Prostate-specific antigen (PSA) promoter-driven androgen-inducible expression of sodium iodide symporter in prostate cancer cell lines.
Currently, no curative therapy for metastatic prostate cancer exists. Causing prostate cancer cells to express functionally active sodium iodide symporter (NIS) would enable those cells to concentrate iodide from plasma and might offer the ability to treat prostate cancer with radioiodine. Therefore, the aim of our study was to achieve tissue-specific expression of full-length human NIS (hNIS) cDNA in the androgen-sensitive human prostatic adenocarcinoma cell line LNCaP and in subcell lines C4, C4-2, and C4-2b in vitro. For this purpose, an expression vector was generated in which full-length hNIS cDNA coupled to the prostate-specific antigen (PSA) promoter has been ligated into the pEGFP-1 vector (NIS/PSA-pEGFP-1). The PSA promoter is responsible for androgen-dependent expression of PSA in benign and malignant prostate cells and was therefore used to mediate androgen-dependent prostate-specific expression of NIS. In addition, two control vectors were designed, which consist of the pEGFP-1 vector containing the PSA promoter without NIS cDNA (PSA-pEGFP-1) and NIS cDNA without the PSA promoter (NIS-pEGFP-1). Prostate cancer cells were transiently transfected with each of the above-described expression vectors, incubated with or without androgen (mibolerone) for 48 h, and monitored for iodide uptake activity. In addition, stably transfected LNCaP cell lines were established for each vector. Prostate cells transfected with NIS/PSA-pEGFP-1 showed perchlorate-sensitive, androgen-dependent iodide uptake in a range comparable to that observed in control cell lines transfected with hNIS cDNA. Perchlorate-sensitive iodide uptake was not observed in cells transfected with NIS/PSA-pEGFP-1 and treated without androgen or in cells transfected with the control vectors. In addition, prostate cancer cell lines without PSA expression (PC-3 and DU-145) did not show iodide uptake activity when transfected with NIS/PSA-pEGFP-1. Western blotting of LNCaP and C4-2b cell membranes transfected with NIS/PSA-pEGFP-1 using a monoclonal antibody that recognizes the COOH-terminus of hNIS revealed a band with a molecular weight of 90,000 that was not detected in androgen-deprived cells or in cells transfected with the control vectors, as well as a minor band at Mr 150,000 in transiently transfected LNCaP cell membranes. In conclusion, tissue-specific androgen-dependent iodide uptake activity has been induced in prostate cancer cells by PSA promoter-directed NIS expression. This study represents an initial step toward therapy of prostate cancer with radioiodine.
Spitzweg
C
C
Department of Endocrinology, Mayo Clinic, Rochester, Minnesota 55905, USA.
Zhang
S
S
Bergert
E R
ER
Castro
M R
MR
McIver
B
B
Heufelder
A E
AE
Tindall
D J
DJ
Young
C Y
CY
Morris
J C
JC
D000230:Adenocarcinoma / Q000378:metabolism / Q000473:pathology*; D000728:Androgens; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000235:genetics; D002462:Cell Membrane / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D007454:Iodides / Q000493:pharmacokinetics*; D008297:Male; D008565:Membrane Proteins / Q000096:biosynthesis* / Q000235:genetics; D009277:Nandrolone / Q000031:analogs & derivatives* / Q000494:pharmacology; D009376:Neoplasms, Hormone-Dependent / Q000378:metabolism / Q000473:pathology*; D009928:Organ Specificity; D017430:Prostate-Specific Antigen / Q000235:genetics*; D011471:Prostatic Neoplasms / Q000378:metabolism / Q000473:pathology*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics; D027981:Symporters; D014162:Transfection; D014407:Tumor Cells, Cultured / Q000187:drug effects; D000099209:Sodium-Iodide Symporters
D000728:Androgens; D002352:Carrier Proteins; D018076:DNA, Complementary; D007454:Iodides; D008565:Membrane Proteins; D009277:Nandrolone; D017430:Prostate-Specific Antigen; D011993:Recombinant Fusion Proteins; D027981:Symporters; D000099209:Sodium-Iodide Symporters; C100075:mibolerone