@#The aim of this study was to investigate the effect of norcantharidin (NCTD) on the proliferation and
apoptosis of
triple-negative breast cancer cell line MDA-MB-231.
Western blot was used to detect the effect of NCTD on the expression levels of
apoptosis-related
proteins Bax/Bcl-2, cleaved-PARP/PARP/PARP, cleved-
caspase-9, cleaved-
caspase-3 and MCL-1 in
MDA-MB-231 cells.Also, the expression levels of
autophagy-related proteins LC3-II/LC3-I, Parkin and PINK1 in
MDA-MB-231 cells were measured by
Western blot.
Flow cytometry was used to
measure the effect of NCTD on the changes of
mitochondrial membrane potential and mitochondrial
reactive oxygen species (ROS).The effect of NCTD on
autophagy flow in
cells expressing mCherry-EGFP-LC3 was detected by a confocal microscope.Moreover, the effects of NCTD combined with
chloroquine (CQ) or 3-methyladenine (3-MA) on the
apoptosis of
MDA-MB-231 cells were detected by
flow cytometry.The results showed that NCTD significantly increased the expression levels of Bax/Bcl-2, cleaved-PARP/PARP, cleaved-
caspase-9, cleasved-
caspase-3 and LC3-II/LC3-I
proteins, and promoted the mitochondrial translocation of Parkin, and blocked the autophagic flow in
MDA-MB-231 cells. Moreover, NCTD combined with CQ accelerated
apoptosis, while NCTD combined with 3-MA decreased
apoptosis.These results suggest that NCTD can induce
autophagy accumulation and
lead to
apoptosis of
MDA-MB-231 cells.