摘要
目的研究顺铂对人葡萄膜黑色素瘤细胞端粒酶活性的抑制作用，为临床治疗人葡萄膜黑色素瘤提供理论依据。方法采用特定时间下不同浓度以及特定浓度下不同作用时间的端粒酶抑制剂顺铂作用于体外培养的人黑色素瘤细胞。采用多聚酶链反应——酶联免疫吸附测定（PCR—ELISA）及聚丙烯酰胺凝胶电泳法（PCR—PAGE）测定细胞中端粒酶活性的变化。结果作用72h，端粒酶活性在顺铂浓度达到0．10mg／L后开始下降，当浓度达到1．00mg／L后其活性下降至（0．173±0．007）。当顺铂浓度固定10．00mg／L，顺铂作用时间达到24h后开始出现抑制作用，48h时达到抑制高峰，端粒酶活性下降至（0．276±0．024）。随着顺铂浓度的增加及作用时间的延长，端粒酶活性逐渐下降（P〈0．05）。PCR－PAGE显示顺铂浓度增加及作用时间延长，端粒酶活性的显色条带越来越少。结论顺铂可有效降低体外培养的人眼葡萄膜黑色素瘤细胞端粒酶活性，并呈浓度和时间依赖性。
Objective To investigate the inhibitory effect of CDDP （Cisplatin, cisdichlorodiamine platinum） on telomerase activity in uveal melanoma cells of human eyes and to provide evidence for clinical treatment of uveal melanoma. Methods CDDP treatment with varying concentrations and definite time, and with various CDDP interference time and definite concentration, were applied to the uveal melanoma cells of human eyes cultured in vitro. The changes of telomerase activity in cells were detected by polymcrase chain reaction-enzyme linked immunosorbent assay （PCR-ELISA） and polymerasc chain reaction- polyscrylamide gel electrophoresis （PCR- PAGE）. Results With 72 hours treatment, telomerase activity began to decrease with 0.10 mg/L CDDP and reached the lowest level （0.173±0.007） with 1.00 mg/L CDDP. With a definite concentration （ 10.00 mg/L CDDP）, telomerase activity was inhibited after 24 hours, and reached the lowest level （0.276±0.024） after 48 hours. Namely, along with higher level of CDDP and longer time of treatment, telomerase activity was reduced （P 〈 0.05）, and fewer color bands of telomerase activity was visualized on PCR-PAGE. Conclusion CDDP can effectively inhibit the tolemerase activity of uveal melanoma cells of human eyes cultured in vitro in a concentration- and time-dependent manner.
出处
《中华生物医学工程杂志》
CAS
2010年第3期227-230,共4页
Chinese Journal of Biomedical Engineering
基金
基金项目：广东省科技计划项目（20088030301253）
关键词
黑色素瘤
脉络膜肿瘤
端粒酶
顺铂
多聚酶链反应
Telomcrase
Choroid neoplasms
Melanoma
Cisplatin
Polymerase chain reaction
