Short hairpin RNA-targeting special AT-rich sequence-binding protein-1 induces the apoptosis of human lung cancer cell line A549

2012 
Objective: To investigate the effect of short hairpin RNA (shRNA)-targeting special AT-rich sequence-binding protein-1 (SATB1) on the apoptosis of human lung cancer cell line A549, and to explore the possible mechanism. Methods: The recombined plasmid SATB1-shRNA of shRNA-targeting SATB1 was constructed and transfected into A549 cells by LipofectAMINE 2000. The expression levels of SATB1, Bcl-2 and Bax mRNAs were examined by RT-PCR, and the expression levels of SATB1, Bcl-2, Bax, and caspase 3 proteins were examined by Western-blotting. The apoptosis rate of A549 cells was detected by flow cytometry (FCM). Results: The recombined plasmid SATB1-shRNA was successfully constructed and transfected into the A549 cells. After transfection with SATB1-shRNA, the expression levels of SATB1 and Bcl-2 mRNAs and proteins were decreased (P<0.05), whereas the expression levels of Bax mRNA and protein and the caspase 3 protein were increased (P<0.05). The apoptosis rate of A549 cells after transfection with SATB1-shRNA was higher than that of the A549 cells without transfection with SATB1-shRNA [(14.18±1.59) % vs (1.84±0.57) %, P<0.01]. Conclusion: SATB1-shRNA can significantly down-regulate the expression level of SATB1 in human lung cancer cell line A549 and induce the apoptosis. The mechanism may be related to the cascade effect induced by the down-regulation of Bcl-2 expression. DOI:10.3781/j.issn.1000-7431.2012.07.004
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