Expression profile and functional activity of peptide transporters in prostate cancer cells.

Abstract

Peptide transporters are expressed predominantly in intestinal and renal epithelial cells. The functional expression of peptide transporters is also identified in other types of tissues, such as glia cells, macrophages, and the epithelia of the bile duct, the lungs, and the mammary glands. However, their presence and role are poorly understood in carcinomas. We explored the expression profile and functional activity of peptide transporters in the prostate cancer cell lines LNCaP, PC-3, and DU145. Quantitative real time RT-PCR (qRT-PCR) and Western blot were used to evaluate the expression profile of peptide transporter 1 (PEPT1), peptide transporter 2 (PEPT2), peptide histidine transporter 1 (PHT1), and peptide histidine transporter 2 (PHT2) in these cells. LNCaP expresses high levels of PEPT2 and PHT1, while PC-3 demonstrates strong expression of PEPT1 and PHT1. DU145 shows only weak expression of PEPT1 and PHT1. Functional activities were studied in these cell lines using radiolabeled glycylsarcosine ([(3)H]Gly-Sar) and l-histidine ([(3)H]-l-histidine). The uptake of [(3)H]Gly-Sar and [(3)H]-l-histidine was time- and pH-dependent. A kinetic study showed that the uptake of Gly-Sar and l-histidine is saturable over the tested concentration range. The binding affinity (K(m)) and the maximal velocity (V(max)) exhibited in the three cell lines were consistent with the expression profiles we observed in qRT-PCR and Western blot analysis. A competitive inhibition study revealed that peptide transporters in prostate cancer cells exhibited broad substrate specificity with a preference for hydrophobic dipeptides, such as Leu-Leu. Fluorescence microscopy study revealed that the fluorescent dipeptide probe d-Ala-Lys-AMCA (a substrate of peptide transporters) specifically accumulated in the cytoplasm of LNCaP and PC-3, but not DU145 cells. Inhibiting the peptide transporter activity by Gly-Sar suppressed the growth of LNCaP and PC-3 cells. Our study indicated that PC-3 cells can be established as a new cell culture model for PEPT1 study, and LNCaP can be used as a model for PEPT2 study. Moreover, our results suggested that peptide transporters are overexpressed in prostate cancer cells and can be adopted as a promising target for tumor-specific drug delivery.

• Authors: Wanyi Tai, Zhijin Chen, Kun Cheng
• Journal: Molecular pharmaceutics (Mol Pharm) Vol. 10 Issue 2 Pg. 477-87 (Feb 04 2013) ISSN: 1543-8392 [Electronic] United States
• PMID: 22950754 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Carrier Proteins
• Membrane Transport Proteins
• Nerve Tissue Proteins
• Peptide Transporter 1
• SLC15A3 protein, human
• SLC15A4 protein, human
• Symporters
• hydrogen-coupled oligopeptide transporter PepT2

Topics

• Blotting, Western
• Caco-2 Cells
• Carrier Proteins (genetics, metabolism)
• Cell Line, Tumor
• Cell Proliferation
• Humans
• Male
• Membrane Transport Proteins (genetics, metabolism)
• Nerve Tissue Proteins (genetics, metabolism)
• Peptide Transporter 1
• Prostatic Neoplasms (genetics, metabolism)
• Real-Time Polymerase Chain Reaction
• Symporters (genetics, metabolism)