Comparative toxicity and apoptosis induced by diorganotins in rat pheochromocytoma (PC12) cells.
As ubiquitous environmental toxicants, organotin (IV) compounds (OTC) accumulate in the food chain and potential effects on human health are disquieting. The present study compared the cytotoxicity of three diorganotins, namely, dimethyltin (DMT), dibutyltin (DBT) and diphenyltin (DPT), in rat pheochromocytoma (PC12) cells, and the molecular mechanisms responsible for their cytotoxic effects were also explored. Twenty-four hours exposure of PC12 cells to DBT and DPT resulted in a concentration-dependent decrease in cell viability with median lethal concentration (LC₅₀) of 2.97 μM and 7.24 μM, respectively. However, DMT at concentrations up to 128 μM had no obvious effect on cell viability. The mechanistic study revealed that the extent of apoptosis was greater for DBT than that for DPT, followed by DMT, as evidenced by acridine orange/ethidium bromide (AO/EB) fluorescent staining method and annexin V-FITC/PI staining flow cytometry analysis, as well as generation of intracellular reactive oxygen species (ROS), mitochondrial membrane potential (MMP) disruption, release of cytochrome c (Cyt c), and consequent activation of caspase-9, and -3. These investigations suggested that the cytotoxic potency of three diorganotins in PC12 cells was in the order of DBT>DPT≫DMT, and these compounds could induce PC12 cells apoptosis through ROS mediated mitochondrial pathway.
School of Pharmaceutical Science, Shanxi Medical University, No. 56, Xinjian Nan Road, Taiyuan 030001, Shanxi, People's Republic of China.
Enli
E
Liu
Xue
X
Du
Rui
R
Ge
Taigang
T
Liang
Qiao
Q
Niu
Qingshan
Q
Li
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D053148:Caspase 3 / Q000235:genetics / Q000378:metabolism; D053453:Caspase 9 / Q000235:genetics / Q000378:metabolism; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000187:drug effects; D045304:Cytochromes c / Q000378:metabolism; D007928:Lethal Dose 50; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D009947:Organotin Compounds / Q000633:toxicity*; D016716:PC12 Cells; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism
D009947:Organotin Compounds; D017382:Reactive Oxygen Species; C056189:di-n-butyltin; C063612:diphenyltin; C039760:dimethyltin; D045304:Cytochromes c; C505410:Casp3 protein, rat; C506435:Casp9 protein, rat; D053148:Caspase 3; D053453:Caspase 9
2′,7′-dichlorofluorescein-diacetate; 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid; 5,5′,6,6′-tetrachloro-1,1′,3,3′-tetraethyl-imidacarbocyanine iodide; AIF; AO; Apaf-1; Apoptosis; Cyt c; DBT; DCF; DCFH; DCFH-DA; DMEM; DMT; DPT; DTT; Dibutyltin (DBT); Dimethyltin (DMT); Diphenyltin (DPT); Dulbecco’s modified Eagle’s medium; EB; ECL; FBS; FITC; HEPES; HRP; JC-1; LC(50); MMP; MTT; Neurotoxicity; OTC; PC12 cells; PI; PMSF; PVC; ROS; TBS-T; TMT; acridine orange; apoptosis-inducing factor; apoptotic protease activation factor 1; cytochrome c; dibutyltin; dichlorofluorescein; dimethyltin; diphenyltin; dithiothreitol; ethidium bromide; fetal bovine serum; fluorescein isothiocyanate; horseradish peroxidase; median lethal concentration; mitochondrial membrane potential; organotin (IV) compounds; phenylmethanesulfonyl fluoride; polyvinyl chloride; propidium iodide; reactive oxygen species; the enhanced chemiluminescence; trimethyltin; tris-buffered saline plus Tween-20