Nitric oxide and calcium ions in apoptotic esophageal carcinoma cells induced by arsenite.
OBJECTIVE
To Quantitatively analyze the nitric oxide (NO) and Ca2+ in apoptosis of esophageal carcinoma cells induced by arsenic trioxide (As2O3).


METHODS
The cell line SHEEC1, a malignant esophageal epithelial cell induced by HPV in synergy with TPA in our laboratory, was cultured in a serum-free medium and treated with As2O3. Before and after administration of As2O3, NO production in cultured medium was detected quantitatively using the Griess Colorimetric method. Intracellular Ca2+ was labeled by using the fluorescent dye Fluo3-AM and detected under confocal laser scanning microscope (CLSM), which was able to acquire data in real-time enabling Ca2+ dynamics of individual cells in vitro. The apoptotic cells were examined under electron microscopy.


RESULTS
Intracellular concentration of Ca2+ increased from 1.00 units to 1.09-1.38 units of fluorescent intensity at As2O3 treatment and NO products subsequently released from As2O3-treated cells increased from 0.98-1.00 x10(-2)micromol x L(-1) up to 1.48-1.52 x10(-2)micromol x L(-1) and maintained in a high level continuously. Finally apoptosis of cells occurred,chromatin being agglutinated, cells shrunk,nuclei became round and mitochondria swelled.


CONCLUSIONS
Ca2+ and NO increased with cell damage and apoptosis in cells treated by As2O3. The Ca2+ is an initial messenger to the apoptotic pathway. To investigate Ca2+ and NO will be a new direction for studying the apoptotic signaling messenger of the esophageal carcinoma cells induced by As2O3.
Shen
Zhong-Ying
ZY
Department of Pathology, Medical College of Shantou University, 22 Xinling Road, Shantou 515031, Guandong Province, China. zhongyingshen@yahoo.com
Shen
Wen-Ying
WY
Chen
Ming-Hua
MH
Shen
Jian
J
Cai
Wei-Jie
WJ
Yi
Zeng
Z
D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects / Q000502:physiology*; D000077237:Arsenic Trioxide; D001152:Arsenicals / Q000494:pharmacology; D018053:Arsenites / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism*; D004938:Esophageal Neoplasms; D006801:Humans; D008854:Microscopy, Electron; D009569:Nitric Oxide / Q000378:metabolism*; D010087:Oxides / Q000494:pharmacology; D013723:Teratogens / Q000494:pharmacology*; D014407:Tumor Cells, Cultured / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure
D000970:Antineoplastic Agents; D001152:Arsenicals; D018053:Arsenites; D010087:Oxides; D013723:Teratogens; D009569:Nitric Oxide; C015001:arsenite; D000077237:Arsenic Trioxide; D002118:Calcium