Effect of shRNA-mediated survivin gene silencing on apoptosis and proliferation of leukemia cell line.
- Author:
Cong-min GU
1
;
You-kai ZHU
;
Hong-yang WU
;
Meng ZHANG
;
Bing LIAO
;
Han-liang LIN
Author Information
- Publication Type:Journal Article
- MeSH: Apoptosis; drug effects; Cell Proliferation; drug effects; Gene Expression; Gene Silencing; Humans; Inhibitor of Apoptosis Proteins; Jurkat Cells; Microtubule-Associated Proteins; biosynthesis; genetics; Neoplasm Proteins; biosynthesis; genetics; RNA Interference; RNA, Messenger; biosynthesis; RNA, Small Interfering; pharmacology
- From: Chinese Journal of Hematology 2006;27(6):394-397
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo transfect a short hairpin RNA (shRNA) against survivin gene into human T lymphoblastic leukemia cell line Jurkat, and to explore the effects on apoptosis and proliferation of transfected cells.
METHODSThe survivin-shRNA expression vector were constructed and transfected into Jurkat cells. Expression of survivin mRNA and protein were assessed by RT-PCR and Western blot analysis respectively. Apoptosis index of transfected Jurkat cells was quantified by flow cytometry. The potential of cell proliferation was described by cell growth curves.
RESULTSIn survivin-shRNA transfected Jurkat cells, survivin mRNA levels were significantly reduced by 66.67% ( transient transfection) and 60.69% ( stable transfection) respectively, compared with that in control-shRNA treated group and PBS treated group (P < 0.05); and the levels of survivin protein were significantly reduced by 63.41% (transient transfection) and 60.18% (stable transfection), compared with that in the two control groups (P < 0.05). Apoptosis index was significantly increased during both transient and stable transfection, respectively [(22. 41 +/- 2.83)% and (20.73 +/- 2.56)% (P < 0.05)]. Survivin-shRNA also inhibited the proliferation of Jurkat cells.
CONCLUSIONSVector-based survivin-shRNA can effectively reduce the expression of survivin gene, induce apoptosis
