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AIM:To quantitatively analyze the nitricokide(NO)andCa~(2+)in apoptosis of esophageal carcinoma cells induced byarsenic trioxide(As_2O_3).METHODS:The cell line SHEEC1,a malignant esophagealepithelial cell induced by HPV in synergy with TPA in ourlaboratory,was cultured in a serum-free medium and treatedwith As_2O_3.Before and after administration of As_2O_3,NOproduction in cultured medium was detected quantitativelyusing the Griess Colorimetric method.Intracellular Ca~(2+)waslabeled using the fluorescent dye Fluo3-AM and detectedunder confocal laser scanning microscope(CLSM),whichwas able to acquire data in real-time enabling Ca~(2+)dynamicsof individual cells in vitro.The apoptotic cells wareexamined under electron microscopy.RESULTS:Intracellular concentration of Ca~(2+)increased from1.00 units to 1.09-1.38 units of fluorescent intensity at As_2O_3treatment and NO products subsequently released fromAs_2O_3-treated cells increased from 0.98-1.00×10~(-2)mol·L~(-1)upto 1.48-1.52×10~(-2)mol·L~(-1)and maintained in a highlevel contineously.Finally apoptosis of cells occurred,chromatin being agglutinated,cells shrunk,nuclei becameround and mitochondria swelled.CONCLUSION:Ca~(2+)and NO increased with cell damage andapoptosis in cells treated by As_2O_3.The Ca~(2+)is an initialmessenger to the apoptotic pathway.To investigate Ca~(2+)and NO will be a new direction for studying the apoptoticsignaling messenger of the esophageal carcinoma cellsinduced by As_2O_3.
AIM:To quantitatively analyze the nitricokide(NO)andCa~(2+)in apoptosis of esophageal carcinoma cells induced byarsenic trioxide(As_2O_3).METHODS:The cell line SHEEC1,a malignant esophagealepithelial cell induced by HPV in synergy with TPA in our laboratory, Was cultured in a serum-free medium and treated with As_2O_3.Before and after administration of As_2O_3,NOproduction in cultured medium was detected quantitativelyusing the Griess Colorimetric method.Intracellular Ca 2+ was labeled with the fluorescent dye Fluo3-AM and detectedunder confocal laser Scanning microscope (CLSM), which was able to acquire data in real-time Ca~(2+)dynamics of individual cells in vitro. The apoptotic cells wareexamined under electron microscopy.RESULTS: Intracellular concentration of Ca~(2+) increased from 1. 00 units to 1.09-1.38 units of fluorescent intensity at As_2O_3treatment and NO productsresults releasedAs2O_3-treated cells increased from 0.98-1.00×10-2 mol·L-1 to upto 1.48-1.52×10~ -2)mol·L -1 and maintained in a highlevel contineously.Finally apoptosis of cells occurred, chromatin being agglutinated, cells shrunk,nuclei made rounded and mitochondria swelled.CONCLUSION:Ca 2+ and NO increased with cell Damage andapoptosis in cells treated by As_2O_3.The Ca~(2+)is an initialmessenger to the apoptotic pathway.To investigate Ca~(2+)and NO will be a new direction for studying the apoptoticsignaling messenger of the esophageal carcinoma cellsinduced by As_2O_3 .