CAJ | 학술논문

目的:通过对核糖体失活蛋白Gelonin进行化学修饰,利用穿膜肽和聚乙二醇(PEG)偶联来提高其到达肿瘤部位和进入肿瘤细胞的能力,使Gelonin更高效地发挥抑瘤作用. 方法:利用FPLC Superdex75分子筛预装柱纯化系统对所修饰的Gelonin进行纯化后,在不同细胞系测试细胞毒性;通过倒置荧光显微镜、流式细胞分析技术等对药物进入纤维肉瘤细胞HT1080的能力进行评价;采用小动物活体成像技术考察药物体系在荷瘤动物体内的分布情况. 结果:采用分子筛色谱纯化可以得到纯度相对较高的修饰产物,其毒性较无修饰的Gelonin强,且在HT1080细胞系作用最明显;细胞摄取结果显示,与未修饰的Gelonin相比,该药物体系具有更高的细胞摄取效率;动物成像结果表明,PEG5000修饰可以改变Gelonin在动物体内的分布情况,增加在肿瘤的药物蓄积.结论:穿膜肽和PEG5000修饰后的Gelonin有较高的肿瘤细胞摄取和杀伤能力,药物在肿瘤的蓄积量较高,从而增强了药物的抑瘤效果.
목적:통과대핵당체실활단백Gelonin진행화학수식,이용천막태화취을이순(PEG)우련래제고기도체종류부위화진입종류세포적능력,사Gelonin경고효지발휘억류작용. 방법:이용FPLC Superdex75분자사예장주순화계통대소수식적Gelonin진행순화후,재불동세포계측시세포독성;통과도치형광현미경、류식세포분석기술등대약물진입섬유육류세포HT1080적능력진행평개;채용소동물활체성상기술고찰약물체계재하류동물체내적분포정황. 결과:채용분자사색보순화가이득도순도상대교고적수식산물,기독성교무수식적Gelonin강,차재HT1080세포계작용최명현;세포섭취결과현시,여미수식적Gelonin상비,해약물체계구유경고적세포섭취효솔;동물성상결과표명,PEG5000수식가이개변Gelonin재동물체내적분포정황,증가재종류적약물축적.결론:천막태화PEG5000수식후적Gelonin유교고적종류세포섭취화살상능력,약물재종류적축적량교고,종이증강료약물적억류효과.
Objective:To improve anti-tumor effect of Gelonin, the plant-sourced RIP is modified by chemically conjugating a cell-penetrating peptide and polyethylene glycol (PEG). Methods:Purified protein was obtained after being performed on FPLC (fast protein liquid chromatography) Superdex75 column. Cytotoxicity was detected by MTT assay. The cellular uptake by HT1080 cells was studied by using inverted fluorescence microscopy and flow cytometry. In-vivo imaging technology was utilized for investigation of the in-vivo drug distribution in the HT1080 tumor-bearing mice. Results:The modified product was purified by using gel filtration chromatergraphy. Moreover, compared with native Gelonin, the cytotoxicity of modified protein was increased, especially in HT1080, presumably due to the enhanced cellular uptake. The in-vivo imaging results suggested that drug accumulation in tumor was improved by PEGylation. Conclusion:Modified Gelonin can improve cell penetration and cytotoxicity in tumor cells. PEGylation can increase tumor accumulation of the protein drug, and thereby enhance its anti-tumor effect.