分子1基因后对血管平滑肌细胞周期的影响

Effect of Stromal Interaction Molecule 1 Knockdown on Cell Cycle of Vascular Smooth Muscle Cell

目的探讨RNA干扰技术沉默基质交感分子1基因后对血管平滑肌细胞周期的影响。方法原代培养血管平滑肌细胞,用构建好的大鼠基质交感分子1干扰腺病毒载体和人重组基质交感分子1腺病毒载体进行转染,Western blot检测细胞基质交感分子1、p21和pRb表达,流式细胞仪测定细胞周期。结果大鼠基质交感分子1干扰腺病毒载体转染后48h细胞基质交感分子1表达与转染对照腺病毒载体比较明显降低(P<0.05),处于G0/G1细胞比例也明显增多(P<0.05),p21表达上调,pRb表达下调(P<0.05);人重组基质交感分子1腺病毒载体共转染后48h细胞基质交感分子1表达恢复到正常水平;G0/M细胞比例、p21和pRb表达恢复到正常水平。结论基质交感分子1基因沉默上调p21表达,下调pRb表达,抑制细胞进入S期,进而参与对血管平滑肌细胞增殖的调控。

Aim To explore the effect of STIM1 knockdown on cell cycle of vascular smooth muscle cell. Methods Rat VSMC was isolated and primary cultured. Ad-si/rSTIM1 and Ad-hSTIM1 were transfected into VSMC. The protein of STIM1, p21 and pRb were measured by western blot, the cell cycle of VSMC was analyzed by flow cytometry. Results On 48 h after transfected Ad-si/rSTIM1, the expression of STIM1 protein was decreased significantly compared to that in Ad-hSTIM1-treated group （P〈0.05）. The percent of G0/G1 phase cell was significantly increased （P〈0.05）. The expression of p21 protein was increased and the expression of pRb protein was decreased （P〈0.05）. However, the cotransfection of Ad-hSTIM1 with Ad-si/rSTIM1 restored these responses. Conclusions STIM1 knockdown up-regulates the expression of p21 and down-regulates the expression of pRb, which inhibits cell to progress into S phase. Stim1 maybe have a key role in the vascular smooth muscle cell proliferation.