@article{APS6924,
author = {Yun-wei Ou and Zi-tong Zhao and Chuan-yue Wu and Bai-nan Xu and Yong-mei Song and Qi-min Zhan},
title = {Mig-2 attenuates cisplatin-induced apoptosis of human glioma cells in vitro through AKT/JNK and AKT/p38 signaling pathways},
journal = {Acta Pharmacologica Sinica},
volume = {35},
number = {9},
year = {2016},
keywords = {},
abstract = {Yun-wei OU1, 2, Zi-tong ZHAO2, Chuan-yue WU3, Bai-nan XU1, *, Yong-mei SONG2, *, Qi-min ZHAN2
1Department of Neurosurgery, Chinese PLA General Hospital, Beijing 100853, China; 2State Key Laboratory of Molecular Oncology, Cancer Institute and Cancer Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100021, China; 3Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15261, USA
Aim: Mig-2 (also known as Kindlin-2 and FERMT2) is an important regulator of integrin activation and cell-extracellular matrix adhesion, and involved in carcinogenesis and tumor progression. The aim of this study was to investigate the role of mig-2 in cisplatin-induced apoptosis of human glioma cells in vitro.
Methods: The expression of mig-2 was modulated in human glioma H4, HS 683 and U-87 MG cells by transfection with a plasmid carrying mig-2 or mig-2 siRNA. Cisplatin-induced apoptosis was detected using Annexin V/PI staining and flow cytometry, as well as MTS analyses. The expression of apoptosis-related or signaling proteins was examined using Western blotting analysis. H4 cells were transfected with plasmids carrying mig-2 mutants to determine the functional domain of mig-2.
Results: In the 3 glioma cell lines tested, overexpression of mig-2 significantly attenuated cisplatin-induced apoptosis, whereas knock-down of mig-2 potentiated the apoptosis. The mechanisms of action of mig-2 were further addressed in H4 cells: overexpression of mig-2 markedly reduced cleaved caspase-9, caspase-8, caspase-3 and PARP, as well as p-JNK and p-p38, and increased p-AKT in cisplatin-treated H4 cells, whereas mig-2 siRNA reversely changed these apoptosis-related and signaling proteins. Furthermore, pretreatment with JNK inhibitor SP600125 and p38 inhibitor SB203580, or with AKT inhibitor LY294002 abolished the effects of mig-2 on cisplaxtin-induced apoptosis. In H4 cells, GFP-mig-2 F3 plasmid that contained only the F3 subdomain showed the same efficiency in attenuating cisplatin-induced apoptosis, as the mig-2 wild-type vector did, whereas GFP-mig-2 (1-541) plasmid that lacked the F3 subdomain was inactive.
Conclusion: Mig-2 significantly attenuates the antitumor action of cisplatin against human glioma cells in vitro through AKT/JNK and AKT/p38 signaling pathways. The F3 subdomain of mig-2 is necessary and sufficient for this effect.
Keywords: glioma; mig-2; cisplatin; apoptosis; caspases; AKT; JNK; p38; siRNA
This work is supported by the 973 National Key Fundamental Research Program of China (2012CB967003 and 2011CB910801) and the National Natural Science Foundation of China (81071633).
* To whom correspondence should be addressed.
E-mail symlh2006@163.com (Yong-mei SONG); xbn301sjwk@126.com (Bai-nan XU)
Received 2014-03-30 Accepted 2014-05-26},
issn = {1745-7254}, url = {http://www.chinaphar.com/article/view/6924}
}