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目的:探讨多柔比星(ADM)诱导肺腺癌细胞系A549细胞凋亡机制。方法:应用噻唑蓝(MTT)比色法检测ADM对A549细胞增殖抑制作用,透射电镜观察细胞形态学改变;细胞电泳仪测定A549细胞表面电荷的变化;流式细胞仪(FCM)测定A549细胞Fas、Bcl-2蛋白水平变化;比色法检测Caspase-3活性的变化。结果:ADM可抑制细胞增殖,作用24、48及72h的IC50值分别为5.2、4.8和4.2mg/L,电镜下见到明显的凋亡细胞;细胞表面电位在4h开始下降,4～24h期间下降最明显,>24h下降趋势降低;Fas蛋白表达明显上调,Bcl-2蛋白表达下降,Caspase-3活性明显增强。结论:ADM能抑制并诱导A549细胞凋亡,细胞表面电荷下降可能是ADM诱导细胞凋亡的早期事件,其作用可能与上调Fas蛋白、下调Bcl-2蛋白表达有关。
Aims: To investigate the mechanism of apoptosis induced by doxorubicin in human lung adenocarcionoma cell line A549. Methods: MTT assay was used to detect the inhibitory effect of ADM on the proliferation of A549 cells. The morphological changes of cells were observed by transmission electron microscopy. The changes of surface charge of A549 cells were determined by cell electrophoresis. The expression of Fas in A549 cells was detected by flow cytometry (FCM) , Bcl-2 protein level changes; colorimetric assay Caspase-3 activity changes. Results: ADM could inhibit cell proliferation. The IC50 values at 24, 48 and 72 h were 5.2, 4.8 and 4.2 mg / L, respectively. The apoptotic cells were observed under electron microscope. The cell surface potential began to decrease at 4 h. The most obvious decline,> 24h decline trend decreased; Fas protein expression was significantly increased, decreased Bcl-2 protein expression, Caspase-3 activity was significantly enhanced. CONCLUSION: ADM can inhibit and induce the apoptosis of A549 cells. The decrease of cell surface charge may be an early event of apoptosis induced by ADM. Its effect may be related to the up-regulation of Fas protein and down-regulation of Bcl-2 protein expression.