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目的研究柔红霉素诱导K562细胞增殖和凋亡情况,以及调节细胞周期和FAKmRNA基因,进一步探讨通过调控FAK表达,研究抗白血病的作用机制。方法应用CCK8细胞增殖法,结合流式细胞仪检测法检测不同浓度柔红霉素在不同时间对K562细胞增殖和凋亡的影响,应用RT-PCR和Westernblot技术检测不同浓度柔红霉素作用对K562细胞FAKmRNA以及蛋白表达水平的变化。结果 K562细胞的增殖抑制率随着柔红霉素浓度增加及作用时间延长逐渐升高,同一浓度不同时间组之间,或者同一时间不同浓度组之间比较,差异均有统计学意义(P<0.05);柔红霉素能引起K562细胞凋亡,且随着药物浓度增加,凋亡率也逐渐增加,差异均有统计学意义(P<0.05);柔红霉素能引起K562细胞周期阻滞,多停留在S期;柔红霉素能引起K562细胞FAKmRNA表达和FAK蛋白表达水平的降低。结论一定浓度的柔红霉素在体外可诱导细胞凋亡增加并抑制分裂期细胞的增殖,对细胞FAK基因和蛋白水平都有显著下调,发生凋亡的机制可能是通过抑制FAK基因表达。
Objective To study the proliferation and apoptosis of K562 cells induced by daunorubicin, and to study the regulation of cell cycle and FAK mRNA gene expression. To further investigate the mechanism of anti-leukemia by regulating FAK expression. Methods CCK8 cell proliferation assay and flow cytometry assay were used to detect the effects of different concentrations of daunorubicin on proliferation and apoptosis of K562 cells at different times. RT-PCR and Western blotting were used to detect the effect of daunorubicin K562 cells FAKmRNA and protein expression levels. Results The proliferation inhibition rate of K562 cells increased gradually with the increase of daunorubicin concentration and the prolongation of action time. The difference was statistically significant between different time groups of the same concentration or different concentration groups at the same time (P < 0.05). Daunorubicin induced apoptosis in K562 cells, and the apoptotic rate also increased gradually with the increase of drug concentration (P <0.05). Daunorubicin could induce cell cycle arrest in K562 cells Hysteresis, and more stay in the S phase; daunorubicin K562 cells can cause FAKmRNA expression and FAK protein expression decreased. CONCLUSION: A certain concentration of daunorubicin can induce apoptosis in vitro and inhibit the proliferation of mitotic cells, and significantly decrease the expression of FAK gene and protein. The mechanism of apoptosis may be through inhibition of FAK gene expression.