ORC1在大鼠血管平滑肌细胞增殖过程中的表达

Expression of ORC1 during proliferation of vascular smooth muscle cells in rats

目的检测不同增殖状态大鼠血管平滑肌细胞(vascular smooth muscle cells, VSMCs)中起始识别复合物1(origin recognition complex 1, ORC1)的表达.方法采用组织块贴壁法培养大鼠胸主动脉原代培养VSMCs;提取细胞总RNA,应用逆转录聚合酶链反应(RT-PCR)技术测定不同增殖状态VSMCs中ORC1 mRNA表达;采用免疫荧光、激光共聚焦显微镜检测不同增殖状态VSMCs中ORC1 蛋白表达.结果经光镜、免疫荧光鉴定证实分离培养的细胞为VSMCs;处于静止状态的VSMCs无ORC1 mRNA表达,血清刺激6 h后ORC1 mRNA表达量明显增加,12～24 h达到高峰,继后逐渐减少;处于静止期的VSMCs无ORC1蛋白表达,处于增殖状态的VSMCs中有大量ORC1蛋白表达.结论 ORC1可能在VSMCs增殖过程中起重要作用.

Objective To explore origin recognition complex 1 （ORC1） expression in the rat vascular muscle cells at different phases of proliferation. Methods Vascular smooth muscle cells （VSMCs） of thoracic aorta of rats in primary culture were obtained by the adherence method of tissue culture. Total RNA of VSMCs was extracted. The expression of ORC1 mRNA of VSMCs at different phases of proliferation was determined by reverse transcription polymerase chain reaction （RT-PCR） and the expression of ORC1 protein by immunocytochemistry and laser confocal microscopy. Results Cultured VSMCs were confirmed by light microscope and immunocytochemistry. The expression of ORC1 mRNA in the quiescence stage of VSMCs was not found. After VSMCs were stimulated with serum, the level of ORClmRNA had an obvious increase at 6 h, peaked at 12 to 24 h and decreased in the following 24 h. The expression of ORC1 protein was also not found in the quiescence stage of VSMCs, but the level of ORC1 protein during proliferation of VSMCs was significantly increased. Conclusion ORC1 may have an important role during the process of VSMCs proliferation in rats.