Metastatic Human Prostate Cancer Research Articles

Three-dimensional type I collagen gel system for the study of osteoblastic metastases produced by metastatic prostate cancer.

We developed a three-dimensional type I collagen gel cell culture system that allows coculturing of human MG-63 osteoblast-like cells and various human cancer cells. Inoculation of human PC-3 metastatic prostate cancer cells into this type I collagen gel containing human MG-63 osteoblast-like cells produced an osteoblastic-like reaction that presented as an increased number of MG-63 cells and increased density of type I collagen around MG-63 cells adjacent to inoculated PC-3 cells by microscope analysis. Under identical experimental conditions, inoculation of cell-free medium, human KLE endometrial adenocarcinoma cells, and Calu-1 lung cancer cells did not produce this blastic-like reaction. In situ hybridization documented the uniform expression of insulin-like growth factor I (IGF-I) and of urokinase-type plasminogen activator (uPA) mRNA in MG-63 and PC-3 cells separately cultured in this substrata. The uniform expression of uPA was also documented by immunocytochemistry using a monoclonal and a polyclonal antihuman uPA antibody. The relative expression of uPA was higher in PC-3 cells than in MG-63, KLE, and Calu-1 cancer cells. We conclude that this novel cell culture system may become a useful model to study the pathophysiology of the osteoblastic reaction in vitro.

Frequency and characterization of p53 mutations in primary and metastatic human prostate cancer.

We have studied the frequency of mutations in the p53 gene in human prostate cancer. The investigated material consisted of 20 primary-tumor tissue specimens, obtained by transurethral resection and tissue specimens of 15 lymph-node metastases, obtained at total prostatectomy. The applied methods encompassed immunohistochemistry on frozen sections, using the monoclonal antibody PAb 1801, and single-strand conformation polymorphism (SSCP) analysis, after amplification of single exon sequences by PCR, on exons 5 to 8 of the p53 gene. The mutations, leading to aberrantly migrating bands in the PCR-SSCP analysis, were identified by direct sequencing of the PCR product. Immunohistochemical and PCR-SSCP analysis were completely confirmative. In the primary tumors, mutations were found in 10% of the specimens (codons 232 and 273), and in lymph-node metastases in 15% of the specimens (codons 248 and 273). In one case (codon 273), the same mutation was found both in the primary tumor and in the lymph-node metastasis. Our results show that p53 mutations are infrequent in both primary and metastatic prostate tumors. In addition, they indicate that there is no strict correlation between p53 mutation and tumor metastasis.

Amplification, rearrangement, and elevated expression of c-myc in the human prostatic carcinoma cell line LNCaP.

We have investigated the structure and expression of the c-myc proto-oncogene in DNA isolated from the immortal cell line LNCaP. This cell line was derived from a lymph node metastasis of human prostate cancer. Msp I digest of LNCaP DNA when hybridized to a human c-myc probe showed a 1.45 kb band of intensity about two-fold greater than that observed in normal lymphocytes. In addition, the LNCaP cells contain rearranged and amplified c-myc structures which are not present in normal lymphocytes. Quantitation of these bands by scanning densitometry is consistent with an approximately 10-fold amplification of c-myc. To determine whether this amplification was accompanied by increased expression, RNA was isolated from these cells and compared to RNA isolated from a control cell line in which c-myc was not amplified. Northern blot analysis showed that the RNA transcripts from LNCaP cells were approximately 50-fold higher. Although androgens modulate the cell growth of LNCaP, there was no change in the level of c-myc RNA transcripts in serum-free medium in the presence or absence of androgens. Further investigation to determine whether altered structure, amplification, and overexpression of c-myc constitute a common characteristic of metastatic human prostate cancer might prove profitable in understanding this disease.