微小RNA145对人脐静脉血管内皮细胞增殖迁移的影响  

Effects of microRNA145 on proliferation and migration of human umbilical vein endothelial cells

作　　者：范良[1] 殷铁军[1] 李岽健[1] 张荣阁[2] 周荣[1] 周斯[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院综合医疗科,武汉430030 [2]河南省安阳市人民医院普内科

出　　处：《中华实验外科杂志》2013年第2期262-264,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81172151）;湖北省自然科学基金资助项目（2011CDBl98）

摘　　要：目的观察微小RNAl45（miRl45）对人脐静脉血管内皮细胞（HUVECs）增殖和迁移能力的影响。方法转染miRl45模拟物和阴性对照序列进入人脐静脉内皮细胞（HUVECs）细胞，采用细胞计数试剂盒一8（CCK一8）检测转染后HUVECs细胞的增殖能力，Transwell小室检测转染后HUVECs细胞的迁移能力。用实时定量聚合酶链反应（Real—timePCR）检测miRl45候选靶基因去整合素一金属蛋白酶17（ADAMl7）的mRNA表达水平，Westernblot检测ADAMl7蛋白的表达水平。结果转染48h后，转染miRl45模拟物组的吸光度（A）值（0．34±0．01）明显低于阴性对照组4值（0．44±0．0l，P〈0．01）；转染72h后，转染miRl45模拟物组的A值（0．60±0．09）也显著低于阴性对照组A值（0．86±0．05，P〈0．01）。转染miRl45组的细胞迁移穿过Transwell小室膜的细胞数[（135±6）个]明显少于阴性对照组穿过小室膜的细胞数[（205±30）个，P〈0．05]。转染miRl45模拟物组的ADAMl7的mRNA和蛋白水平均呈现下调，Real—timePCR的相对值为0．34±0．15（P〈0．01）；Westernblot灰度值为0．65±0．21（P〈0．05）。结论miRl45能够显著抑制HUVECs的增殖和迁移能力，且miRl45可能通过ADAMl7实现调控血管内皮细胞增殖和迁移功能。Objective To investigate the effect of microRNA145 （miR145） on the proliferation and migration of human umbilical vein endothelial ceils （HUVECs）. Methods MiR145 mimic and nega- tive control fragments were transfected into HUVECs via LipofectamineTM RNAiMAX. Cell counting kit-8 was used to study the proliferation of HUVECs. Transwe11 assay was used to measure the migration of HUVECs. The expression of miR145＇ s potential target gene a disintegrin and metalloproteinase 17 （ADAM17） was detected by using real-time polymerase chain reaction （Real-time PCR） and Western blotting in HUVECs. Results 4-8 h after transfection, the A values in the group transfected with miR145 mimic （0. 34 ＋0. 01 ） were significantly lower than those in negative control group （0. 44 ± 0. 01, P 〈 0. 01 ）. 72 h after transfeetion, the A values in the group transfected with miR145 mimic （0. 60 ± 0. 09） were also significantly lower than in negative control group （0. 86 ±0. 05, P 〈0. 01 ）. The number of mi- gratory cells in miR145 mimic-transfected group （135 ± 6） was significantly less than in the negative con- trol group （205±30, P 〈 0. 05）. The expression of ADAM17 mRNA and protein was downregulated in miR145 mimie-transfected group with the relative values being 0. 34 ±0. 15 （ P 〈 0.01 ） and 0. 65 ＋ 0. 21 （P 〈 0. 05 ）, respectively. Conclusion MiR145 can effectively inhibit proliferation and migration of HUVECs probably by regulating the expression of ADAM17.

关 键 词：微小RNA145 脐静脉血管内皮细胞 增殖 迁移 肿瘤血管生成 

分 类 号：R54[医药卫生—心血管疾病]