Miltirone induces cell death in hepatocellular carcinoma cell through GSDME-dependent pyroptosis.
10.1016/j.apsb.2020.06.015
- Author:
Xiaowei ZHANG
1
;
Ping ZHANG
1
;
Lin AN
1
;
Ningyuan SUN
1
;
Liying PENG
1
;
Weiwei TANG
1
;
Dingyuan MA
2
;
Jun CHEN
1
Author Information
1. State Key Laboratory of Natural Medicines, China Pharmaceutical University, Nanjing 210009, China.
2. Department of Prenatal Diagnosis, Nanjing Maternity and Child Health Care Hospital, Women's Hospital of Nanjing Medical University, Nanjing 210004, China.
- Publication Type:Journal Article
- Keywords:
7-AAD, 7-aminoactinomycin D;
AKT, AKT serine/threonine kinase, also known as protein kinase B;
ANOVA, analysis of variance;
BAX, BCL2-associated X;
CCK-8, cell counting kit-8;
CRISPR, clustered regularly interspaced short palindromic repeats;
Cas9, caspase 9;
Cell death;
DCFH-DA, dye 2,7-dichlorofluoresce diacetate;
DMEM, Dulbecco's modified Eagle's medium;
DMSO, dimethyl sulfoxide;
ECL, enhanced chemiluminescence;
ERK1/2, extracellular regulated protein kinases 1/2;
FBS, fetal bovine serum;
FITC, fluorescein isothiocyanate;
GAPDH, glyceraldehyde-3-phosphate dehydrogenase;
GSDMD, gasdermin D;
GSDME;
GSDME, gasdermin E;
H&E, hematoxylin and eosin;
HCC, hepatocellular carcinoma;
HRP, horseradish peroxidase;
HepG2;
Hepa1-6;
Hepatocellular carcinoma;
IC50, the half maximal inhibitory concentration;
IgG (H + L), immunoglobulin G (heavy chain + light chain);
KO, knockout;
LDH, lactic dehydrogenase;
MEK, mitogen-activated and extracellular signal-regulated kinase;
MEM, minimum essential medium;
MMP, mitochondrial membrane potential;
MS, mass spectrum;
Miltirone;
N-GSDME, N-terminal GSDME;
NAC, N-acetyl cysteine;
NC, negative control;
NMR, nuclear magnetic resonance;
NS, no significance;
PARP, poly ADP-ribose polymerase;
PBS, phosphate-based buffer;
PI, propidium iodide;
PI3K, phosphatidylinositol 3-kinase;
Pyroptosis;
RIPA, radioimmunoprecipitation assay;
ROS, reactive oxygen species;
SD, standard deviation;
SDS-PAGE, sodium dodecyl sulphate-polyacrylamide gel electrophoresis;
TBST, Tris-buffered saline with Tween solution;
TCGA, the Cancer Genome Atlas;
VEGF, vascular endothelial growth factor;
gRNA, guide RNA;
i.p., intraperitoneal;
i.v., intravenous;
mTOR, mammalian target of rapamycin;
p-AKT, phosphorylated-AKT;
p-ERK1/2, phosphorylated-ERK1/2;
p-MEK, phosphorylated-MEK
- From:
Acta Pharmaceutica Sinica B
2020;10(8):1397-1413
- CountryChina
- Language:English
-
Abstract:
Pyroptosis is a form of programmed cell death, and recently described as a new molecular mechanism of chemotherapy drugs in the treatment of tumors. Miltirone, a derivative of phenanthrene-quinone isolated from the root of Bunge, has been shown to possess anti-cancer activities. Here, we found that miltirone inhibited the cell viability of either HepG2 or Hepa1-6 cells, and induced the proteolytic cleavage of gasdermin E (GSDME) in each hepatocellular carcinoma (HCC) cell line, with concomitant cleavage of caspase 3. Knocking out switched miltirone-induced cell death from pyroptosis to apoptosis. Additionally, the induction effects of miltirone on GSDME-dependent pyroptosis were attenuated by siRNA-mediated caspase three silencing and the specific caspase three inhibitor Z-DEVD-FMK, respectively. Miltirone effectively elicited intracellular accumulation of reactive oxygen species (ROS), and suppressed phosphorylation of mitogen-activated and extracellular signal-regulated kinase (MEK) and extracellular regulated protein kinases 1/2 (ERK1/2) for pyroptosis induction. Moreover, miltirone significantly inhibited tumor growth and induced pyroptosis in the Hepa1-6 mouse HCC syngeneic model. These results provide a new insight that miltirone is a potential therapeutic agent for the treatment of HCC GSDME-dependent pyroptosis.
