miR-320通过下调SRY相关高迁移率族蛋白4的表达抑制人脐静脉内皮细胞内皮间质转化

miR-320 inhibits endothelial-mesenchymal transition of human umbilical vein endothelial cells by down-regulating the expression of SRY-related high mobility group protein 4

目的探讨miR-320对人脐静脉内皮细胞(HUVEC)内皮间质转化(EndMT)的影响及其调控机制。方法用miR-320 mimics或miR-320 inhibitor处理HUVEC。Western blot检测内皮细胞标记物血小板内皮细胞黏附分子1(CD31)、血管内皮钙黏蛋白(VE-Cadherin)及间质细胞标记物α平滑肌肌动蛋白(α-SMA)、波形蛋白(Vimentin)的表达;划痕实验、Transwell实验检测miR-320对HUVEC迁移的影响;罗丹明鬼笔环肽染色检测miR-320对HUVEC细胞骨架的影响;噻唑蓝法检测细胞增殖;流式细胞仪检测细胞周期;Western blot检测miR-320对SRY相关高迁移率族蛋白4(SOX4)表达的影响。结果miR-320 mimics上调HUVEC的VE-Cadherin、CD31表达,下调Vimentin、α-SMA表达;miR-320 mimics使HUVEC细胞骨架微丝变细,应力纤维减少,细胞迁移能力减弱,但增殖能力无明显改变。miR-320 inhibitor下调HUVEC的VE-Cadherin、CD31表达,上调Vimentin、α-SMA表达;miR-320 inhibitor使HUVEC微丝增粗,应力纤维增多,细胞迁移能力增强。miR-320 mimics下调SOX4的表达,miR-320 inhibitor上调SOX4的表达。结论miR-320通过下调SOX4的表达抑制HUVEC的EndMT。

Aim To investigate the effect of miR-320 on endothelial-mesenchymal transition(EndMT)of human umbilical vein endothelial cell(HUVEC)and its regulatory mechanism.Methods HUVECs were treated with miR-320 mimics or miR-320 inhibitor.Western blot was used to detect the expressions of endothelial cell markers platelet endothelial cell adhesion molecule-1(CD31)and vascular endothelial cadherin(VE-Cadherin),and the expressions of mesenchymal cell markersα-smooth muscle actin(α-SMA)and Vimentin.Scratch test and Transwell experiment were used to detect the effect of miR-320 on HUVEC migration.The effect of miR-320 on cytoskeleton of HUVEC was detected by Rhodamine phalloidin staining.Cell proliferation was detected by methyl thiazolyl tetrazolium method and cell cycle was detected by flow cytometry.Western blot was used to detect the effect of miR-320 on the expression of SRY-related high mobility group protein 4(SOX4).Results miR-320 mimics up-regulated the expressions of VE-Cadherin and CD31,and down-regulated the expressions of Vimentin andα-SMA in HUVEC.miR-320 mimics made HUVEC cytoskeleton microfilaments thinner,stress fibers decreased,cell migration ability weakened,but the proliferation ability did not change significantly.miR-320 inhibitor down-regulated the expressions of VE-Cadherin and CD31,and up-regulated the expressions of Vimentin andα-SMA in HUVEC.miR-320 inhibitor made HUVEC microfilaments thicker,stress fibers increased and cell migration ability enhanced.miR-320 mimics down-regulated SOX4 expression,and miR-320 inhibitor up-regulated SOX4 expression.Conclusion miR-320 inhibits EndMT of HUVEC by down-regulating SOX4 expression.