IL11类似物c(CGRRAGGSC)与人前列腺癌PC-3细胞体外结合特性的研究  

Binding characteristics of interleukin 11 analogue-cyclic nonapeptide c(CGRRAGGSC) to human prostate cancer PC-3 cells:An in vitro study

作　　者：何玉洁[1] 吴清华[2,3] 顾琛[3] 姜建威[3] 

机构地区：[1]无锡市第四人民医院影像中心,江苏无锡214002 [2]东南大学医学院核医学技术研究所,江苏南京210009 [3]南通大学第三附属医院影像科,江苏无锡214002

出　　处：《中华男科学杂志》2011年第4期330-335,共6页National Journal of Andrology

摘　　要：目的:探讨白细胞介素11(IL11)类似物环九肽c(CGRRAGGSC)与人前列腺癌PC-3细胞的体外结合特性。方法:采用荧光染料LSS670标记c(CGRRAGGSC)合成LSS670-c(CGRRAGGSC),流式细胞仪测定其与PC-3细胞体外结合的荧光强度,计算其竞争抑制试验中的半数抑制量(IC50)和平衡抑制常数(K i),荧光显微镜观察LSS670-c(CGRRAGGSC)在PC-3细胞的定位;用99mTc标记c(CGRRAGGSC)合成99mTc-DTPA-c(CGRRAGGSC)与PC-3细胞进行放射受体结合分析,Scatchard作图法计算标记物与PC-3细胞结合的平衡解离常数(Kd)和每个细胞上的最大结合位点数(Bm ax)。结果:LSS670-c(CGRRAGGSC)与人前列腺癌PC-3细胞的结合具有可饱和性,浓度与时间呈依赖性。未标记c(CGRRAGGSC)与LSS670-c(CGRRAGGSC)对PC-3受体具有竞争性抑制作用[IC50=(6.31±0.12)nmol/L,Ki=(2.11±0.14)nmol/L]。LSS670-c(CGRRAGGSC)荧光主要集中在PC-3细胞膜上,Kd值为(0.11±0.02)nmol/L,Bm ax为(230±34)fmol/mg pro。结论:c(CGRRAGGSC)符合特异性配体的标准。Objective： To investigate the binding characteristics of interleukin 11 （IL-11 ） analogue-cyclic nonapeptide c（ Cys- Gly-Arg-Arg-Ala-Gly-Gly-Ser-Cys） NH2 C30H54N16010S2, c（CGRRAGGSC） , and human prostate cancer PC-3 cells. Methods： c （CGRRAGGSC） was labeled with fluorescent dye LSS670, and the location of LSS670-cyclic nonapeptide in the PC-3 cells was investigated by fluorescent microscopy. Flow cytometry was used to detect the fluorescence intensity of the in vitro binding of LSS670-c （CGRRAGGSC） to PC-3 cells and calculate its IC50 and Ki in competitive inhibition experiments. 99Tcm-DTPA-c（CGRRAGGSC） was synthesized by the reaction of ^99mTc04- with c（CGRRAGGSC）. The binding characteristics of ^99mTc-DTPA-c（CGRRAGGSC） and IL11R in the PC-3 cells were analyzed by radioreceptor assay. Bmax and Kd were calculated in saturability and reversibility experiments. Results ： The binding of LSS670-c（CGRRAGGSC） to the PC-3 cells showed the characteristics of saturability and concentration-time dependence. Unlabeled c（CGRRAGGSC） and LSS670-c（CGRRAGGSC） exhibited a competitive inhibition on the PC-3 cells （IC50 = [ 6.31± 0.12 ] nmol/L, Ki = [ 2.11 ± 0.14 ] nmol/L）. Fluorescence was mainly distributed in the cell membrane （Kd = [0.32 ±0.02] nmol/L, Bmax = [754 ±34] fmol/mg pro）. Conclusion： e（CGRRAGGSC） could bind PC-3 cells through a receptor-mediated pathway.

关 键 词：PC-3细胞株 白细胞介素11类似物 核素 荧光染料 前列腺癌 

分 类 号：R737.25[医药卫生—肿瘤]