双氢青蒿素联合顺铂活化Caspase-6、Lamin A/C表达诱导H1299细胞凋亡

Combination of dihydroartemisinin and cisplatin induced apoptosis in H1299 cells by cleaving Caspase-6 and Lamin A/C

目的研究双氢青蒿素(DHA)联合顺铂(Cis)对肺腺癌细胞株H1299的生长抑制作用效果及对半胱氨酸天冬氨酸蛋白酶6(Caspase-6)和核纤层蛋白A/C(Lamin A/C)表达的影响.方法体外培养人类肺腺癌H1299细胞并将细胞分为4组:DHA组、Cis组、DHA联合Cis组及对照组,通过四甲基偶氮唑盐/噻唑蓝实验来评价不同处理对H1299细胞增殖抑制的影响.采用流式细胞术检测以上各组对H1299细胞凋亡率的影响.通过赫斯特33258对细胞核完成染色来观察各组H1299细胞核的形态变化.采用蛋白质印迹法评价凋亡相关蛋白Caspase-6及Lamin A/C的表达情况.结果 DHA联合Cis组对H1299细胞的抑制率高于单药组.DHA联合Cis产生协同抑制H1299细胞的作用.DHA联合Cis对H1299细胞产生促凋亡作用.DHA联合Cis组Caspase-6及Lamin A/C的活性片段的表达均高于单独用药组.结论 DHA联合Cis组对肿瘤细胞的抑制明显高于单药组,并具有协同增强作用,其机制可能与提高了Caspase-6及Lamin A/C活性片段的表达有关.

Objective To explore the grow th inhibition effect of dihydroartem isinin (DHA) combined with Cisplatin (C is) and the expression of Caspase-6 and Lamin A /C to human lung adenocarcinoma H I 299 cells. Methods The cells were cultured in vitro and divided into four gro u p s: DHA group, Cis group, DHA plus Cis group and control group. Cell inhibition rate were evaluated by thiazolyl blue tetrazolium bromide assays. Flow cytom etry was used to detect the apoptosis induced by different treatm ent groups. Nuclear staining with Hoechst 33258 showed morphology of the nucleus treated with different cultivation m ethods. The apoptotic-associated proteins Caspase-6 and Lamin A /C were assessed by W estern blot. Results Inhibition rate of H I 299 cell line by D H A and Cis treated in combination was significantly higher than that in single drug groups. Combination of DHA and Cis had synergistic inhibitory effect on H1299 cell line. Combination of D H A and Cis had pro-apoptotic effect on H1299 cell line. The expressions of cleaved-Caspase-6 and cleaved-Lamin A /C by DHA and Cis treated in combination were higher than those in other groups. Conclusions T his study revealed the inhibition of the H1299 cells in the D HA plus Cis group was significantly higher than that in the single treatm ent group by increasing the expression of cleaved-Caspase-6 and cleaved-Lamin A /C .