Down-regulated PTTG1 expression promotes the senescence of human prostate cancer LNCaP-AI.
- Author:
Yang-Yang WEI
1
;
Xiao-Ming SONG
2
;
Zhao-Hui XIONG
1
;
Ke-Quan LU
3
;
Lu ZHENG
3
;
Xi-Liang CAO
4
Author Information
1. Department of Urology, Anhui Armed Police Corps Hospital, Hefei, Anhui 230061, China.
2. Department of Urology, The 71st Group Army Hospital of the PLA, Xuzhou, Jiangsu 221004, China.
3. Center for Tumor Treatment, The 71st Group Army Hospital of the PLA, Xuzhou, Jiangsu 221004, China.
4. Department of Urology, Xuzhou First People's Hospital, Xuzhou, Jiangsu 221004, China.
- Publication Type:Journal Article
- Keywords:
LNCaP-AI cell;
castration-resistant prostate cancer;
cellular senescence;
pituitary tumor-transforming gene-1;
prostate cancer
- From:National Journal of Andrology
2019;25(3):216-222
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the effect of the down-regulated expression of pituitary tumor-transforming gene 1 (PTTG1) on the senescence of human castration-resistant prostate cancer LNCaP-AI cells.
METHODS:Human castration-resistant prostate cancer LNCaP-AI cells were induced in vitro and transfected with siRNA targeting PTTG1 (the siRNA-PTTG1 group), the reagent lip3000 only (the mock group) or siRNA negative control vector (the NC group). All the cells were cultured in fetal bovine serum (FBS) or charcoal-stripped bovine serum (CSS) and counted with the cell counting chamber. The senescence characteristics of the transfected LNCaP-AI cells were examined by senescence-associated β-galactosidase (SA-β-Gal) staining, and the expressions of the senescence-related β-galactosidase-1-like proteins (Glb1), the cyclin-dependent kinase inhibitors p-21CIP1 and p-27Kip1, and the chromatin-regulating heterochromatin protein 1γ (HP1γ) were detected by Western blot.
RESULTS:The expression of PTTG1 in the human prostate cancer LNCaP-AI cells was significantly reduced in the siRNA-PTTG1 group compared with those in the mock and NC groups (0.21 ± 0.01 vs 0.56 ± 0.02 and 0.61 ± 0.02, P < 0.05). Culture with FBS markedly increased while that with CSS decreased the number of LNCaP-AI cells transfected with siRNA, but both FBS and CSS enhanced the proliferation of the LNCaP-AI cells in the mock and NC groups. SA-β-Gal staining revealed that reducing the expression of PTTG1 induced a remarkably higher positive rate of the LNCaP-AI cells in the siRNA-PTTG1 than in the mock and NC groups (［63.5 ± 2.35］% vs ［11.3 ± 1.24］% and ［12.4 ± 1.15］%, P < 0.05). The siRNA-PTTG1 group, in comparison with the mock and NC groups, showed a significantly down-regulated expression of PTTG1 (0.21 ± 0.01 vs 0.56 ± 0.02 and 0.61 ± 0.02, P < 0.05), but up-regulated expressions of p-21CIP1 (0.32 ± 0.03 vs 0.20 ± 0.02 and 0.21 ± 0.03, P < 0.05), p-27Kip1 (0.38 ± 0.02 vs 0.20 ± 0.03 and 0.22 ± 0.01, P < 0.05), Glb1 (0.24 ± 0.01 vs 0.13 ± 0.01 and 0.15 ± 0.01, P < 0.05), and HP1γ (0.41 ± 0.01 vs 0.26 ± 0.01 and 0.27 ± 0.02, P < 0.05) in the LNCaP-AI cells.
CONCLUSIONS:Down-regulated expression of PTTG1 induces senescence of human castration-resistant prostate cancer LNCaP-AI cells.
