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目的探讨曲古抑菌素A(trichostatin A,TSA)对顺铂诱导的细胞毒性的影响。方法给予0、250、500 nmol/L TSA预处理A549细胞24 h后,分别联合0、5、10、15μmol/L顺铂处理A549细胞24 h后,通过0、0.5、1、2μmol/L APE1 InhibitorⅢ调控APE1 BER功能,应用CCK-8方法检测细胞增殖率,AP内切酶活性实验检测APE1 BER功能,通过流式细胞仪检测细胞凋亡水平,immunoblot检测γ-H2AX表达水平。结果 TSA联合顺铂处理A549细胞明显增强顺铂的细胞增殖抑制,并促进顺铂诱导的细胞凋亡;随着TSA浓度的升高APE1内切酶活性逐渐减弱,TSA联合顺铂促进顺铂诱导的γ-H2AX表达;利用inhibitorⅢ抑制APE1内切酶活性,促进顺铂诱导细胞增殖抑制和凋亡水平。结论TSA可能通过抑制APE1BER功能增强顺铂诱导的细胞毒性反应。
Objective To investigate the effect of trichostatin A (TSA) on cisplatin-induced cytotoxicity. Methods A549 cells were pretreated with 0,250 and 500 nmol / L TSA for 24 h. A549 cells were treated with 0, 5, 10 and 15 μmol / L cisplatin respectively for 24 h and then treated with 0, 0.5, 1 and 2 μmol / L APE1 Inhibitor III regulated the function of APE1 BER. The cell proliferation rate was detected by CCK-8 assay. The function of APE1 BER was detected by AP endonuclease activity assay. The level of apoptosis was detected by flow cytometry. The expression of γ-H2AX was detected by immunoblot. Results TSA combined with cisplatin treatment of A549 cells significantly enhanced cisplatin inhibition of cell proliferation and promote cisplatin-induced apoptosis; as the concentration of TSA APE1 endonuclease activity gradually weakened, TSA combined with cisplatin promote cisplatin-induced Γ-H2AX. Inhibitor III could inhibit the activity of APE1 endonuclease and promote cisplatin-induced cell proliferation and apoptosis. Conclusion TSA may enhance cisplatin-induced cytotoxicity by inhibiting APE1BER function.