In Vitro Study on Lethal Effect of Human Choroidal Melanoma OCM-1 Cell Line by Repeating -70℃ Freeze Thawing  

作　　者：Bin Liu Yongping Li Bo Zhang Wenxin Zhang 

机构地区：[1]National Key Laboratory of Ophthalmology, Zhongshan Ophthalmic Center, Sun Yat-sen University Guangzhou 510060, China

出　　处：《眼科学报》2007年第1期31-36,共6页Eye Science

基　　金：Foundation item: National Natural Science Foundation of China(No. 39870801, 30672276);society development item of Guangdong province(No. 2003A3020302)

摘　　要：Objective: To investigate the effects of repeating-70℃freeze thawing on human choroidal melanoma cell line OCM-1. Methods: OCM-1 cells were frozen by repeating -70℃ freeze thawing with various durations and frequencies. Then the inhibit rate of cells was examined by MTT essay. The cell viability was measured by monoclonal formation assay. We also used the HE staining, immunohistochemistry staining and the laser-scanning confocal microscopy (LSCM) to investigate the morphological changes of the cells. Results: The growth of OCM-1 cells was inhibited by repeating -70℃ freeze thawing in time-dependent and frequency-dependent manners (P < 0.01). Different morphous including necrosis and apoptosis of the cells could be observed after -70℃ freeze thawing by the LSCM. Conclusion: Repeating -70℃ freeze thawing can not only kill cells directly and induce considerable cells to apoptosis, but also inhibit the growth of the survivals. The kill and wound ratio of the cells disposal with different times and frequencies present variance. And the distinction when treated with different frequencies during the same time is much more significant than different times with the same frequency, which guide clinical workers to choose repeating cryotherapy with short term method instead of single cryotherapy with long term in choroidal melanoma treatment.Objective： To investigate the effects of repeating -70℃ freeze thawing on human ehoroidal melanoma cell line OCM-1. Methods： OCM-1 cells were frozen by repeating -70℃ freeze thawing with various durations and frequencies. Then the inhibit rate of cells was examined by MTT essay. The cell viability was measured by monoclonal formation assay. We also used the HE staining, immunohistochemistry staining and the laser-scanning confocal microscopy （LSCM） to investigate the morphological changes of the cells. Results： The growth of OCM-1 cells was inhibited by repeating -70℃ freeze thawing in time-dependent and frequency-dependent manners （P 〈 0.01）. Different morphous including necrosis and apoptosis of the cells could be observed after -70℃ freeze thawing by the LSCM. Conclusion： Repeating -70℃ freeze thawing can not only kill cells directly and induce considerable cells to apoptosis, but also inhibit the growth of the survivals. The kill and wound ratio of the cells disposal with different times and frequencies present variance. And the distinction when treated with different frequencies during the same time is much more significant than different times with the same frequency, which guide clinical workers to choose repeating cryotherapy with short term method instead of single cryotherapy with long term in choroidal melanoma treatment.

关 键 词：冻融 脉络膜黑素瘤 OCM-1细胞 杀伤效果 体外研究 

分 类 号：R77[医药卫生—眼科]