Zhang Lu, Kuang Chunyan, Yang Tianhe, Wu Qiang, Zhang Yang
Medical Examination Center, People's Hospital of Guizhou Province, Guiyang 550003, China.
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Zhonghua Xin Xue Guan Bing Za Zhi. 2014 Nov;42(11):951-6.
To explore the effect of Schlafen 1 (Slfn1) on the migration of endothelial progenitor cells (EPCs).
Rat bone marrow derived EPCs were isolated and cultured. Ad-Slfn1, ShRNA-Slfn1, ShRNA-control and Ad-control were transfected into EPCs respectively. The mRNA expression of Slfn1 and Cyclin D1 was examined by reverse transcriptase-PCR, and their protein expression was detected by Western blot. The migration of EPCs was examined by a modified Boyden chamber assay.EPCs cell cycle was determined using flow cytometry analysis.
Forty-eight hours after ShRNA-Slfn1 transfection, the mRNA and protein expression of Slfn1 in EPCs was significantly down-regulated compared to ShRNA-control EPCs (P < 0.05). Transfection of Ad-Slfn1 reversed these changes.Overexpression of Slfn1 reduced the migration capacity of EPCs while the silencing of Slfn1 by shRNA-Slfn1 increased the migration capacity of EPCs.In addition, cell cycle was arrested at G1 phase in Slfn1 overexpression group while transfection of shRNA-Slfn1 reversed these responses.Interestingly, the mRNA and protein expression of Cyclin D1 was significantly up-regulated after shRNA-Slfn1 transfection compared to ShRNA-control group (all P < 0.05), but overexpression of Slfn1 reversed these results, suggesting Cyclin D1 was involved in regulating EPCs cell cycle via Slfn1 signaling.
Slfn1 could reduce the migration capacity of EPCs via Cyclin D1 pathway.
探讨 Schlafen 1（Slfn1）对内皮祖细胞（EPCs）迁移的影响。
分离并培养大鼠骨髓来源的 EPCs。分别将 Ad-Slfn1、ShRNA-Slfn1、ShRNA-对照和 Ad-对照转染至 EPCs 中。通过逆转录聚合酶链反应检测 Slfn1 和细胞周期蛋白 D1（Cyclin D1）的 mRNA 表达，并用蛋白质免疫印迹法检测其蛋白表达。采用改良的 Boyden 小室法检测 EPCs 的迁移情况。使用流式细胞术分析 EPCs 细胞周期。
转染 ShRNA-Slfn1 48 小时后，与 ShRNA-对照 EPCs 相比，EPCs 中 Slfn1 的 mRNA 和蛋白表达显著下调（P < 0.05）。转染 Ad-Slfn1 可逆转这些变化。Slfn1 的过表达降低了 EPCs 的迁移能力，而 shRNA-Slfn1 沉默 Slfn1 则增加了 EPCs 的迁移能力。此外，Slfn1 过表达组细胞周期停滞在 G1 期，而转染 shRNA-Slfn1 可逆转这些反应。有趣的是，与 ShRNA-对照组相比，shRNA-Slfn1 转染后 Cyclin D1 的 mRNA 和蛋白表达显著上调（均 P < 0.05），但 Slfn1 的过表达逆转了这些结果，提示 Cyclin D1 通过 Slfn1 信号通路参与调节 EPCs 细胞周期。
Slfn1 可通过细胞周期蛋白 D1 途径降低 EPCs 的迁移能力。
