论文部分内容阅读
目的研究中药大黄素(emodin)对人髓系白血病细胞株HL-60细胞的影响及探讨cmyc基因在其中的作用。方法应用MTT法绘制细胞生长曲线;细胞集落培养观察大黄素对HL-60细胞增殖的影响;DNA倍体分析、线粒体细胞凋亡检测法、末端缺失原位标记(TUNEL)法及DNA凝胶电泳分析细胞凋亡;RTPCR及Westernblot检测大黄素作用前后cmyc基因mRNA及蛋白表达水平的变化。结果大黄素能明显抑制HL-60细胞增殖,半数抑制浓度(IC50)约为20μmol/L;DNA片段化、亚G1峰(凋亡峰)的检出及线粒体细胞凋亡检测等证实大黄素能有效诱导HL-60细胞凋亡,细胞凋亡率与药物作用浓度呈正相关。大黄素与HL-60细胞作用后cmyc基因mRNA及蛋白的表达水平与作用时间呈负相关。结论大黄素能有效抑制HL-60细胞增殖,诱导其凋亡;cmyc基因可能参与了大黄素抑制HL60细胞增殖和诱导凋亡的过程。
Objective To study the effect of emodin on human myeloid leukemia cell line HL-60 and explore the role of cmyc gene in it. Methods The cell growth curve was drawn by MTT method; the effect of emodin on the proliferation of HL-60 cells was observed by cell colony culture; DNA ploidy analysis, mitochondrial apoptosis assay, terminal deletion TUNEL assay and DNA gel electrophoresis The cell apoptosis was analyzed. RT-PCR and Western blot were used to detect the changes of mRNA and protein expression of cmyc gene before and after emodin treatment. Results Emodin could significantly inhibit the proliferation of HL-60 cells with an IC50 of about 20 μmol/L. DNA fragmentation, sub-G1 peak (apoptosis peak) detection and mitochondrial apoptosis detection confirmed emodin. The apoptosis of HL-60 cells was effectively induced, and the apoptosis rate was positively correlated with the concentration of drug. The expression level of mRNA and protein of cmyc gene was negatively correlated with the action time after emodin treatment with HL-60 cells. Conclusion Emodin can effectively inhibit the proliferation of HL-60 cells and induce apoptosis. The CMYC gene may participate in the process of emodin inhibiting the proliferation of HL60 cells and inducing apoptosis.