Experimental study of K562 cell apoptosis induced by siRNA / 中华血液学杂志
Chinese Journal of Hematology
;
(12): 717-719, 2004.
Article
in Chinese
| WPRIM
| ID: wpr-229912
ABSTRACT
<p><b>OBJECTIVES</b>To construct a siRNA expression vector pBCR6 that produces siRNA against bcr/abl mRNA and detect apoptosis rate of K562 cells after pBCR6 transfection.</p><p><b>METHODS</b>Template sequence for siRNA was designed, synthesized and inserted into an expression vector pSilencer1.0-U6. Restriction analysis and sequencing were performed to verify the pBCR6 vector. Then pBCR6 was transfected into K562 cells by X-tremeGene Q2. pSilencer1.0-U6 was used as the control. At different time point after transfection, apoptosis rate was determined by Tunel and Annexin V+ PI with FCM.</p><p><b>RESULT</b>pBCR6 was verified by restriction analysis and sequencing. The apoptosis rate of K562 cells markedly increased at 48 and 72 hour after transfected with pBCR6, and increased in a time-dependent manner [the apoptosis rate of transfected K562 cells was (47.80 +/- 1.63)% at 72 hrs, whereas the control group was (6.67 +/- 0.37)%, P < 0.0001] No prominent change in apoptosis rate was found in the control.</p><p><b>CONCLUSION</b>The siRNA expression vector against bcr/abl mRNA was successfully constructed. The pilot study showed that pBCR6 could effectively induce K562 cells apoptosis. siRNA may be a new tool for molecular target therapy for chronic myelogenous leukemia.</p>
Full text:
Available
Index:
WPRIM (Western Pacific)
Main subject:
Plasmids
/
RNA, Messenger
/
Base Sequence
/
Transfection
/
Fusion Proteins, bcr-abl
/
Apoptosis
/
K562 Cells
/
In Situ Nick-End Labeling
/
RNA, Small Interfering
/
Flow Cytometry
Limits:
Humans
Language:
Chinese
Journal:
Chinese Journal of Hematology
Year:
2004
Type:
Article
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