中华实验外科杂志
中華實驗外科雜誌
중화실험외과잡지
CHINESE JOURNAL OF EXPERIMENTAL SURGERY
2013年
2期
262-264
,共3页
范良%殷铁军%李岽健%张荣阁%周荣%周斯
範良%慇鐵軍%李崠健%張榮閣%週榮%週斯
범량%은철군%리동건%장영각%주영%주사
微小RNA145%脐静脉血管内皮细胞%增殖%迁移%肿瘤血管生成
微小RNA145%臍靜脈血管內皮細胞%增殖%遷移%腫瘤血管生成
미소RNA145%제정맥혈관내피세포%증식%천이%종류혈관생성
microRNA145%Umbilical vein endothelial cells%Proliferation%Migration%Tumor angiogenesis
目的 观察微小RNA145 (miR145)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miR145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-8(CCK-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-time PCR)检测miR145候选靶基因去整合素-金属蛋白酶17(ADAM17)的mRNA表达水平,Westem blot检测ADAM17蛋白的表达水平.结果 转染48 h后,转染miR145模拟物组的吸光度(A)值(0.34±0.01)明显低于阴性对照组A值(0.44 ±0.01,P＜0.01)；转染72 h后,转染miR145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miR145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miR145模拟物组的ADAM17的mRNA和蛋白水平均呈现下调,Real-time PCR的相对值为0.34 ±0.15(P＜0.01)；Western blot灰度值为0.65±0.21(P ＜0.05).结论 miR145能够显著抑制HUVECs的增殖和迁移能力,且miR145可能通过ADAM17实现调控血管内皮细胞增殖和迁移功能.
目的 觀察微小RNA145 (miR145)對人臍靜脈血管內皮細胞(HUVECs)增殖和遷移能力的影響.方法 轉染miR145模擬物和陰性對照序列進入人臍靜脈內皮細胞(HUVECs)細胞,採用細胞計數試劑盒-8(CCK-8)檢測轉染後HUVECs細胞的增殖能力,Transwell小室檢測轉染後HUVECs細胞的遷移能力.用實時定量聚閤酶鏈反應(Real-time PCR)檢測miR145候選靶基因去整閤素-金屬蛋白酶17(ADAM17)的mRNA錶達水平,Westem blot檢測ADAM17蛋白的錶達水平.結果 轉染48 h後,轉染miR145模擬物組的吸光度(A)值(0.34±0.01)明顯低于陰性對照組A值(0.44 ±0.01,P＜0.01)；轉染72 h後,轉染miR145模擬物組的A值(0.60±0.09)也顯著低于陰性對照組A值(0.86±0.05,P＜0.01).轉染miR145組的細胞遷移穿過Transwell小室膜的細胞數[(135±6)箇]明顯少于陰性對照組穿過小室膜的細胞數[(205±30)箇,P＜0.05].轉染miR145模擬物組的ADAM17的mRNA和蛋白水平均呈現下調,Real-time PCR的相對值為0.34 ±0.15(P＜0.01)；Western blot灰度值為0.65±0.21(P ＜0.05).結論 miR145能夠顯著抑製HUVECs的增殖和遷移能力,且miR145可能通過ADAM17實現調控血管內皮細胞增殖和遷移功能.
목적 관찰미소RNA145 (miR145)대인제정맥혈관내피세포(HUVECs)증식화천이능력적영향.방법 전염miR145모의물화음성대조서렬진입인제정맥내피세포(HUVECs)세포,채용세포계수시제합-8(CCK-8)검측전염후HUVECs세포적증식능력,Transwell소실검측전염후HUVECs세포적천이능력.용실시정량취합매련반응(Real-time PCR)검측miR145후선파기인거정합소-금속단백매17(ADAM17)적mRNA표체수평,Westem blot검측ADAM17단백적표체수평.결과 전염48 h후,전염miR145모의물조적흡광도(A)치(0.34±0.01)명현저우음성대조조A치(0.44 ±0.01,P＜0.01)；전염72 h후,전염miR145모의물조적A치(0.60±0.09)야현저저우음성대조조A치(0.86±0.05,P＜0.01).전염miR145조적세포천이천과Transwell소실막적세포수[(135±6)개]명현소우음성대조조천과소실막적세포수[(205±30)개,P＜0.05].전염miR145모의물조적ADAM17적mRNA화단백수평균정현하조,Real-time PCR적상대치위0.34 ±0.15(P＜0.01)；Western blot회도치위0.65±0.21(P ＜0.05).결론 miR145능구현저억제HUVECs적증식화천이능력,차miR145가능통과ADAM17실현조공혈관내피세포증식화천이공능.
Objective To investigate the effect of microRNA145 (miR145) on the proliferation and migration of human umbilical vein endothelial cells (HUVECs).Methods MiR145 mimic and negative control fragments were transfected into HUVECs via LipofectamineTM RNAiMAX.Gell counting kit-8 was used to study the proliferation of HUVECs.Transwell 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 miR145mimic (0.34±0.01) were significantly lower than those in negative control group (0.44 ± 0.01,P ＜0.01).72 h after transfection,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 migratory cells in miR145 mimic-transfected group (135 ±6) was significantly less than in the negative control group (205 ± 30,P ＜ 0.05).The expression of ADAM17 mRNA and protein was downregulated in miR145 mimic-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.
