单细胞凝胶电泳技术在DNA损伤和细胞凋亡检测中的应用

APPLICATION OF THE SINGLE CELL GEL ELECTROPHORESIS IN THE DETECTION OF DNA DAMAGE AND APOPTOSIS

用不同质量浓度的丝裂霉素C(MMC)、卡铂和5-氟尿嘧啶(5-Fu)处理Hela细胞24 h,利用单细胞凝胶电泳(SCGE)技术分析这些药物对细胞DNA的损伤情况,结果发现:各药物处理组细胞都出现"彗星",且随着各药物浓度的增大,彗星率升高,彗尾的荧光强度和长度也随之增大;各剂量组间存在良好的剂量-效应关系,差异显著(P<0.05).3种药物对Hela细胞DNA均有一定程度的损伤.单细胞凝胶电泳可用于快速检测哺乳动物细胞DNA损伤和细胞凋亡.

The single cell gel electrophoresis（SCGE） was performed to detect the comet proportion,the fluorescence intensity and length of comet tail after Hela cells were treated for 24 hours by different concentrations of three kinds of anticancer drugs,mitomycin C（MMC）,carboplatin and 5-fluorouracil（5-Fu）,and the feasibility of SCGE in the detection of apoptosis was investigated.The results showed that all of the treated groups showed comets after SCGE.The comet proportion increased as the concentration of drug increased,and so did the fluorescence intensity and length of comet tail.There was good dose-response relationships in each drug and the differences between different groups were significant（P0.05）.These results indicated that all of three kinds of anticancer drugs induce the nuclear DNA damage in Hela cells,and SCEG can be used as a simple,rapid and sensitive method to detect DNA damage and cell apoptosis.