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超声辐照纳米聚合物体外控释DNA效果的研究

Ultrasolation of PLGA Polymeric Nanoparticles Released DNA

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【作者】 张海李莹李富荣许琴

【Author】 Zhang Hai,Li Ying,Li Furong,et al. Department of Ultrasonography,Shenzhen People s Hospital,the Second Clinical College of Jinan University,Shenzhen,Guangdong 518020 China

【机构】 深圳市人民医院暨南大学第二临床医学院超声科暨南大学第二临床医学院放射科暨南大学第二临床医学院中心实验室

【摘要】 目的利用自制携抗体结合前列腺癌细胞,探讨超声辐照PLGA纳米粒(PLGA-NPs)体外控释脱氧核糖核酸(DNA),靶向抑癌作用。方法通过溶剂蒸干法制备携有前列腺癌雄激素受体抗体的PLGA纳米粒(NPs-PLGA-GFP-AR),包裹有绿色荧光蛋白(GFP)作为标志;加入前列腺癌PC4-2细胞培养2h后,以脉冲波(pulsed wave,PW)、连续波(continuous wave,CW)超声条件进行体外辐照对比,共聚焦荧光显微镜下观测细胞转染效果。结果 NPs摄入未增加,声辐照后绿色荧光蛋白表达急剧明显增加;无超声辐照的癌细胞中5d后始见绿色荧光,超声辐照3d即可见绿色荧光;196 h后肿瘤细胞凋亡。PW1、2、3(Duty Factor,DF为10%、20%、30%,0.05~0.15 W/cm~2))组与无超声比较转染率差异显著(P<0.05),10%、20%、30%组间转染率无显著差异(P>0.05);连续波超声辐照CW1、2、3(DF为10、20、30%,声强为0.1~0.25 W/cm~2)与无超声组对照组差异显著(P<0.05),但癌细胞脱壁、死亡率高。结论超声辐照对NPs有体外促降解及靶向控释DNA作用。

【Abstract】 Objective To target DNA by using the antibody with androgen receptors,and to inhibit prostate cancer cells to grow.To study ultrasound irradiation PLGA(Polylactic/poly glycolic acid,)nano polymers(NPs)in vitro for the release of deoxyribonucleic acid(DNA)role.Methods We got PLGA nano polymer through the solvent method. With prostate cancer androgen receptors antibodies specific nanoparticles(NPs-PLGA-GFP-AR),package green fluorescent protein(GFP)as a symbol of;use PW(Pulsed Wave),CW(Continuous Wave)ultrasound,and different conditions for ultrasound irradiation in vitro,quantitative monitoring the NPs degradation and release of DNA,and cells transfection under Laser Scanning Confocal Microscope.Results We got the nano-particles join the cell culture of human prostate cancer C4-2 cells(PC4-2)two hours,then take ultrasound irradiation,nano-particles intake did not increase, but green fluorescent protein expression sharply increased obviously.without ultrasound exposure,GFP expression appeared on day 5 after addition of the nanoshells into cultured.In contrast,applying ultrasound exposure led to early expression of GFP protein on day 3 after addition of the nanoshells into cultured cells.With PWKDuty factor, DF is 10%)of ultrasonic irradiation;After 196 hours,the tumor cell apoptosis.No ultrasonic irradiation of control group and PW1(DF is 10%,sound intensity is 0.05 W/cm~2)PW2(DF is 20%,sound intensity is 0.10 W/cm~2),PW3 (DF is 30%,sound intensity is 0.15 W/cm~2);had significant difference(P<0.05).Between DF(10%,20%,30%) groups,transfection rate had no significant difference(P>0.05).CW groups with DF(10%,20%,30%)and the group with no ultrasound comparison transfection rate had significant difference(P<0.05),but the rates of death of cell were high.Conclusions Ultrasonic irradiation of NPs,has promote in vitro degradation and targeted cells controlled release DNA effect,therefore,is a promising non-invasive treatment system.

【基金】 广东省科技计划项目(No.2010B031100014)
  • 【文献出处】 中国超声医学杂志 ,Chinese Journal of Ultrasound in Medicine , 编辑部邮箱 ,2012年09期
  • 【分类号】R445.1
  • 【被引频次】3
  • 【下载频次】38
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