中文摘要：

目的：研究人眼脉络膜黑色素瘤OCM．1细胞经深低温反复冻融后细胞内钙离子的变化及其与凋亡的关系。 方法：OCM-1细胞经-70℃冻融后，流式细胞仪检测细胞的凋亡比率，激光共聚焦显微镜观察细胞的形态学改变并检测细胞内钙离子浓度。 结果：OCM—1细胞冻融1，2，3次后，流式细胞仪显示细胞发生较高比例的凋亡，且随着冻融次数的增加而升高，凋亡比率（％）分别为12．4±1．4，30．3±1．0，38．0±1．9。共聚焦显微镜观察到细胞内钙离子荧光强度随着细胞凋亡比率的增高而升高，细胞内钙离子荧光强度分别为938．5±42．8，974．5±36．9，1362．8±73．3。 结论：深低温冻融能诱使OCM-1细胞内钙离子浓度发生改变，从而导致肿瘤细胞凋亡。

英文摘要：

AIM： To explore the effect of freeze thawing on the changes of intracellular [Ca^2±]i in human choroid melanoma OCM-1 cells, and investigate the relationship between the changes of intracellular [Ca^2±]i and apoptosis. METHODS： OCM-1 cells were frozen by repeating -70℃ freeze thawing for various frequency, then the cell apoptosis rates were examined by flow cytometry （FCM）, the morphological changes of the coils and the level of Ca^2± of the OCM-1 cells were examined by laser-scanning confocal microscopy （LSCM）. RESULTS： Freeze thawing could induco apoptosis of OCM-loells. The dassic morphologic characteristics of apoptosis could be seen. The different coil apoptosis rates were showed by FCM, and the level of Ca^2± of the OCM-1 coils were observed by LSCM. Both coil apoptosis rates and the levels of Ca^2± of the OCM-1 coils were advanced by increasing freeze thawing frequency. Freeze thawing 1, 2, 3 times, respectively, coil apoptosis rates （%） were 12.4± 1.4,30.3± 1.0,38.0±1.9 and the fluorescenoe intensity of intraoellular calcium ion were 938.5± 42.8, 974.5± 36.9, 1 362.8± 73.3. OONCLUSION： Freeze thawing can induoe the increasing of intraoellular [Ca^2±]i in vitro, then, led OCM-1 oells to apoptosis. The changes of [Ca^2±]i is the one of the important mechanisms of apoptosis.