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目的:深入研究力达霉素对人肝癌BEL-7402细胞基因组DNA的影响.方法:琼脂糖凝胶电泳检测DNA泳带;Southern杂交检测对基因的损伤;中性凝胶电泳检测DNA断裂修复;超螺旋GEM质粒检测断裂DNA位点.结果:力达霉素1μmol/L处理BEL-7402细胞15、30、60min,可检测到明显的DNA梯带.低浓度的力达霉素对活跃基因N-ras的外显子和内含子均有明显的损伤作用,但对沉默IL-2基因没有影响.力达霉素断裂DNA后,DNA损伤难以修复.力达霉素断裂GEM质粒DNA的位点是-OH HO-.结论:力达霉素明显地损伤人肝癌BEL-7402细胞的基因组DNA.该结果有助于解释其高活性地杀伤肿瘤细胞的分子机制.
OBJECTIVE: To study the effect of lidamycin on genomic DNA of human hepatocellular carcinoma BEL-7402 cells in vitro.METHODS: DNA bands were detected by agarose gel electrophoresis, DNA damage was detected by Southern blotting, DNA fragmentation was detected by neutral gel electrophoresis, DNA was detected by supercoiled GEM plasmid.Results: Lidamycin 1 μmol / L treated BEL-7402 cells for 15, 30 and 60 min, and significant DNA ladder could be detected.Low concentration of lidamycin on the active gene N -ras had significant damage in both exon and intron but had no effect on silencing of IL-2 gene.It is difficult to repair DNA damage by ligusticin, Point is -OH HO- .Conclusion: Lidamycin can significantly damage the genomic DNA of human hepatocellular carcinoma BEL-7402 cells.This result is helpful to explain its molecular mechanism of highly active killing of tumor cells.