CAJ | 학술논문

目的：新型阳离子核酸载体 LW13-2进行研究，分析其细胞毒性和基因转染效率，探讨其作为核酸载体的可能性。方法使用 MTT 比色法分析 LW13-2与 DNA 在不同比例下的细胞毒性。选用绿色荧光蛋白表达质粒 EGFP 为报告基因，以人胚胎肾 HEK293细胞为工具细胞，在荧光显微镜下观察计数，计算转染效率。结果当 LW13-2与 DNA 的质量比为（1～3）∶1时，细胞毒性最低。LW13-2与 DNA 的质量比为3∶1时，转染作用4 h，在无血清培养基中继续培养48 h，得到 LW13-2的最大转染效率为87.6％。结论通过与市售成熟转染试剂 Lipofectamine 2000的对比，新型阳离子核酸载体在大幅提高基因转染效率的同时并没有增加其细胞毒性，有望成为核酸转移的有效载体，为相关研究及基因的靶向治疗研究提供基础。
목적：신형양리자핵산재체 LW13-2진행연구，분석기세포독성화기인전염효솔，탐토기작위핵산재체적가능성。방법사용 MTT 비색법분석 LW13-2여 DNA 재불동비례하적세포독성。선용록색형광단백표체질립 EGFP 위보고기인，이인배태신 HEK293세포위공구세포，재형광현미경하관찰계수，계산전염효솔。결과당 LW13-2여 DNA 적질량비위（1～3）∶1시，세포독성최저。LW13-2여 DNA 적질량비위3∶1시，전염작용4 h，재무혈청배양기중계속배양48 h，득도 LW13-2적최대전염효솔위87.6％。결론통과여시수성숙전염시제 Lipofectamine 2000적대비，신형양리자핵산재체재대폭제고기인전염효솔적동시병몰유증가기세포독성，유망성위핵산전이적유효재체，위상관연구급기인적파향치료연구제공기출。
Objective To deal with cell transfection and cytotoxicity for LW13-2,a new cationic liposome which was prepared by our laboratory .Methods Cytotoxicity of LW13-2 was evaluated by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT)colori-metric assay.Furthermore,we chose pEGFP that could express the enhanced green fluorescent protein as a reporter to observe the transfection efficiency directly.Then,LW13-2 complexes were transfected into cells as HEK 293 and compared with Lipofectamine 2000(Invitrogen)which was widely used in cell transfection .The efficiency was measured by FACS.Results When the ratio was 3∶1 that LW13-2∶DNA ,we got the lowest cytotoxicity.And after the liposome without serum acted 4 hours,we incubated the cells for 48 hours,and got the highest gene delivery of 87.6%.Conclusion The prepared new cationic liposome can improve the transfection efficiency sharply and keep the low cytotoxicity.It shows considerable potential as an efficient nonviral carrier for gene delivery and provides a basis for gene therapy.