Neurotensin receptor expression in prostate cancer cell line and growth effect of NT at physiological concentrations

The Prostate
L SeethalakshmiRobert E Carraway

Abstract

Neurotensin (NT), a neuroendocrine peptide, exerts trophic effects in vivo and stimulates growth of some tumor cells in vitro. Androgen-sensitive prostate cells derived from lymph node carcinoma of the prostate (LNCaP) secrete NT and exhibit growth responses to NT. This study examines NT secretion, NT receptor and NT-growth responses in androgen-independent prostatic carcinoma (PC3) cells derived from prostate adenocarcinoma metastatic to bone. Binding of 125I-NT to PC3 membranes was studied by filtration. NT was measured by RIA. Reverse transcriptase polymerase chain reaction (RT-PCR) was used for NT and NT receptor mRNA. Growth was measured as 3H-thymidine incorporation into DNA. Scatchard analyses gave two binding components (Kd1 = 40 pM and Kd2 = 300 pM) in equal amounts (15-30 x 10(3) sites/cell). The bioactive region of NT was essential and the specific, non-peptide NT antagonist, SR48692, inhibited (IC50 = 3 nM). GTP analogs, sodium ion and SH-directed alkylating agents also inhibited. Glutaraldehyde crosslinking labeled two substances (M(r) of 23 and 46 kDa). RT-PCR indicated robust expression of authentic NT receptor but little for NT precursor. NT was stable in PC3 cultures but it was not found in cells or conditioned...Continue Reading

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