CAJ | 학술논문

目的：探讨由DMRIE-C脂质体介导将CLEC2B siRNA转染入Jrukat细胞的过程中，对Jurkat细胞的影响，并分析其转染率的影响因素。方法通过DMRIE-C转染CLEC2B siRNA入Jrukat细胞，台盼蓝染色法观察细胞活性，流式细胞仪检测siRNA转染率，RT-PCR检测CLEC2B基因沉默效率。结果 DMRIE-C脂质体对细胞的损伤程度与两者比例相关，对转染体系600μL （其中200μL密度为2×106／mL淋巴细胞）进行转染时，脂质体达3μL时可损伤细胞，不利于后续试验进行；siRNA∶DMRIE-C比例为3μL∶2μL，siRNA终浓度约80 nmol／L时，转染率最高，可达30．7％，转染后CLEC2B基因抑制率可达（35±1．9）％（t＝15．9，P＜0．05）。结论 DMRIE-C脂质体可实现目的siRNA对Jurkat细胞有效转染，同时转染中脂质体与细胞比例、血清、操作方式等因素均可对转染效率产生一定的影响。
목적：탐토유DMRIE-C지질체개도장CLEC2B siRNA전염입Jrukat세포적과정중，대Jurkat세포적영향，병분석기전염솔적영향인소。방법통과DMRIE-C전염CLEC2B siRNA입Jrukat세포，태반람염색법관찰세포활성，류식세포의검측siRNA전염솔，RT-PCR검측CLEC2B기인침묵효솔。결과 DMRIE-C지질체대세포적손상정도여량자비례상관，대전염체계600μL （기중200μL밀도위2×106／mL림파세포）진행전염시，지질체체3μL시가손상세포，불리우후속시험진행；siRNA∶DMRIE-C비례위3μL∶2μL，siRNA종농도약80 nmol／L시，전염솔최고，가체30．7％，전염후CLEC2B기인억제솔가체（35±1．9）％（t＝15．9，P＜0．05）。결론 DMRIE-C지질체가실현목적siRNA대Jurkat세포유효전염，동시전염중지질체여세포비례、혈청、조작방식등인소균가대전염효솔산생일정적영향。
Objective To observe the process of transfecting CLEC 2B siRNA mediated by DMRIE-C liposome into the Jurkat cells and to analyze the factors of influencing transfection rate .Methods We transfected CLEC2B siRNA into the Jurkat cells by DMRIE-C.Trypan blue staining was used to observe the cell activity .siRNA transfection efficiency was de-tected by flow cytometry , and the CLEC2B gene silencing efficiency was detected by RT-PCR.Results The cell damage degree in the transfection process was influenced by the cell members /transfection system lipo concentration ratio , 3μl lipo in the transfection system (600 μl total liquid, 200 μl 2 ×106/ml lymphocytes) would damage cells, and go against the following experiment.The transfection efficiency could increase to 30.70%when the siRNA:DMRIE-C ratio was adjusted to 3 μl∶2 μl, and then, the CLEC2B gene expression inhibition ratio was about 35 ±1.9%(t=15.9, P<0.05).Con-clusion DMRIE-C lipo cloud successfully transfer aim siRNA to Jurkat cells , and meanwhile, lipo/lymphocyte ratio, ser-um interference and operation skills may influence the transfection efficiency .