超顺磁性脂质体载血管内皮细胞生长因子沉默基因对骨肉瘤细胞影响  

Effects of vascular endothelial cell growth factor gene silencing in superparamagnetic liposomes on osteosarcoma cells

作　　者：黄涛 罗聪[1] 罗玉佳 陈韬 余杰 HUANG Tao;LUO Cong;LUO Yu-jia;CHEN Tao;YU Jie(Department of Orthopedics,Children’s Hospital of Chongqing Medical University,National Clinical Research Center for Child Health and Disorders,Ministry of Education Key Laboratory of Child Development and Disorders,Chongqing Engineering Research Center of Stem Cell Therapy,Chongqing 400014,China;Department of Neurosurgery,Children’s Hospital of Chongqing Medical University,National Clinical Research Center for Child Health and Disorders,Ministry of Education Key Laboratory of Child Development and Disorders,Chongqing Engineering Research Center of Stem Cell Therapy,Chongqing 400014,China)

机构地区：[1]重庆医科大学附属儿童医院,国家儿童健康与疾病临床医学研究中心,儿童发育疾病研究教育部重点实验室,重庆市干细胞治疗工程技术研究中心,骨科,重庆400014 [2]重庆医科大学附属儿童医院,国家儿童健康与疾病临床医学研究中心,儿童发育疾病研究教育部重点实验室,重庆市干细胞治疗工程技术研究中心,神经外科,重庆400014

出　　处：《中国临床药理学杂志》2022年第22期2693-2697,共5页The Chinese Journal of Clinical Pharmacology

基　　金：重庆市科技局技术创新与应用发展专项面上基金资助项目(cstc2019jscx-msxmX0121)。

摘　　要：目的探讨超顺磁性脂质体载血管内皮细胞生长因子(VEGF)沉默基因对骨肉瘤细胞影响。方法制备磁性脂质体,检测脂质体磁性和Zeta电位,透射电镜观察脂质体形态。骨肉瘤细胞143B分成空白对照组(常规培养)、阴性对照组(磁性脂质体转染siRNA control)、Lip 2000-VEGF组(Lipofectamine 2000转染siRNA VEGF)、MCLs组(磁性脂质体转染siRNA VEGF)。噻唑蓝(MTT)法检测增殖,实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测VEGF mRNA,蛋白质印迹(Western blot)法检测VEGF蛋白表达,流式细胞术测定凋亡,划痕愈合实验检测划痕愈合能力,Transwell小室检测侵袭和迁移。结果随着磁场强度的不断增加,磁性脂质体磁化强度也明显增加;磁性脂质体Zeta电位测定结果为正电位。脂质体呈圆形或椭圆形,分布均匀,分散性较好。空白对照组、阴性对照组、Lip 2000-VEGF组、MCLs组VEGF mRNA表达水平为1.00±0.01,1.05±0.03,0.43±0.02,0.24±0.01,VEGF蛋白表达水平为1.00±0.01,1.03±0.01,0.66±0.02,0.46±0.02;细胞光密度值(增殖活性)为1.97±0.12,2.01±0.21,1.51±0.12,0.96±0.10;凋亡率为(4.97±0.06)%,(6.25±0.09)%,(12.08±0.11)%,(21.15±0.12)%;划痕愈合率为(95.43±0.21)%,(86.10±0.21)%,(45.25±0.09)%,(26.69±0.20)%;细胞迁移数目为(1241.33±3.51),(1154.67±8.14),(736.33±6.43),(416.00±7.00)个;细胞侵袭数目为(986.33±5.03),(1007.33±8.02),(574.67±12.90),(248.33±7.51)个。上述指标,Lip 2000-VEGF组、MCLs组与Control比较,差异均有统计学意义(均P<0.05);MCLs组与Lip 2000-VEGF组比较,差异均有统计学意义(均P<0.05)。结论超顺磁性脂质体载VEGF沉默基因抑制骨肉瘤细胞增殖、迁移和侵袭,诱导细胞凋亡。Objective To investigate the effect of superparamagnetic liposome-loaded vascular endothelial cell growth factor(VEGF)silencing gene on osteosarcoma cells.Methods Magnetic liposomes were prepared;the magnetic properties and Zeta potential of liposomes were detected;liposome morphology was observed by transmission electron microscope.Osteosarcoma cells 143B were divided into control group(blank control),si-NC group(magnetic liposome transfected siRNA control),Lip 2000-VEGF group(Lipofectamine 2000 transfected siRNA VEGF),MCLs group(magnetic liposome transfected siRNA VEGF).Methyl thiazolyl tetrazolium(MTT)was used to detect proliferation;VEGF mRNA was detected by real-time quantitative reverse transcription polymerase chain reaction method;VEGF protein expression was detected by Western blot;apoptosis was detected by flow cytometry;scratch healing ability was detected by scratch healing assay,and invasion and migration were detected by Transwell chamber.Results The magnetization of the magnetic liposomes also increased significantly with the increase of the magnetic field intensity.The Zeta potential of the magnetic liposomes was positive.The liposomes were round or oval with uniform distribution and good dispersibility.The levels of VEGF mRNA in control,si-NC,Lip 2000-VEGF,and MCLs groups were 1.00±0.01,1.05±0.03,0.43±0.02,0.24±0.01;the VEGF protein levels were 1.00±0.01,1.03±0.01,0.66±0.02,0.46±0.02;the optical density value(proliferative activity)were 1.97±0.12,2.01±0.21,1.51±0.12,0.96±0.10;the apoptosis rate were(4.97±0.06)%,(6.25±0.09)%,(12.08±0.11)%,(21.15±0.12)%;the scratch healing rate were(95.43±0.21)%,(86.10±0.21)%,(45.25±0.09)%,(26.69±0.20)%;the number of cell migration was 1241.33±3.51,1154.67±8.14,736.33±6.43,416.00±7.00;the number of cell invasion were 986.33±5.03,1007.33±8.02,574.67±12.90,248.33±7.51.Compared Lip 2000-VEGF group and MCLs group with control group,the differences of the obove indicators were statistically significant(all P<0.05);compared MCLs group with and

关 键 词：磁性脂质体 血管内皮细胞生长因子 骨肉瘤 迁移 凋亡 

分 类 号：R28[医药卫生—中药学]