CAJ | 학술논문

目的：进行外源 FeIT 基因转染人白血病细胞缺乏 FeIT 表达的 eL60，研究 FeIT 基因对转染细胞生长的生物学影响。方法：构建 PEGFP - FeIT 真核表达质粒（实验组）与质粒 PEGFP - N1（对照组）。分别电穿孔法转染 eL60细胞。应用荧光显微镜、RT - PCR、Western blot 检测转染基因的转录和表达情况。四甲基偶氮唑盐（MTT）法、流式细胞术研究转染基因对 eL60细胞体外增殖、凋亡情况的影响，并与转染对照质粒的细胞株进行比较。结果：经 PCR、酶切和 DNA 测序证实 PEGFP - FeIT 真核载体构建成功。荧光显微镜下实验组 eL60细胞可见绿色荧光，转染率为40%。RT - PCR 和 Western blot 分别从 mRNA 和蛋白水平检测到 FeIT的表达。MTT 检测结果显示：转染后实验组抑制率明显升高，细胞增殖受到抑制（ P ﹤0.05）。流式细胞术结果显示：实验组细胞凋亡率明显增加（P ﹤0.05）。结论：转染 FeIT 基因对白血病细胞 eL60的生长有抑制作用，并能诱导其凋亡。
목적：진행외원 FeIT 기인전염인백혈병세포결핍 FeIT 표체적 eL60，연구 FeIT 기인대전염세포생장적생물학영향。방법：구건 PEGFP - FeIT 진핵표체질립（실험조）여질립 PEGFP - N1（대조조）。분별전천공법전염 eL60세포。응용형광현미경、RT - PCR、Western blot 검측전염기인적전록화표체정황。사갑기우담서염（MTT）법、류식세포술연구전염기인대 eL60세포체외증식、조망정황적영향，병여전염대조질립적세포주진행비교。결과：경 PCR、매절화 DNA 측서증실 PEGFP - FeIT 진핵재체구건성공。형광현미경하실험조 eL60세포가견록색형광，전염솔위40%。RT - PCR 화 Western blot 분별종 mRNA 화단백수평검측도 FeIT적표체。MTT 검측결과현시：전염후실험조억제솔명현승고，세포증식수도억제（ P ﹤0.05）。류식세포술결과현시：실험조세포조망솔명현증가（P ﹤0.05）。결론：전염 FeIT 기인대백혈병세포 eL60적생장유억제작용，병능유도기조망。
Objective:The eL60 of acute myelogenous leukemia cell lines lacked FeIT exPression,to determine the biological effects of FeIT gene exPression in eL60. Methods:We constructed a eukaryotic exPression vector PEG-FP - FeIT and transfected it into the eL60 cells by electroPoration transfect. Fluorescence microscoPe,RT - PCR and Western blot were used to examine the exPression of recombinant Plasmid at mRNA and Protein levels after transfec-tion. MTT assay and flow cytometry were used to determine the effects of FeIT exPression on the Proliferation and aP-oPtosis. Results:A correct eukaryotic exPression vector PEGFP - FeIT was identified by PCR,endoenzyme analysis and sequence of DNA. The GFP exPression was observed by fluorescence microscoPy,transfection efficiency was 40% . The FeIT mRNA and Protein could be detected within the transfected cells by RT - PCR and Western blot. Re-sults of MTT assay indicated that the exPression of FeIT significantly inhibited the Proliferation of eL60 cells in the exPerimental grouP(P ﹤ 0. 05). Flow cytometry Proved that the aPoPtosis index of FeIT - eL60 was higher than the control grouP. Conclusion:Re - exPression of FeIT gene using gene transfection technology effectively inhibits the growth and Proliferation of eL60 cell,and induces aPoPtosis of eL60 cell.