Wang Wenhui, Li Xinming, Liu Liang, Hou Jiayin, Zhu Qian, Cong Xinpeng
Department of Cardiology, Shanghai Pudong New Area Zhoupu Hospital, Shanghai 200092, China.
Department of Cardiology, Shanghai Pudong New Area Zhoupu Hospital, Shanghai 200092, China. Email:
Zhonghua Xin Xue Guan Bing Za Zhi. 2015 Feb;43(2):157-61.
The purpose of this study is to explore the impact of stromal interaction molecule 1 (STIM1) knockdown on the proliferation and migration capacities of endothelial progenitor cells (EPCs).
The rat bone marrow derived EPCs were obtained and divided into three groups: adenovirus negative control (NSC) group, rat STIM1 adenovirus vector transfection (si/rSTIM1) group and rat and human recombinant STIM1 adenovirus transfection (si/rSTIM1+hSTIM1) group. The STIM1 expressions in each group were detected by reverse transcription PCR after transfection. The cell proliferation was tested by [(3)H] thymidine incorporation assay ((3)H-TdR). Cell cycle was analyzed by flow cytometry. The cells migration activity was detected by Boyden assay. Calcium ion concentration was detected by confocal laser scanning microscopy.
48 h after transfection, the expression level of STIM1 in si/rSTIM1 group was significantly lower than that in NSC group (0.21 ± 0.12 vs. 1.01 ± 0.01, P < 0.05), and number of EPCs at G1 phase in si/rSTIM1 group ((93.31 ± 0.24)%) was significantly higher than that in NSC group ((78.03 ± 0.34)%, P < 0.05), and EPCs' migration activity in si/rSTIM1 group (10.03 ± 0.33) was significantly lower than that in NSC group (32.11 ± 0.54, P < 0.05), and EPCs calcium ion concentration in EPCs in si/rSTIM1 group (38.03 ± 0.13) was significantly lower than that in NSC group (98.11 ± 0.34, P < 0.05), while there was no significant difference between si/rSTIM1+hSTIM1 group and NSC group on the above four indexes.
Silencing STIM1 could attenuate EPCs proliferation and migration capacities by modulating the calcium ion concentration in EPCs.
本研究旨在探讨基质相互作用分子1（STIM1）基因敲低对内皮祖细胞（EPCs）增殖和迁移能力的影响。
获取大鼠骨髓来源的EPCs并分为三组：腺病毒阴性对照组（NSC）、大鼠STIM1腺病毒载体转染组（si/rSTIM1）和大鼠及人重组STIM1腺病毒转染组（si/rSTIM1+hSTIM1）。转染后通过逆转录PCR检测各组中STIM1的表达。采用[³H]胸腺嘧啶核苷掺入法（³H-TdR）检测细胞增殖。通过流式细胞术分析细胞周期。采用Boyden试验检测细胞迁移活性。通过共聚焦激光扫描显微镜检测钙离子浓度。
转染48小时后，si/rSTIM1组中STIM1的表达水平显著低于NSC组（0.21±0.12对1.01±0.01，P<0.05），si/rSTIM1组中处于G1期的EPCs数量（(93.31±0.24)%）显著高于NSC组（(78.03±0.34)%，P<0.05），si/rSTIM1组中EPCs的迁移活性（10.03±0.33）显著低于NSC组（32.11±0.54，P<0.05），si/rSTIM1组中EPCs的钙离子浓度（38.03±0.13）显著低于NSC组（98.11±0.34，P<0.05），而si/rSTIM1+hSTIM1组和NSC组在上述四项指标上无显著差异。
沉默STIM1可通过调节EPCs中的钙离子浓度来减弱EPCs的增殖和迁移能力。
