LHRH-PE40与癌细胞表面LHRH受体结合特性的检测

Binding Capacity of LHRH-PE40 to LHRH Receptor on Surface of Cancer Cells

目的检测LHRH-PE40融合蛋白是否保留了LHRH与癌细胞表面其相应受体特异性结合的性质。方法在Hela细胞培养液中,分别加入LHRH和LHRHPE40,经结晶紫染色法检测A值。另取宫颈癌组织切片,分别加LHRH-PE40和LHRH后,再加兔抗PE40血清和羊抗兔IgG异硫氰酸荧光素,在荧光显微镜下观察荧光强度。结果随着LHRH与LHRHPE40的摩尔数之比由100/1、250/1增加到500/1,细胞毒性的效果逐渐减弱。当LHRH/LHRH-PE40达到500/1时,肿瘤细胞表面的LHRH受体饱和;LHRH/LHRHPE40达750/1时,其A值与500/1时的A值接近。宫颈癌组织加入LHRH竞争后,荧光强度较仅加LHRHPE40组明显减弱。结论重组毒素LHRH-PE40仍保留了与癌细胞LHRH受体结合的特性。

Objective To explore the binding capacity of LHRH-PE40 to the corresponding receptor on the surface of cancer cells. Methods Add LHRH and LHRH-PE40 into Hela cell culture, stain with crystal violet and determine the A value. Add LHRH and LHRH-PE40 onto the section of cervical carcinoma tissue, then add rabbit anti-PE40 serum and fluoresein isothiocyanate-labeled goat anti-rabbit IgG and observe the fluorescence intensity under fluorescent microscope. Results Along with the increasing ratio of molar number of LHRH to LHRH-PE40, the cytotoxicity was attenuated. When the ratio reached 500/1, the receptors on surface of cancer cells were saturated. When the ratio was 750/1 ,the A value of cell culture was close to that when the ratio was 500/1. The fluorescence intensity of cervical carcinoma tissue added with both LHRH and LHRH-PE40 was significandy lower than that added with LHRH-PE40 alone. Condusion LHRH-PE40 showed binding capacity to LHRH receptor.