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. Copyright 2000 Wiley-Liss, Inc.
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 humanprostatic 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. Copyright 2000 Wiley-Liss, Inc.