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目的:研究重组免疫毒素LHRH -PE40与Hela细胞表面LHRH受体结合的特异性,并观察由此产生的细胞毒性作用。方法:ELISA方法检测LHRH- PE40与Hela细胞表面LHRH受体结合的高度特异性、饱合性及其与时间的关系;LHRH- PE40由LHRH受体介导对Hela细胞产生细胞毒作用进行光、电镜观察。结果:LHRH -PE40与Hela细胞表面受体的结合与正常鸡胚成纤维细胞的结合相比有显著意义(P<0. 01);LHRH- PE40与LHRH竞争LHRHR的OD值和TSH相比有显著意义(P<0. 01);光镜观察,LHRH- PE40对Hela细胞作用明显而对正常鸡胚成纤维细胞几乎没有作用;电镜观察,LHRH -PE40使Hela细胞表面出现凋亡征象。结论:与其他正常组织的细胞相比, Hela细胞膜表面LHRH受体分布呈过表达状; LHRH -PE40与细胞表面LHRH受体的结合具有高度特异性、饱和性; LHRH -PE40对细胞的作用由LHRHR介导,并且只对LHRHR阳性细胞产生毒性作用。
Objective: To investigate the specificity of LHRH-PE40, a recombinant immunotoxin, to the LHRH receptor on the surface of Hela cells and to observe the cytotoxicity. Methods: ELISA method was used to detect the specificity, saturation and LHRH-PE40 binding to LHRH receptor on HeLa cells. LHRH-PE40 mediated LHRH-mediated cytotoxicity on Hela cells, Electron microscopy. RESULTS: The binding of LHRH-PE40 to Hela cell surface receptors was significantly higher than that of normal chick embryo fibroblasts (P <0.01). LHRH-PE40 competed with LHRH for the OD value of LHRHR compared with TSH (P <0.01). Observing with light microscope, LHRH-PE40 had obvious effect on Hela cells and had almost no effect on normal chicken embryo fibroblasts. The electron microscope showed LHRH-PE40 showed signs of apoptosis on Hela cells. Conclusion: Compared with other normal cells, the distribution of LHRH receptor on Hela cell membrane surface is over-expressed. The binding of LHRH-PE40 to LHRH receptor on cell surface is highly specific and saturated. The effect of LHRH-PE40 on cells LHRHR mediated and only toxic to LHRHR positive cells.