The binding characteristics of a cyclic nonapeptide, c(CGRRAGGSC), in LNCaP human prostate cancer cells

Gu,Chen; Liu,Lu; He,Yujie; Jiang,Jianwei; Yang,Zexuan; Wu,Qinghua

doi:10.3892/ol.2012.769

The binding characteristics of a cyclic nonapeptide, c(CGRRAGGSC), in LNCaP human prostate cancer cells

Authors:
- Chen Gu
- Lu Liu
- Yujie He
- Jianwei Jiang
- Zexuan Yang
- Qinghua Wu
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Affiliations: Department of Radiology, The Third Affiliated Hospital of Nantong University, Wuxi 214042, P.R. China , Nuclear Medicine Technology Institute, School of Medicine, Southeast University, Dingjiaqiao, Nanjing 210009, P.R. China , Department of Radiology, The Fourth Hospital of Wuxi, Wuxi 214062, P.R. China, Department of Radiology, The Third Affiliated Hospital of Nantong University, Wuxi 214042, P.R. China , Nuclear Medicine Technology Institute, School of Medicine, Southeast University, Dingjiaqiao, Nanjing 210009, P.R. China
Published online on: June 21, 2012 https://doi.org/10.3892/ol.2012.769
Pages: 443-449

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Abstract

Previous studies have demonstrated that interleukin-11 (IL-11) and the IL-11 receptor (IL-11R) are associated with the regulation of tumor progression and may play a significant role in bone metastases. The nonapeptide structure c(CGRRAGGSC) is a phage display-selected IL-11 mimic, which binds to IL-11R. The aim of this study is to investigate the binding characteristics of a cyclic nonapeptide c(CGRRAGGSC) in LNCaP human prostate cancer cells. To investigate its binding and uptake effects, c(CGRRAGGSC) was labeled with a fluorescent dye, LSS670. The binding location of LSS670 cyclic nonapeptide in LNCaP cells was investigated by fluorescence microscopy. Flow cytometry was used to detect the fluorescence of LSS670-c(CGRRAGGSC) in LNCaP cells. The binding of LSS670-c(CGRRAGGSC) in LNCaP cells was inhibited by unlabeled cyclic nonapeptide, depending on the varying density of c(CGRRAGGSC) and different time points. The molecular probe bound to the LNCaP cell membrane and cytoplasm through fluorescence tracing. In the saturation experiments performed in vitro, the Kd value was 3.2±0.02 nM and the Bmax value was 754±34 fmol/mg.pro. The 50% inhibiting concentration (IC50) was 6.31±0.12 nmol/l and the Ki value was 2.11±0.14 nmol/l in competitive inhibition experiments. Our results suggest that c(CGRRAGGSC) is able to specifically bind to LNCaP cells through a receptor-mediated pathway.

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