Beclin1对喉鳞癌细胞Hep-2紫杉醇敏感性影响的体外研究

邓晓聪; 杨新明; 李仕晟

doi:10.13201/j.issn.1001-1781.2015.02.016

Beclin1对喉鳞癌细胞Hep-2紫杉醇敏感性影响的体外研究

- 1 海口市人民医院耳鼻咽喉科(海口, 570311);
- 2 中南大学湘雅二医院耳鼻咽喉科

详细信息

通讯作者: 李仕晟,E-mail:hksyydxc@163.com

中图分类号: R739.65

收稿日期: 2014-08-28

The vitro research of effects of Beclin1 on paclitaxel-sensitivity in laryngeal carcinoma cell Hep-2

- 1 Department of Otorhinolaryngology, People's Hospital of Haikou, Haikou, 570311, China;
- 2 Department of Otorhinolaryngology, Xiangya No. 2 Hospital of Central South University

More Information

Corresponding author: LI Shisheng, E-mail: hksyydxc@163.com

Received Date: 28 August 2014

摘要

摘要: 目的:通过检测不同Beclin1表达水平对喉癌细胞紫杉醇敏感性的影响。方法:本研究利用以喉癌细胞株Hep-2、稳定转染pcDNA3.1-Beclin1质粒的喉癌细胞株Hep-2-Beclin1、稳定转染pcDNA3.1质粒载体的喉癌细胞株Hep-2-pcDNA3.1为研究对象,对3组细胞施加不同浓度的化疗药物紫杉醇(1、2、5、10、20 μg/L)干预24 h,利用MTT法及流式细胞术检测紫杉醇对3组细胞增殖和凋亡的影响。利用Western blot检测3组细胞Akt和p-Akt蛋白表达水平。结果:不同浓度的紫杉醇处理后的3组喉癌细胞与药物终浓度正相关地出现生长抑制率提高。当紫杉醇浓度大于5 μg/L时,紫杉醇对Hep-2-Beclin1的生长抑制率高于对另两株细胞的生长抑制率。以终浓度为10、20 μg/L的紫杉醇处理3株喉癌细胞24 h后,流式细胞术检测结果显示:各组细胞20 μg/L紫杉醇处理后细胞的凋亡率均高于10 μg/L紫杉醇处理后细胞的凋亡率(P<0.05)。紫杉醇处理后Hep-2-Beclin1组细胞凋亡率均高于Hep-2组和Hep-2-pcDNA3.1组(P<0.05)。 Western blot结果显示:Hep-2、Hep-2-pcDNA3.1、Hep-2-Beclin1细胞Akt相对蛋白表达量分别为:1.24±0.03、1.25±0.05、1.27±0.09,3组细胞间Akt相对蛋白表达量差异无统计学意义(P>0.05);Hep-2、Hep-2-pcDNA3.1、Hep-2-Beclin1细胞p-Akt即活化型Akt相对蛋白表达量分别为:0.98±0.09、1.03±0.04、0.54±0.03,Hep-2-Beclin1细胞p-Akt相对蛋白表达量低于Hep-2、Hep-2-pcDNA3.1组细胞(P<0.05)。结论:Beclin1可能通过抑制PI3K/Akt信号通路的活化上调喉癌细胞对紫杉醇的敏感性。
- Beclin1 /
- 喉癌 /
- 自噬 /
- 紫杉醇
Abstract: Objective: Background: We detect the effects of Beclin1 on paclitaxel-sensitivity in laryngeal carcinoma cell.Method: This study used Hep-2, Hep-2-pcDNA3.1, Hep-2-Beclin1 as invitro model. The effect of paclitaxel on the proliferation and cell apoptosis of laryngeal cancer cell lines was evaluated by MTT assay and flow cytometry. The protein expression level of Akt and p-Akt was detected by Western blot.Result: After treated by paclitaxel, the inhibition rate was significantly higher in Hep-2-Beclin cells than in Hep-2-pcDNA3.1 cells and Hep-2 cells (P<0.05). After dealing with 10 μg/L paclitaxel, the apoptosis rate in Hep-2, Hep-2-pcDNA3.1, Hep-2-Beclin1 were (23.75±3.77)%, (21.25±4.92)%, (32.50±5.97)%, respectively. After dealing with 20μg/L paclitaxel, the apoptosis rate in Hep-2, Hep-2-pcDNA3.1, Hep-2-Beclin1 were (38.75±4.79)%, (38.75±6.55)%, (50.00±7.26)%, respectively. Paclitaxel-induced apoptosis was higher in Hep-2-Beclin cells than in Hep-2-pcDNA3.1 cells and Hep-2 cells (P<0.05). The result of western blot showed that the protein expression level of p-Akt in Hep-2-Beclin cells was lower than in Hep-2-pcDNA3.1 cells and Hep-2 cells (P<0.05) and the protein expression level of Akt was similar in three cell lines (P>0.05).Conclusion: Beclin1 enhances paclitaxel-sensitivity by inhibition of PI3K/Akt pathway.
- Beclin1 /
- laryngeal squamous cell carcinoma /
- autophagy /
- paclitaxe

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