基于PI3K/AKT/mTOR信号通路探讨虎杖苷诱导鼻咽癌细胞凋亡的初步研究  

Study on the mechanism of polydatin inducing nasopharyngeal carcinoma apoptosis based on P13K/AKT/mTOR signaling pathway

作　　者：钟华林 李玲波[1] 覃焕桦[1] 尧振兴 ZHONG Hua lin;LI Lingbo;QIN Huanhua;YAO Zhenxing(Depatrment of Otolaryngology Head and Neck Surgery,Liuzhou Worker's Hospital(Fourth Affiliated Hospital of Guangxi Medicial University),Liuzhou,Guangxi,545000,China)

机构地区：[1]柳州市工人医院(广西医科大学第四附属医院)耳鼻咽喉头颈外科

出　　处：《中国耳鼻咽喉头颈外科》2019年第10期536-540,共5页Chinese Archives of Otolaryngology-Head and Neck Surgery

摘　　要：目的基于PI3K/AKT/m TOR信号通路探讨虎杖苷(polydatin, PYD)诱导鼻咽癌细胞凋亡的机制。方法用不同梯度浓度的PYD处理CNE-1细胞后,采用CCK-8法检测虎杖苷对CNE-1细胞增殖的抑制作用,吖啶橙/溴化乙锭(acridine orange/ethidium bromide,AO/EB)双荧光染色法显微镜观察细胞凋亡的形态学变化;流式细胞仪分细胞周期分布;q RT-PCR和Westernblot法检测细胞中PI3K、AKT、m TOR、p70S6K的m RNA和蛋白表达。结果CNE-1细胞经不同PYD(40、80、160μg/ml)干预48小时后,CNE-1细胞增殖抑制率,凋亡率呈浓度依赖性增加;G0/G1、S期细胞的百分比呈浓度依赖性降低,而G2/M期细胞的百分比呈浓度依赖性增加;CNE-1细胞中PI3K、AKT、m TOR、p70S6K的m RNA和蛋白表达明显下调,均呈浓度依赖性。结论 PYD具有抑制鼻咽癌CNE-1细胞增殖及诱导凋亡的作用,其机制可能是PI3K/AKT/m TOR信号通路受到PYD抑制,致使其下游基因蛋白表达下降。OBJECTIVE To investigate the mechanism of polydatin inducing nasopharyngeal carcinoma apoptosis based on PI3 K/AKT/mTOR signaling pathway.METHODS The growth inhibitory effect was detected with CCK-8 assay.After CNE-1 cells were treated with different concentrations of polydatin,CCK-8 assay was used to detect the inhibitory effect of polydatin on proliferation of CNE-1 cells.Annexin/propidium iodide staining was applied to detect CNE-1 cell apoptotic rate.In addition,flow cytometry was employed to analyze apoptosis and cell cycle distribution.qRT-PCR and Western blot assay were used to detect PI3 K,AKT,mTOR,and p70 S6 K mRNA and protein expression levels.RESULTS After 48 h of intervention with different PYD(40,80,160 μg/ml),the proliferation inhibition rate and apoptosis rate of CNE-1 cells increased in concentration dependence.The percentage of G0/G1 and S cells decreased in concentration dependence,while the percentage of G2/M cells increased in concentration dependence.mRNA and protein expressions of PI3 K,AKT,mTOR and p70 S6 K in CNE-1 cells were significantly down-regulated,all showing concentration dependence.CONCLUSION It indicated that polydatin could inhibit the proliferation and induce apoptosis of nasopharyngeal carcinoma CNE-1 cells,and the mechanism may be associated with inhibiting the PI3 K/AKT/mTOR signaling pathway and suppressing downstream gene expression.

关 键 词：鼻咽肿瘤 细胞凋亡 细胞增殖 虎杖苷 

分 类 号：R73[医药卫生—肿瘤]