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Effect of c-FLIP-L on the sensitivity of breast cancer MDA-MB-231 cells to TRAIL-induced apoptosis
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Abstract:
Objective : To study the effect of c-FLIP-L on TRAIL-induced breast cancer apoptosis. Methods: c-FLIP-L was silenced in breast cancer MDA-MB-231 cells by siRNA. After c-FLIP-L silencing, cell proliferation was assessed by MTT assay, cell apoptosis by Annexin-V FITC/PI double staining flow cytometry, invasive potential by matrigel invasion assay, and protein and mRNA levels of c-FLIP-L, caspase-3, caspase-8, MMP-2, MMP-9 in transfected cells by Western blotting and RT-PCR respectively. Results: The sequence-specific siRNA significantly decreased c-FLIP-L mRNA (3712±3.02 vs 183.21±8.31, 174.65±10.06; P<0.05) and protein levels as compared with the control. At 72 h after treatment, c-FLIP-L siRNA and TRAIL, either in combination or each alone, significantly inhibited proliferation (\[75.51±2.01\]% vs \[33.75±1.60\]%, \[34.31±2.01\]%; P<0.01), induced apoptosis (\[76.30±4.11\]% vs \[38.95±2.14\]%, \[29.28±1.66\]%; P<0.05), decreased the invasive capacity. Enhanced casepase-3 and casepase-8 expression,and inhibited MMP-2 and MMP-9 expression in MDA-MB-231 cells. Conclusion: Inhibition of c-FLIP-L expression may increase the sensitivity of breast cancer cells to TRAIL-induced apoptosis, possibly through enhancing the expression of caspase-3, caspase-8 and inhibiting the expression of MMP-2 and MMP-9.
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Project supported by the Natural Science Foundation of Liaoning Province （No.2013022051）
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