Silencing pyruvate kinase M2 sensitizes human prostate cancer PC3 cells to gambogic acid-induced apoptosis.
- Author:
Lei LÜ
1
;
Liang WANG
;
Guo-Song JIANG
;
Chuan-Hua ZHANG
;
Fu-Qing ZENG
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; drug effects; Carrier Proteins; genetics; metabolism; Cell Line, Tumor; Humans; Male; Membrane Proteins; genetics; metabolism; Prostatic Neoplasms; genetics; metabolism; pathology; RNA Interference; RNA, Small Interfering; Thyroid Hormones; genetics; metabolism; Xanthones; pharmacology
- From: National Journal of Andrology 2013;19(2):102-106
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effect of silencing pyruvate kinase M2 (PKM2) on gambogic acid (GA)-induced apoptosis of human prostate cancer PC3 cells.
METHODSThree specific PKM2 siRNAs and one negative control siRNA (si-NC) were transfected into PC3 cells. The silencing effect of PKM2 siRNAs was determined by real-time fluorescence quantitative PCR (qRT-PCR) and Western blot, and the effects of PKM2 siRNA on the vitality and apoptosis of GA-stimulated PC3 cells detected by MTT and AO/EB double staining, respectively. The mRNA and protein levels of c-myc and cyclin D1 were analyzed by qRT-PCR and Western blot, respectively.
RESULTSAll the 3 PKM2 siRNAs effectively reduced the mRNA and protein expressions of PKM2, and PKM2 siRNA-1 exhibited the strongest silencing effect. At 24 h after transfection, the expression levels of PKM2 mRNA and protein were reduced by 70% and 85%, respectively (P < 0.05). Twenty-four hours of treatment with GA (0.5 micromol/L) following transfection with PKM2 siRNA-1 inhibited the vitality of the PC3 cells by 68%, increased their apoptosis, and significantly down-regulated the mRNA and protein levels of c-myc (50% and 35%) and cyclin D1 (60% and 20%) (P < 0.05).
CONCLUSIONInhibition of PKM2 sensitized PC3 cells to GA-induced apoptosis, suggesting that PKM2 may be a potential therapeutic target for sensitizing human prostate cancer to GA.
