[The role of reactive oxygen species in cisplatin-induced apoptosis of esophageal cancer cell line EC-109].
OBJECTIVE
Reactive oxygen species (ROS), in vivo oxygen metabolites and important signaling molecules, play a vital role in cell apoptosis. This study was to investigate the role of ROS in cisplatin (DDP)-induced apoptosis of esophageal cancer cell line EC-109, and explore the mechanism.


METHODS
EC-109 cells were treated with different concentrations (0, 1, 5, 10, and 15 microg/ml) of DDP. MTT assay was used to evaluate the influence of DDP on cell proliferation. Flow cytometry was used to test ROS levels, intracellular mitochondrial transmembrane potential (Delta psi m), and hypodiploid apoptosis peak in EC-109 cells. Cell apoptosis after pretreatment with hydrogen peroxide-scavenging enzyme catalase (CAT) was also detected.


RESULTS
DDP obviously suppressed proliferation of EC-109 cells. When treated with 0, 1, 5, 10, 15 microg/ml of DDP for 2 h, ROS levels were (3.3+/-1.0)%, (21.6+/-2.0)%, (32.6+/-3.2)%, (44.7+/-2.2)%, and (53.1+/-3.6)%, respectively; when treated for 12 h, Delta psi m were (97.2+/-1.9)%, (90.6+/-1.9)%, (85.5+/-1.4)%, (67.8+/-2.0)%, and (62.4+/-3.0)%, respectively; when treated for 24 h, cell apoptotic rates were (3.4+/-1.2)%, (16.2+/-2.3)%, (28.1+/-1.5)%, (33.2+/-3.9)%, and (45.5+/-3.8)%, respectively. Pretreatment with CAT significantly rescued cells from apoptosis (P<0.05).


CONCLUSIONS
DDP generates ROS in esophageal cancer EC-109 cells, which causes mitochondrial membrane permeabilization and Delta psi m decrease, therefore, leads to apoptosis of EC-109 cells.
Cai
Xian-Bin
XB
Department of Gastroenterology, The First Affiliated Hospital, Medical College, Shantou University, Shantou, Guangdong 515041, P. R. China. g_xbcai@stu.edu.cn
Jing
Xu-Bin
XB
Hu
Hui
H
Chen
Su-Zuan
SZ
Li
Ying-Hua
YH
D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects*; D002374:Catalase / Q000494:pharmacology; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002945:Cisplatin / Q000008:administration & dosage / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004938:Esophageal Neoplasms / Q000378:metabolism / Q000473:pathology*; D005434:Flow Cytometry; D006801:Humans; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D017382:Reactive Oxygen Species / Q000378:metabolism*
D000970:Antineoplastic Agents; D017382:Reactive Oxygen Species; D002374:Catalase; D002945:Cisplatin