Inhibitory effect of siRNA targeting ADAM17 on the proliferation of prostate cancer PC-3 cells.
- Author:
Feng LIN
1
;
Ping LIN
;
Xin LIU
;
Dong LI
;
Zi-Jun LIU
;
Hai-Feng ZOU
;
Ying JIANG
;
Xue-Fei ZHAO
;
Jin-Liang FENG
;
Xiao-Guang YU
Author Information
- Publication Type:Journal Article
- MeSH: ADAM Proteins; genetics; metabolism; ADAM17 Protein; Cell Line, Tumor; Cell Proliferation; Cyclin D1; metabolism; Cyclin-Dependent Kinase Inhibitor p21; metabolism; Down-Regulation; Humans; Male; Prostatic Neoplasms; genetics; metabolism; pathology; RNA Interference; RNA, Messenger; genetics; RNA, Small Interfering; genetics; Signal Transduction; Transfection
- From: National Journal of Andrology 2012;18(8):687-691
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo study the effect of siRNA targeting ADAM17 (ADAM17-siRNA) on the proliferation of prostate cancer PC-3 cells.
METHODSAfter transfecting PC-3 cells with ADAM17-siRNA 1 and ADAM17-siRNA 2, we detected the expressions of ADAM17 mRNA and protein by RT- PCR and Western blotting, respectively. We measured the changes in the proliferation and DNA synthesis of PC-3 cells by MTT and bromodeoxyuridine (BrdU) incorporation assay, examined the cell cycle profile by flow cytometry, and determined the expressions of the genes associated with PC-3 cell proliferation by Western blotting.
RESULTSBoth ADAM17-siRNA 1 and 2 effectively reduced the expressions of ADAM17 mRNA and protein in the PC-3 cells. Knockdown of ADAM17 with the two siRNAs significantly inhibited cell proliferation as compared with the control group (0.43 +/- 0.57 and 0.44 +/- 0.64 vs 0.80 +/- 0.51, P < 0.05) and down-regulated DNA synthesis (0.48 +/- 0.43 and 0.54 +/- 0.59 vs 0.79 +/- 0.72, P < 0.05). The cell cycle profile showed that the cell population of the G1 phase was markedly higher in both the ADAM17-siRNA groups than in the control ([61.83 +/- 2.41]% and [59.78 +/- 1.92]% vs [41.38 +/- 1.53]%, P < 0.05), but that of the S phase remarkably lower in the former two than in the latter ([23.64 +/- 2.56]% and [25.24 +/- 1.86]% vs [33.51 +/- 1.47]%, P < 0.05), with a concomitant decrease in the expression of the cell cycle protein cyclin D1 and increase in the cyclin-dependent kinase inhibitor p21.
CONCLUSIONADAM17-siRNA can effectively inhibit the proliferation of PC-3 cells by up-regulating cyclin D1 and down-regulating p21 protein, and ADAM17 has a potential value in the gene therapy of prostate cancer.
