ClC-3 siRNA inhibits regulatory volume decrease in nasopharyngeal carcinoma cells.
- Author:
Dong YE
1
;
Hai-feng ZHANG
;
Lin-yan ZHU
;
Li-wei WANG
;
Li-xin CHEN
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line, Tumor; Cell Size; Chloride Channels; genetics; Humans; Nasopharyngeal Neoplasms; genetics; pathology; RNA Interference; RNA, Small Interfering; genetics; Transfection
- From: Journal of Southern Medical University 2011;31(2):216-220
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the role of ClC-3 chloride channels in regulatory volume decrease (RVD) of nasopharyngeal carcinoma (NPC) CNE-2Z cells.?
METHODSClC-3 siRNA was transfected into CNE-2Z cells in the presence of the transfection reagent HiPerFect Reagent(TM). The transfection efficiency of ClC-3 siRNA was detected by flow cytometry. The expression of ClC-3 protein was detected by Western blotting, and the changes of cell volume in 160 mOsmol/L hypotonic solution were determined by image analysis.
RESULTSThe transfection efficiency of ClC-3 siRNA was (63.8∓3.8)% (n=3, P<0.01), and compared with the control group, ClC-3 siRNA transfection resulted in a reduction of ClC-3 expression by (60.9∓4.0)% (n=3, P<0.01). The hypotonic challege (160 mOsmol/L) caused cell swelling and induced RVD. In the control group, hypotonic solution bath for 35 min resulted in a RVD of (42.6∓2.8)% (n=20), which was significantly decreased to (10.5∓4.8)% (n=16) in ClC-3 siRNA-transfected cells, demonstrating a reduction of RVD capacity by 75.4% (P<0.01).?
CONCLUSIONThe capacity of RVD is significantly reduced in CNE-2Z cells by ClC-3 chloride channel protein knock-down via ClC-3 siRNA transfection, indicating an important role of ClC-3 chloride channels in the RVD of CNE-2Z cells.
