CAJ | 학술논문

目的构建重组靶向抗肿瘤肽(Citrostatin),研究其生物学活性.方法构建Citrostatin重组基因,经原核生物表达和纯化获得Citrostatin融合蛋白,通过内皮细胞杀伤实验、细胞毒活性实验、体外管状机构生成抑制实验等分析Citrostatin的生物学活性.结果 Citrostatin融合蛋白经表达、酶切纯化后,目的蛋白纯度达90%以上.Citrostatin 能明显抑制内皮细胞ECV304的增殖(IC_(50)=2.28μmol/L),有效杀伤肿瘤细胞1990及NCI-H640(IC_(50)分别为9.24,2.7μmol/L),并明显抑制体外管状结构的生成.结论成功构建了重组靶向抗肿瘤肽Citrostatin,细胞和血管水平研究表明该融合肽同时具有抑制肿瘤新生血管的形成和直接杀伤肿瘤的双重活性.
목적 구건중조파향항종류태(Citrostatin),연구기생물학활성.방법 구건Citrostatin중조기인,경원핵생물표체화순화획득Citrostatin융합단백,통과내피세포살상실험、세포독활성실험、체외관상궤구생성억제실험등분석Citrostatin적생물학활성.결과 Citrostatin융합단백경표체、매절순화후,목적단백순도체90%이상.Citrostatin 능명현억제내피세포ECV304적증식(IC_(50)=2.28μmol/L),유효살상종류세포1990급NCI-H640(IC_(50)분별위9.24,2.7μmol/L),병명현억제체외관상결구적생성.결론 성공구건료중조파향항종류태Citrostatin,세포화혈관수평연구표명해융합태동시구유억제종류신생혈관적형성화직접살상종류적쌍중활성.
Purpose To construct a recombined antitumor peptide and to analyze its bioactivity. Methods Constructing a recombined gene and inserting the pGEX-4T-3 vector. The recombined protein was expressed in E. coli BL21 and purified with Amylose Resin. Then, citrostatin was subjected to the following tests separately: inhibition of endothelial cell proliferation, MTT test of cytotoxicity and inhibition of endothelial cell tube formation on ECMatrix. Results Citrostatin significantly inhibited the proliferation of human endothelial cell ECV304(IC_(50) = 2.28 μmol/L) .It also significantly inhibited the proliferation of human tumor cell 1990 and NCI-H64O(IC_(50) = 9.24,2.74 μmol/L) ,and the inhibitory effect became more marked with the increase of citrostatin concentration. The inhibitory effects of citrostatin on endothelial cell tube formation was also confirmed . Conclusion An antitumor peptide, citrostatin, has been successfully constructed and purified, which showed anti-angiogenesis effect and direct cytotoxic effect on tumor cells.