Efficient DNA-mediated gene transfer into prostate cancer cell line LNCaP.
BACKGROUND
Prostate cancer is the most commonly diagnosed cancer and the second leading cause of cancer death in men. In the beginning stages of the disease, prostate cancer is dependent on androgens for growth. The only common and readily available cell line to study this phase of prostate cancer in vitro is LNCaP, which was originally derived from a lymph node metastatis of a human prostatic adenocarcinoma. However, DNA-mediated gene transfer, a common and key procedure in cellular and molecular studies, is very inefficient for LNCaP cells, and this limits the utility of these cells in investigations of the molecular mechanisms of prostate carcinogenesis.


METHODS
In search of a simple, reproducible, and cost-effective method for introducing DNA into LNCaP cells, we adopted and optimized two methods of transient transfection into LNCaP cells: a modified calcium phosphate (CaPO(4)) coprecipitation procedure and polyethylenimine (pEI)-mediated transfection.


RESULTS
When compared with the liposome-mediated transfection that was previously used for LNCaP cells, we find that the most efficient of these techniques is the modified CaPO(4) coprecipitation procedure. For experiments in which calcium exposure of the cells is not desirable, the pEI procedure provides a less efficient, but reproducible and cost-effective alternative.


CONCLUSIONS
These two new DNA-mediated gene transfer methods should facilitate gene expression studies in LNCaP cells and thereby aid in the study of the androgen-dependent phase of prostate cancer.
Frønsdal
K
K
Biotechnology Center of Oslo, University of Oslo, Norway.
Engedal
N
N
Saatcioglu
F
F
D004247:DNA / Q000502:physiology*; D018014:Gene Transfer Techniques; D005821:Genetic Techniques; D006801:Humans; D008297:Male; D011471:Prostatic Neoplasms / Q000235:genetics* / Q000473:pathology; D014162:Transfection / Q000379:methods; D014407:Tumor Cells, Cultured
D004247:DNA