TY - JOUR
T1 - Cucurbitacin e induces caspase-dependent apoptosis and protective autophagy mediated by ROS in lung cancer cells
AU - Ma, Guixin
AU - Luo, Weiwei
AU - Lu, Jinjian
AU - Ma, Dik Lung
AU - Leung, Chung Hang
AU - Wang, Yitao
AU - Chen, Xiuping
N1 - Funding Information:
This study was supported by the Science and Technology Development Fund, Macao S.A.R (FDCT) ( 039/2014/A1 ) and the Research Fund of University of Macau [ MYRG118(Y2-L4)-ICMS13-CXP ].
PY - 2016/6/25
Y1 - 2016/6/25
N2 - Cucurbitacin E (CuE) is a triterpenoid with potent anticancer activities while the underlying mechanisms remain elusive. In the present study, the anticancer effects of CuE on 95D lung cancer cells were investigated. CuE decreased cell viability, inhibited colony formation, and increased reactive oxygen species (ROS) in a concentration-dependent manner, which were reversed by N-acetyl-l-cysteine (NAC). CuE induced apoptosis as determined by JC-1 staining, expression of Bcl-2 family proteins, cleavage of caspases, and TUNEL staining. NAC and Ac-DEVD-CHO partially reversed CuE-induced cleavage of caspase-3, caspase-7, and PARP. Furthermore, CuE caused accumulation of autophagic vacuoles and concentration- and time-dependent expression of LC3II protein. Autophagy inhibitors chloroquine and bafilomycin A1 enhanced CuE-induced LC3II expression and cell death. CuE-triggered protein expression of p-AKT, p-mTOR, Beclin-1, and p-ULK1 was partially reversed by NAC pretreatment. In addition, CuE treatment damaged F-actin without affecting β-tubulin as confirmed by immunofluorescence. In conclusion, CuE induced ROS-dependent apoptosis through Bcl-2 family and caspases in 95D lung cancer cells. Furthermore, CuE induced protective autophagy mediated by ROS through AKT/mTOR pathway. This study provides novel roles of ROS in the anticancer effect of CuE.
AB - Cucurbitacin E (CuE) is a triterpenoid with potent anticancer activities while the underlying mechanisms remain elusive. In the present study, the anticancer effects of CuE on 95D lung cancer cells were investigated. CuE decreased cell viability, inhibited colony formation, and increased reactive oxygen species (ROS) in a concentration-dependent manner, which were reversed by N-acetyl-l-cysteine (NAC). CuE induced apoptosis as determined by JC-1 staining, expression of Bcl-2 family proteins, cleavage of caspases, and TUNEL staining. NAC and Ac-DEVD-CHO partially reversed CuE-induced cleavage of caspase-3, caspase-7, and PARP. Furthermore, CuE caused accumulation of autophagic vacuoles and concentration- and time-dependent expression of LC3II protein. Autophagy inhibitors chloroquine and bafilomycin A1 enhanced CuE-induced LC3II expression and cell death. CuE-triggered protein expression of p-AKT, p-mTOR, Beclin-1, and p-ULK1 was partially reversed by NAC pretreatment. In addition, CuE treatment damaged F-actin without affecting β-tubulin as confirmed by immunofluorescence. In conclusion, CuE induced ROS-dependent apoptosis through Bcl-2 family and caspases in 95D lung cancer cells. Furthermore, CuE induced protective autophagy mediated by ROS through AKT/mTOR pathway. This study provides novel roles of ROS in the anticancer effect of CuE.
KW - Apoptosis
KW - Autophagy
KW - Cancer
KW - Cucurbitacin E
KW - Reactive oxygen species
UR - http://www.scopus.com/inward/record.url?scp=84964608743&partnerID=8YFLogxK
U2 - 10.1016/j.cbi.2016.04.028
DO - 10.1016/j.cbi.2016.04.028
M3 - Journal article
C2 - 27106530
AN - SCOPUS:84964608743
SN - 0009-2797
VL - 253
SP - 1
EP - 9
JO - Chemico-Biological Interactions
JF - Chemico-Biological Interactions
ER -