Alteration of ERalpha expression and tumor invasion by RNA interference against metastasis associated-1 gene in human breast cancer cells.
- Author:
Qing-ming JIANG
1
;
Hui ZHANG
;
Ping ZHANG
Author Information
- Publication Type:Journal Article
- MeSH: Blotting, Western; Breast Neoplasms; genetics; metabolism; pathology; Cell Line, Tumor; Cell Movement; drug effects; Cell Proliferation; drug effects; Down-Regulation; drug effects; Estrogen Receptor alpha; genetics; metabolism; Histone Deacetylases; genetics; Humans; Matrix Metalloproteinase 9; genetics; metabolism; RNA Interference; physiology; RNA, Messenger; metabolism; RNA, Small Interfering; pharmacology; Repressor Proteins; genetics; Reverse Transcriptase Polymerase Chain Reaction
- From: Chinese Journal of Pathology 2008;37(2):118-123
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the inhibitory effect on MTA1 gene by MTA1 short hairpin RNA (shRNA), downstream regulation of expression of ERa and invasion of human breast cancer cells.
METHODSRecombinant plasmid pGenesil-1/MTA1 was constructed and transfected into human breast cancer cell lines MDA-MB-231 and MCF-7. Fluorescence microscopy was used to evaluate the efficacy of transfection. The transcription expression of MTA1 and MMP-9 gene were determined by reverse transcription-polymerase chain reaction (RT-PCR), the protein expression of ERalpha and MMP-9 were determined by immunohistochemical and western blot. The invasion ability was evaluated by Boyden chamber invasive assay.
RESULTSRecombinant plasmid pGenesil-1/MTA1 was constructed and transfected into MDA-MB-231 (82.5%) and MCF-7 (78.2%) successfully. After the transfection, MTA1 mRNA was suppressed in both cell lines (80.2% and 58.7%). The expression of ERalpha protein became positive in transfected MDA-MB-231 cell, and the expression of MMP-9 mRNA were down-regulated. The invasion ability was decreased (27.2% +/- 2.1)% compared with (76.3% +/- 2.4%), (P < 0.05). In contrast, transfected MCF-7 cells failed to show significant difference in the expression of ERalpha and MMP-9, without change of the invasion ability (P > 0.05).
CONCLUSIONSRNA interference can effectively suppress the expression of MTA1 in human breast cancer cells. An induction of ERalpha protein and suppression of MMP-9 may be related to the decrease of tumor cell invasive ability. RNA interference involving MTA1 gene may provide an effective anti-cancer gene therapy.