Carbenoxolone enhances inhibitory effect of RSL3 against cisplatin-resistant testicular cancer cells by promoting ferroptosis.
10.12122/j.issn.1673-4254.2022.03.13
- Author:
Jia Ru DU
1
;
Bin LI
1
;
Chen Lu ZHU
1
;
Jia Le HAN
1
;
Xu Hui TONG
1
Author Information
1. School of Pharmacy, Bengbu Medical College, Anhui Provincial Engineering Research Center for Biochemical Pharmaceuticals, Bengbu 233030, China.
- Publication Type:Journal Article
- Keywords:
RSL3;
carbenoxolone;
ferroptosis;
invasion;
migration;
proliferation;
testicular cancer
- MeSH:
Carbenoxolone/pharmacology*;
Cell Line, Tumor;
Cell Proliferation;
Cisplatin/pharmacology*;
Ferroptosis;
Fluorescent Dyes/pharmacology*;
Humans;
Lipids;
Male;
Neoplasms, Germ Cell and Embryonal;
Reactive Oxygen Species;
Testicular Neoplasms
- From:
Journal of Southern Medical University
2022;42(3):405-410
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the inhibitory effect of RSL3 on the proliferation, invasion and migration of cisplatinresistant testicular cancer cells (I-10/DDP) and the effect of carbenoxolone on the activity of RSL3 against testicular cancer.
METHODS:MTT assay was used to evaluate the survival rate of I-10/DDP cells following treatment with RSL3 (1, 2, 4, 8, 16 or 32 μmol/L) alone or in combination with carbenoxolone (100 μmol/L) or after treatment with Fer-1 (2 μmol/L), RSL3 (4 μmol/L), RSL3+Fer-1, RSL3+carbenoxolone (100 μmol/L), or RSL3+Fer-1+carbenoxolone. Colony formation assay was used to assess the proliferation ability of the treated cells; wounding-healing assay and Transwell assay were used to assess the invasion and migration ability of the cells. The expression of GPX4 was detected using Western blotting, the levels of lipid ROS were detected using C11 BODIPY 581/591 fluorescent probe, and the levels of Fe2+ were determined with FerroOrange fluorescent probe.
RESULTS:RSL3 dose-dependently decreased the survival rate of I-10/DDP cells, and the combined treatment with 2, 4, or 8 μmol/L RSL3 with carbenoxolone, as compared with RSL3 treatment alone, resulted in significant reduction of the cell survival rate. The combination with carbenoxolone significantly enhanced the inhibitory effect of RSL3 on colony formation, wound healing rate (P=0.005), invasion and migration of the cells (P < 0.001). Fer-1 obviously attenuated the inhibitory effects of RSL3 alone and its combination with carbenoxolone on I-10/DDP cells (P < 0.01). RSL3 treatment significantly decreased GPX4 expression (P=0.001) and increased lipid ROS level (P=0.001) and Fe2+ level in the cells, and these effects were further enhanced by the combined treatment with carbenoxolone (P < 0.01).
CONCLUSION:Carbenoxolone enhances the inhibitory effect of RSL3 on the proliferation, invasion and migration of cisplatin-resistant testicular cancer cells by promoting RSL3-induced ferroptosis.
