CAJ | 학술논문

目的考察小干扰RNA(siRNA)载体促黄体激素释放激素-MPG△NLS(LHRH-MPGNLS)负载针对细胞周期素依赖性激酶2(CDK2)的siRNA所形成纳米粒的粒径和Zeta电位,观察不同溶剂对纳米粒粒径的影响,研究其对肝癌细胞HepG2的抑制效果.方法将LHRH-MPG△NLS与CDK2-siRNA按氨磷比(N/P)为10/1、20/1、40/1混合,通过自组装形成纳米粒,用动态光散射仪检测纳米粒的粒径和Zeta电位,考察焦碳酸二乙酯(DEPC)处理水、质量分数为10％葡萄糖溶液和生理盐水对纳米粒粒径的影响,用CCK8试剂盒测试纳米粒对HepG2细胞生长的抑制作用.结果 LHRH-MPG△NLS/CDK2-siRNA纳米粒的平均粒径均在200 nm以下,N/P为10/1、20/1、40/1时的Zeta电位分别为(70±5)、(120±5)、(130±5) mV.生理盐水中纳米粒粒径明显增大,说明强电解质对纳米粒粒径产生较大影响.纳米粒终浓度为200 nmol/L时,N/P为10/1的纳米粒对HepG2细胞的生长具有明显抑制作用.结论 LHRH-MPG△NLS/CDK2-siRNA纳米粒的平均粒径在200 nm以下,容易被细胞摄取,Zeta电位显示纳米粒可以在水溶液中稳定存在,但强电解质会影响纳米粒的稳定性,使其粒径增大.纳米粒终浓度为200 nmol/L时,N/P为10/1的纳米粒对HepG2细胞生长的抑制效果显著.
목적 고찰소간우RNA(siRNA)재체촉황체격소석방격소-MPG△NLS(LHRH-MPGNLS)부재침대세포주기소의뢰성격매2(CDK2)적siRNA소형성납미립적립경화Zeta전위,관찰불동용제대납미립립경적영향,연구기대간암세포HepG2적억제효과.방법 장LHRH-MPG△NLS여CDK2-siRNA안안린비(N/P)위10/1、20/1、40/1혼합,통과자조장형성납미립,용동태광산사의검측납미립적립경화Zeta전위,고찰초탄산이을지(DEPC)처리수、질량분수위10％포도당용액화생리염수대납미립립경적영향,용CCK8시제합측시납미립대HepG2세포생장적억제작용.결과 LHRH-MPG△NLS/CDK2-siRNA납미립적평균립경균재200 nm이하,N/P위10/1、20/1、40/1시적Zeta전위분별위(70±5)、(120±5)、(130±5) mV.생리염수중납미립립경명현증대,설명강전해질대납미립립경산생교대영향.납미립종농도위200 nmol/L시,N/P위10/1적납미립대HepG2세포적생장구유명현억제작용.결론 LHRH-MPG△NLS/CDK2-siRNA납미립적평균립경재200 nm이하,용역피세포섭취,Zeta전위현시납미립가이재수용액중은정존재,단강전해질회영향납미립적은정성,사기립경증대.납미립종농도위200 nmol/L시,N/P위10/1적납미립대HepG2세포생장적억제효과현저.
Objective To detect the size distribution and Zeta potential of LHRH-MPG△NLS/CDK-siRNA nanoparticles,to observe the effect of different solvents on the nanoparticle size,and to investigate the inhibitory effect of nanoparticles on HepG2 cell growth.Methods LHRH-MPG △NLS and CDK2-siRNA were mixed by continuous stirring to form nanoparticles at different N/P ratios (10/1,20/1 and 40/1).The size distribution and Zeta potential of LHRH-MPG△NLS/CDK2-siRNA nanoparticles were detected by dynamic light scattering,and the stability of the nanoparticles in normal saline,10％ glucose and pure water was discussed.Finally,the inhibitory effect of the nanoparticles on HepG2 cells was determined by CCK8 kit.Results The mean size of the nanoparticles was within 200 nm,and the Zeta potentials were (70±5) mV (N/P=10/1),(120±5) mV (N/P=20/1) and (130±5) mV (N/P=40/1),respectively.The size of the nanoparticles in normal saline was significantly increased,which demonstrated that strong electrolytes had a great impact on the nanoparticles size.When nanoparticle concentration was 200 nmol/L,LHRH-MPG△NLS/CDK2-siRNA nanoparticles (N/P=10/1) showed significantly inhibitory effect on HepG2 cell growth.Conclusions The mean size of the LHRH-MPG△NLS/CDK2-siRNA nanoparticles was within 200 nm,which was ideal for cellular uptake.The Zeta potential of nanoparticles revealed that nanoparticles could be stable in aqueous solution,while strong electrolytes would affect nanoparticle size.When nanoparticle concentration was 200 nmol/L,LHRH-MPG△NLS/CDK2-siRNA nanoparticles (N/P=10/1) showed significantly inhibitory effect on HepG2 cell growth.