顺铂对人脉络膜黑色素瘤细胞端粒酶活性影响的实验研究

Effects of dsplatin on telomerase activity of human choroidal melanoma cells

目的研究顺铂对人脉络膜黑色素瘤细胞端粒酶活性的影响及其对细胞的毒性作用，并探讨两者之间的关系，为临床化疗提供更多依据。方法实验研究。采用原代培养的人脉络膜黑色素瘤细胞，用特定时间下（72h）不同浓度（100．00、10．00、1．00、0．10、0．01mg／L）以及特定浓度下（10mg／L）不同时间（96、72、48、24、12h）的端粒酶抑制剂顺铂作用于黑色素瘤细胞，以生理盐水作用于细胞为空白对照组，用四甲基偶氮唑盐法检测药物对细胞的毒性作用，聚合酶链反应一酶联免疫吸附法测定细胞中端粒酶活性的变化。各组细胞端粒酶活性的变化采用单因素方差分析，细胞抑制率与端粒酶活性之间的关系采用Spearman等级相关分析。结果随着顺铂浓度的增加及作用时间的延长，细胞中端粒酶活性逐渐下降，在药物浓度〉0．10mg／L及作用时间达到24h后，其活性出现大幅度下降，随后出现细胞抑制率明显升高，与端粒酶活性下降呈负相关（r=-0．900，P=0．037），但前者均发生不同程度的滞后现象。结论顺铂作为一种端粒酶抑制剂，可有效降低体外培养的人脉络膜黑色素瘤细胞的端粒酶活性，呈浓度和时间依赖性，并造成细胞死亡。

Objective To study the effects of CDDP （cis-dichlorodiamine platinum） on the telomerase of human choroidal melanoma cells and to investigate the toxic effects of CDDP on these cells. To study the relationship between these two effects and to explore the possibility of using CDDP in the chemotherapy of choroidal melanoma. Methods It was an experimental research. CDDP was added to the culture medium of primary cultured human choroidal melanoma cells at different concentrations （0. 01,0. 1, 1, 10 and 100 rag/L, 72 h）and times （ 12, 24, 48, 72 and 96 h, 10 mg/L） and the results were compared with that of the controls. Toxic effects of CDDP were evaluated by MTT test and the level of telomerase was measured by PCR-ELISA assay. The correlation between these two effects was analyzed. Results The telomerase activity was inhibited by CDDP time dependently and dose dependently. Cell viability was decreased when the concentration of CDDP attained 0. 1 mg/L and acted for 24 hours. The cell toxicity of CDDP was correlated negatively （ r = - 0. 900, P = 0. 037 ） with the inhibition of telomerase. The cell death was lagged behind the decrease of telomerase. Conclusions CDDP is an effective telomerase inhibitor which can decrease the telomerase activity of cultured human choroidal melanoma cells significantly. This effect is dose and time dependent. CDDP can also cause the death of cultured melanoma cells.