目的 探讨伊曲康唑(itraconazole,Itra)抑制肺癌A549细胞增殖的作用及其机制。方法 Itra处理肺癌A549细胞24、48、96 h,用MTT法检测细胞存活率,并计算半抑制浓度(IC50)。将实验分为对照组、Itra组、AMPK抑制剂(compound C,Comp C)+Itra组,内质网应激抑制剂(4-PBA)+Itra组,用AnnexinV/PI法检测细胞凋亡,Western blot检测CHOP、GRP78、PERK、AMPK、磷酸化AMPK(p-AMPK)、Bax和Bcl-2的表达。结果 Itra显著降低A549细胞的存活率,在24、48、96h时的IC50分别为62.89、28.34L、11.64 μmol·L-1。与对照组相比,Itra组细胞存活率明显降低,细胞凋亡率明显增加(P＜0.05)。与Itra组相比,Comp C +Itra组和4-PBA+Itra组细胞存活率明显增加、凋亡率明显降低(P＜0.05)。与对照组相比,Itra组CHOP、GRP78、PERK、p-AMPK表达增加;与Itra组相比,Comp C +Itra组和4-PBA+ Itra组CHOP、GRP78、PERK、p-AMPK表达降低(P＜0.05)。结论 Itra显著抑制肺癌A549细胞增殖,与其激活AMPK诱导内质网应激性凋亡有关。
Abstract
OBJECTIVE To investigate the effects and mechanism of itraconazole (Itra) on proliferation in lung cancer A549 cell line.METHODS A549 cells were treated with indiacted dose of Itra for 24, 48 and 96h. The cell survival rate was detected by MTT and the semi-inhibitory concentration (IC50) was calculated. The cells were divided into four groups including control group, Itra group, compound C (Comp C) +Itra group, 4-PBA+ Itra group. The apoptosis was detected by AnnexinV/PI. Western blot(WB) was used to detect the expression of CHOP, GRP78, PERK, Bcl-2, Bax, AMPK, phosphorylated-AMPK(p-AMPK). RESULTS The survival rate of A549 cells was inhibited by Itra. The IC50 at 24, 48, and 96h were 62.89, 28.34 and 11.64 μmol·L-1 respectively. Compared with control group, the cell survival rate decreased, the apoptosis rate increased obviously in Itra group. Compared with group Itra, the cell survival rate decreased, the apoptosis rate increased in Comp C +Itra group and 4-PBA+ Itra group(P＜0.05). Compared with control group, the expression of CHOP, GRP78, PERK and p-AMPK increased in Itra group(P＜0.05). The expression of CHOP, GRP78, PERK and p-AMPK were lower in Comp C +Itra group and 4-PBA+ Itra group than that in control group(P＜0.05). CONCLUTION Itra induces endoplasmic reticulum stress related apoptosis via activating AMPK signaling.
关键词
伊曲康唑 /
AMPK /
内质网应激 /
细胞凋亡 /
A549细胞
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Key words
itraconazole /
AMPK /
endoplasmic reticulum stress /
apoptosis /
A549 cells
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中图分类号：
R965
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参考文献
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脚注
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基金
浙江省自然科学青年基金资助(LQ16H160019);浙江省科技厅公益性技术应用研究计划项目资助(2016C37100)
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