Abstract
Aim:
To investigate whether myosin light chain kinase (MLCK) contributed to the high proliferative ability of breast cancer cells.
Methods:
Soft agar colony formation on the MCF-7 and LM-MCF-7 cell lines was determined. The cell cycles of MCF-7 and LM-MCF-7 were detected using flow cytometry analysis. Western blot analysis was performed to detect the expression levels of p-ERK1/2, total-ERK1/2, p-p38, total p38, p-JNK, total-JNK, survivin, Bcl-2, p-MLC, caspase-9, cleaved caspase-9, and MLCK. After treatment with adriamycin (ADR), ML-7 and SB203580, apoptosis was examined using flow cytometry analysis and Annexin V-FITC fluorescence microscopy.
Results:
The breast cancer LM-MCF-7 cell line with high metastasis potential (a metastitic sub-clone of MCF-7) had higher anti-apoptosis ability relative to MCF-7 cells in response to adriamycin treatment (apoptosis rate: 6.76% vs 28.58%, P<0.05). Moreover, the expression level of MLCK was upregulated and the level of phosphorylated p38 (p-p38) was decreased in LM-MCF-7 cells. Flow cytometry analysis showed that ML-7, selective inhibitor of MLCK, could induce apoptosis of the LM-MCF-7 cells, in which the level of p-p38 was increased. Meanwhile, the expression levels of Bcl-2 and survivin were downregulated, while the caspase-9 was upregulated suggesting that the cells were undergone apoptosis. Flow cytometry analysis showed that SB203580, an inhibitor of p38, abolished ML-7-induced apoptosis, which resulted in the upregualtion of Bcl-2 and survivin, and downregulation of caspase-9, suggesting that Bcl-2, survivin and caspase-9 are downstream effectors of p38.
Conclusion:
MLCK is responsible for high proliferative ability of breast cancer cells through anti-apoptosis, in which p38 pathway was involved.
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Abbreviations
- MLCK:
-
myosin light chain kinase
- p-MLC:
-
phosphorylated myosin light chain
- MAPK:
-
mitogen-activated protein kinase
- ERK:
-
extracellular signal-regulated kinase
- p-p38:
-
phosphorylated p38
- ADR:
-
adriamycin
- SCID:
-
severe combined immunodeficiency
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Acknowledgements
This work was supported from National Basic Research Program of China (973 Program, No 2007CB914804, No 2007CB914802, No 2009CB521702) and National Natural Scientific Foundation of China (No 30770826).
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Cui, Wj., Liu, Y., Zhou, Xl. et al. Myosin light chain kinase is responsible for high proliferative ability of breast cancer cells via anti-apoptosis involving p38 pathway. Acta Pharmacol Sin 31, 725–732 (2010). https://doi.org/10.1038/aps.2010.56
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DOI: https://doi.org/10.1038/aps.2010.56
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