Binding characteristics of interleukin 11 analogue-cyclic nonapeptide c(CGRRAGGSC) to human prostate cancer PC-3 cells: an in vitro study.
- Author:
Yu-Jie HE
1
;
Qing-Hua WU
;
Chen GU
;
Jian-Wei JIANG
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line, Tumor; Humans; Interleukin-11; metabolism; Male; Peptides; metabolism; Prostatic Neoplasms; metabolism; Protein Binding
- From: National Journal of Andrology 2011;17(4):330-335
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the binding characteristics of interleukin 11 (IL-11) analogue-cyclic nonapeptide c(Cys-Gly-Arg-Arg-Ala-Gly-Gly-Ser-Cys) NH2 C30H54N16O10S2, c(CGRRAGGSC), and human prostate cancer PC-3 cells.
METHODSc(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 99mTcO4- 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.
RESULTSThe 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).
CONCLUSIONc (CGRRAGGSC) could bind PC-3 cells through a receptor-mediated pathway.
