Small interfering RNA-mediated MAPK p42 silencing induces apoptosis of HeLa cells.
- Author:
Chen HUANG
1
;
Li-ying LIU
;
Tu-sheng SONG
;
Lei NI
;
Li-ping SONG
;
Lü-sheng SI
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; physiology; Gene Silencing; physiology; HeLa Cells; Humans; Mitogen-Activated Protein Kinase 1; biosynthesis; genetics; RNA, Small Interfering; genetics; Transfection
- From: Journal of Southern Medical University 2006;26(1):11-15
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo observe the effect of small interfering RNA (siRNA)-induced MAPK p42 silencing on the survival of HeLa cells.
METHODSTwo siRNAs targeting at the MAPK p42 gene and one random siRNA were synthesized respectively by Silencer siRNA Construction Kit and transfected into HeLa cells by Lipofectamin 2000. The expression of p42(MAPK) in the transfected HeLa cells was analyzed by Western blotting and immunohistochemistry, and the morphology of cells were observed with electron microscope. TUNEL assay and Annexin V/PI staining were employed for detecting the cell apoptosis.
RESULTSThe expression of p42(MAPK) in the HeLa cells was remarkably suppressed after transfection with the two siRNAs, reduced by about 2.5 and 3.2 folds respectively in comparison with the negative control. Chromatin margination in the cell nuclei were observed in the transfected cells, and TUNEL assay and Annexin V/PI staining further confirmed the occurrence of cell apoptosis.
CONCLUSIONIn vitro MAPK p42 siRNA-1 and siRNA-2 transfection can specifically silence the gene expression and induce apoptosis of HeLa cells.
