EXPRESSION OF PROSTATIC FACTORS MEASURED BY REVERSE TRANSCRIPTION-POLYMERASE CHAIN-REACTION IN HUMAN-PAPILLOMAVIRUS TYPE-18 DEOXYRIBONUCLEIC-ACID IMMORTALIZED PROSTATE CELL-LINES

Objectives. To investigate expression of the prostatic markers prostat e-specific antigen (PSA), prostate-specific membrane antigen (PSM), an d the androgen receptor (AR) after human papillomavirus (HPV) type 18 deoxyribonucleic acid (DNA) transfection and subsequent immortalizatio n of human prostate epithelial cells. Methods. Recently, two human pro state epithelial cell lines were established by HPV transformation: PZ -HPV-7, derived from normal peripheral zone (PZ) tissue, and CA-HPV-IO , derived from high Gleason grade adenocarcinoma. Expression of PSA wa s studied by the reverse transcription polymerase chain reaction (RT-P CR), because in preliminary studies using immunocytochemistry and Nort hern blotting, no PSA expression was found. PSM was analyzed by RT-PCR and nested RT-PCR. These analyses included primary human prostate cel l strains. Furthermore, androgen-supplemented methylthiazol tetrazoliu m (MTT) growth assays were performed and expression of AR was studied by immunocytochemistry. Prostate carcinoma cell lines LNCaP and PC-346 C were included as positive controls and breast carcinoma cell line MC F-7 as a negative control. Results. Both cell lines exhibited low leve ls of RNA for PSA and PSM in comparison with cell lines LNCaP and PC-3 46C. AR expression by immunocytochemistry was negative using monoclona l antibody F39.4 and polyclonal antibody SP-197. In an androgen-supple mented environment, growth rates of both HPV immortalized cell lines w ere not stimulated in contrast to LNCaP. Conclusions. RNA transcripts of PSA and PSM were detected by RT-PCR in HPV immortalized prostate ep ithelial cell lines PZ-HPV-7 and CA-HPV-10. The expression of prostate -specific markers may further validate the utility of this stepwise tr ansformation model of human prostate carcinogenesis. (C) 1998, Elsevie r Science Inc. All rights reserved.