siRNAs对原代人脐静脉内皮细胞血管生成的影响  

Effects of siRNAs on angiogenesis of primary human umbilical vein endothelial cells

作　　者：陈佳扬 黎权辉 岑柏宏 陈章浦 杨伶 庞建新[1] 付卫明[1] 季爱民[2] CHEN Jia-yang;LI Quan-hui;CEN Bo-hong;CHEN Zhang-pu;YANG Ling;PANG Jian-xin;FU Wei-ming;JI Ai-min(School of Pharmacy,Southern Medical University,Guangzhou 5J0515,Guangdong Province,China;Department of Pharmacy,The Seventh Affiliated Hospital of Southern Medical University,Foshan 528200,Guangdong Province,China;Department of Radiation Oncology,Affiliated Cancer Hospital of Guangzhou Medical University,Guangzhou 510095,Guangdong Province,China)

机构地区：[1]南方医科大学药学院,广东510515 [2]南方医科大学第七附属医院药学科,广东佛山528200 [3]广州医科大学附属肿瘤医院放疗科,广东广州510095

出　　处：《中国临床药理学杂志》2022年第9期904-907,共4页The Chinese Journal of Clinical Pharmacology

基　　金：国家自然科学基金资助项目(81773641)。

摘　　要：目的 研究siRNA单独或多重沉默原代人脐静脉内皮细胞(HUVECs)中血管内皮细胞生长因子A(VEGFA)及血管内皮细胞生长因子受体1/2/3(VEGFR1/2/3)表达对其血管生成的影响。方法 HUVECs分为空白对照组,阴性对照组与实验组。空白对照组不给予转染,阴性对照组转染100 nmol·L^(-1)siNC,实验组分为4组并分别转染100 nmol·L^(-1)siVEGFA及siVEGFR1/2/3。以反转录-聚合酶链反应(RT-PCR)检测siRNAs的沉默效率。HUVECs分为空白对照组,阴性对照组,沉默单靶点组和多靶点组,空白对照组不给予转染,阴性对照组转染100 nmol·L^(-1)siNC,沉默单靶点组分为4组并分别转染100 nmol·L^(-1)siVEGFA及siVEGFR1/2/3,沉默多靶点组分为4组并分别转染100 nmol·L^(-1)siVEGFA+siVEGFR1、siVEGFA+siVEGFR2、siVEGFA+siVEGFR3及siVEGFR1+siVEGFR2+siVEGFR3,以小管形成实验检测各组HUVECs小管形成情况。结果 siVEGFA组、siVEGFR1组、siVEGFR2组与siVEGFR3组沉默效率分别为(85.67±10.50)%,(71.67±7.02)%,(89.00±11.36)%与(90.00±3.60)%,与空白及阴性对照组相比,差异均有统计学意义(均P<0.01)。siVEGFA组血管总长度、分支管长度和血管交叉点分别为(32.67±3.79)%,(22.67±2.08)%及(25.33±6.43)%,在沉默单靶点组中抑制血管生成能力最强(均P<0.05);siVEGFA+siVEGFR3组血管总长度、分支管长度和血管交叉点分别为(28.33±5.51)%,(10.00±2.65)%,(8.33±2.31)%,siVEGFR1+siVEGFR2+siVEGFR3组分别为(27.00±4.00)%,(9.33±2.89)%,(10.00±3.60)%,在沉默多靶点组中抑制血管生成能力最强(均P<0.05)。结论 siRNAs沉默VEGF通路中单靶点的表达,可有效抑制HUVECs诱导的血管生成;同时沉默多靶点的表达,其抑制血管生成的功效更强。Objective To study the effects of single or multiple silencing the expression of vascular endothelial cell growth factor A(VEGFA) and vascular endothelial cell growth factor receptor 1 /2 /3(VEGFR1 /2 /3) on angiogenesis of human umbilical vein endothelial cells (HUVECs) by siRNA.Methods HUVECs were divided into blank control group,negative control group and experimental group.The blank control group was not treated.The negative control group was transfected with 100 nmol · L^(-1)si NC.The experimental group was transfected with 100 nmol · L^(-1)siVEGFA and siVEGFR1 /2 /3respectively.The silencing efficiency was determined by reverse transcription -polymerase chain reaction (RT -PCR) .HUVECs were divided into blank control group,negative control group,silencing single -target groups and silencing multi -target groups.The blank control group was not treated.The negative control group was transfected with 100nmol·L^(-1)si NC.The silencing single -target groups were divided into 4 groups and transfected with 100 nmol·L^(-1)si VEGFA and si VEGFR1 /2 /3 respectively.The silencing multi -target groups were divided into 4 groups and transfected with 100 nmol · L^(-1)si VEGFA + si VEGFR1, si VEGFA + si VEGFR2, si VEGFA + si VEGFR3 and si VEGFR1 + si VEGFR2 + si VEGFR3 respectively.The angiogenesis inhibition was analyzed by Image J.Results The silencing efficiency of si VEGFA and si VEGFR1 /2 /3 groups were (85.67±10.50) % ,(71.67±7.02) % ,(89.00±11.36) % and (90.00±3.60) % respectively.Compared with blank control group and negative control group,they showed statistical differences (all P<0.01) .The ratios of total length,branching length and junctions in si VEGFA group were (32.67±3.79) % ,(22.67±2.08) % ,(25.33±6.43) % and the inhibition of angiogenesis was the strongest in silencing single -target groups (all P<0.05) .The ratios of total length,branching length and junctions in si VEGFA + si VEGFR3 group were (28.33±5.51) % ,(10.00±2.65) % ,(8.33±2.31) % ,which in si VEGFR1 + si VEGFR2 + si VEGFR3 group

关 键 词：原代人脐静脉内皮细胞 血管内皮细胞生长因子A 血管内皮细胞生长因子受体 小干扰RNA 血管生成 

分 类 号：R97[医药卫生—药品]