现代肿瘤医学2015年1月第23卷第2期
MODERN ONCOLOGY,Jan.2015,VOL 23,NO.02
盐酸米托葱醒通过PI3K途径诱导人鼻咽癌CNE-2细胞凋亡
主
炜，刘星言，程小广，李
涛
·153 ·
Mitoxantrone hydorchloride(MTx)induces apoptosis of human nasopharyngeal carcino
macelllineCNE-2viaPl3Kpathway Wang Wei,Liu Xingyan, Cheng Xiaoguang,Li Tao
Dongguan Research Center, Guangdong Medical Collegy, Guangdong Dongguan 523808, China.
)Objective:To investigate the effect of mitoxantrone on the prolifernation and apoptosis of human naso-
[Abstract】
pharyngeal carcinoma CNE 2 cells and its mechanism. Methods : CCK 8 method was used to observe the growth in-hibition of mitoxantrone hydrochloride on human nasopharyngeal carcinoma CNE 2 cells, The morphology of apoptot-ic 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 ICg, were 1. 9μg/ml and O. 1μg/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 aas pnpoorpeoeroosnuoaeapada aeo g aee the proliferation of CNE 2 cells and induce apoptosis,the mechanism may be associated with the reduction of the mi-
tochondrial membrane potential,and inhibition of the PI3K/Akt signaling pathway.[ Keywords ] Mitoxantrone hydrochloride ; nasopharyngeal carcinoma ; apoptosis ; PI3K
Modem Oncology 2015,23(02) :0153 0156
【摘要】
目的：探讨盐酸米托意醒对人鼻咽癌CNE-2细胞增殖和凋亡的影响及作用机制。方法：采用CC-
K-8法观察盐酸米托意醒对人鼻咽癌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；Weslermblot法检测PI3K/Akt信号相关蛋白表达。结果：盐酸米托葱醒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ICs值分别为1.9ug/ml 和0.1ug/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调亡的形态特征；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）；Westernblot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
e-3蛋白的表达。结论：盐酸米托蕙醒可显
著抑制鼻咽瘤细胞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
【关键词】盐酸米托葱醒；鼻咽瘤；凋亡;PI3K
【中图分类号】R7336*1;R739.63
【文献标识码]A
【文章编号】16724992(2015)02015304
盐酸米托醒(Mitoxantronehydorchloride，MTX)是细胞周期非特异性广谱抗癌药，它通过与DNA结合且抑制拓扑异构酶ⅡI的活性，使肿瘤细胞的周期阻滞在S和G,/M期(1
【收稿日期】【修回日期】【基金项目】
【作者单位】[作者简介】
2014 08 05 2014 08 25
东芜市医疗卫生重点项目（编号：2011105102016）；东莞市高校项目（编号：2011108102026）
广东医学院东莞科研中心，广东
东莞
523808
王炜（1981-），女，潮南衡阳人，实验师，硕士研究生，主要从事肿瘤的早期诊断与治疗研究。E-mail：wan-gei9842aliyun.com
DOI;10. 3969/j. issn. 1672 4992. 2015. 02. 03
临床上MTX广泛应用于治疗淋巴瘤、乳腺癌和急性白血病等，具有良好的疗效[2-3]，有报道称磷脂酰肌醇3-激酶/蛋白激酶B(PI3K-Akt)信号转导通路对肿瘤细胞增殖、分化和化疗敏感性起着重要的调节作用，募咽瘤中PI3K信号的
信号的调节[6]。本实验以CNE-2细胞为研究对象，观察 MTX体外诱导细胞调亡情况，并检测相关蛋白p-Akt、Akt、 Bax、Bcl-2和 Caspase-3水平的变化，以探讨MTX通过 PI3K/Akt信号通路调控凋亡的机制。
材料和方法 1
1.1材料