盐酸米托蒽醌通过PI3K途径诱导人鼻咽癌CNE-2细胞凋亡

Mitoxantrone hydorchloride( MTX) induces apoptosis of human nasopharyngeal carcinoma cell line CNE- 2 via PI3K pathway

目的:探讨盐酸米托蒽醌对人鼻咽癌CNE-2细胞增殖和凋亡的影响及作用机制。方法:采用CCK-8法观察盐酸米托蒽醌对人鼻咽癌CNE-2细胞生长的影响;AO/EB染色法观察凋亡细胞形态;流式细胞术检测细胞内线粒体膜电位;Western blot法检测PI3K/Akt信号相关蛋白表达。结果:盐酸米托蒽醌可明显抑制鼻咽癌细胞CNE-2的增殖,且呈时间和剂量依赖性。24h和48h半数抑制浓度IC50值分别为1.9μg/ml和0.1μg/ml。凋亡染色显示细胞在药物作用48h后出现细胞核染质浓缩、边缘化以及凋亡小体形成等细胞凋亡的形态特征;流式检测到药物处理后的细胞线粒体膜电位明显降低(P<0.05);Western blot显示药物呈剂量依赖地抑制p-Akt、Bcl-2蛋白的表达,上调Bax和Caspase-3蛋白的表达。结论:盐酸米托蒽醌可显著抑制鼻咽癌细胞CNE-2的增殖并诱导其凋亡,其机制可能与其降低细胞线粒体膜电位,并抑制PI3K/Akt信号通路有关。

Objective：To investigate the effect of mitoxantrone on the proliferation and apoptosis of human nasopharyngeal carcinoma CNE-2 cells and its mechanism.Methods：CCK-8 method was used to observe the growth inhibition of mitoxantrone hydrochloride on human nasopharyngeal carcinoma CNE-2 cells.The morphology of apoptotic cells was observed by AO/EB staining.Mitochondrial membrane potential of CNE-2 cells was measured by flow cytometry.The expression of PI3K/Akt signal protein was test by Western blot.Results：Mitoxantrone hydrochloride can obviously inhibit CNE-2 proliferation in a dose-dependent manner,24h and 48h IC50 were 1.9μ g/ml and 0.1 μg/ml.Apoptosis staining cells appeared morphological characteristics of nucleus condensation,marginalization and apoptotic body cell apoptosis after treated with various concentration MTX for 48h.The mitochondrial membrane potential of mitoxantrone treated cells were significantly lower than those in the blank group （P ＜ 0.05）.Mitoxantrone hydrochloride downregulated the expression of p-Akt,Bcl-2 protein and upregulated the expression of Bax and Caspase-3 protein in a dose dependent manner.Conclusion：Mitoxantrone hydrochloride could significantly inhibit the proliferation of CNE-2 cells and induce apoptosis,the mechanism may be associated with the reduction of the mitochondrial membrane potential,and inhibition of the PI3K/Akt signaling pathway.