ORC1在大鼠血管平滑肌细胞G1/S和S期的表达

目的检测大鼠血管平滑肌细胞(VASCULAR SMOOTH MUSCLE CELLS,VSMCS)G1/S、S期中起始识别复合物(ORIGINRECOGNITION COMPLEX1,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。利用胸苷双阻断法造成细胞同步,应用逆转录聚合酶链反应(RT-PCR)技术、流式细胞仪检测分别测定G1/S、S期VSMCS的ORC1MRNA及蛋白表达。结果经光镜、免疫组化鉴定证实分离培养的细胞为VSMCS。ORC1MRNA在GO期的VSMCS无明显表达、G1/S期达到高峰,到S、G2/M期明显减少。流式细胞仪检测的VSMCS ORC1蛋白质表达与MRNA变化相似。结论ORC1可能在VSMCS细胞周期调节过程中起重要作用。

Expression of ORC1 at G1/S and S phases in rat vascular smooth muscle cells

Objective To explore the expression of origin recognition complex 1 (ORC1) at G_ 1 /S and S phases in vascular smooth muscle cells (VSMCs) of rats. Methods VSMCs of thoracic aorta of rats were primarily cultured by the method of tissue adherence. The cell synchrony was obtained by the method of double-thymidine block. Total RNA of VSMCs was extracted, and the ORC1 mRNA expression at G_ 1 /S and S phases was determined by RT-PCR. The ORC1 protein expression was measured by flow cytometry. Results Cultured VSMCs were confirmed by light microscope and immunocytochemistry. The ORC1 mRNA expression was not found at G_ 0 phase of VSMCs, peaked at G_ 1 /S phase, and decreased at S and G_ 2 /M phases. The change of ORC1 protein by flow cytometry was the same as ORC1 mRNA in cell cycle. Conclusion ORC1 may have an important role during the process of VSMCs cell cycle in rats.