Effects of blocking androgen receptor expression with specific hammerhead ribozyme on in vitro growth of prostate cancer cell line.
- Author:
Qiangsong TONG
1
;
Jun ZHAO
;
Zhaohui CHEN
;
Fuqing ZENG
;
Gongcheng LU
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; Cell Line, Tumor; Gene Transfer Techniques; Genetic Therapy; methods; Genetic Vectors; Humans; Immunohistochemistry; Male; Prostatic Neoplasms; therapy; RNA, Catalytic; physiology; Receptors, Androgen; analysis; genetics; Reverse Transcriptase Polymerase Chain Reaction; Transfection
- From: Chinese Medical Journal 2003;116(10):1515-1518
- CountryChina
- Language:English
-
Abstract:
OBJECTIVETo study the possibility of gene therapy for prostate cancer by blocking androgen receptor (AR) gene expression using a specific hammerhead ribozyme (RZ).
METHODSThe hammerhead ribozyme expression vector pcDNA-hAR-RZ, specific to AR mRNA, was constructed and transfected into the prostate cancer cell line LNCaP by using lipofectamine. Androgen receptor expression was measured by RT-PCR and immunohistochemical methods. Cellular proliferation activities were assayed using the tetrazolium bromide colorimetry method; cell cycle changes were observed by flow cytometry; and cell apoptosis was detected by the TdT-mediated dUTP-biotin nick end labeling method.
RESULTSOne to seven days after transfection with the ribozyme expression vector, AR mRNA expression at molecular and protein levels in LNCaP cells decreased by 32.6% - 40.7% (P < 0.05) and 21.0% - 87.64% (P < 0.05) respectively, and cell proliferation was inhibited by 18.28% - 35.34% (P < 0.05). Meanwhile, the cell cycle was arrested at the G2/M stage, and apoptotic morphological changes occurred with an apoptosis rate of 25.17% (P < 0.01).
CONCLUSIONRibozyme specific against AR mRNA is capable of inhibiting the expression AR and inducing the apoptosis in prostate cancer cells.