下调FoxM1通过激活JNK/线粒体通路增加人鼻咽癌细胞对紫杉醇的敏感性

雷越; 万婕; 李丹丹; 叶琳; 刘亚男; 祝琳; 陈鸿雁

doi:10.16016/j.1000-5404.201711248

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下调FoxM1通过激活JNK/线粒体通路增加人鼻咽癌细胞对紫杉醇的敏感性

基础医学 | 更新时间：2026-01-09

- 下调FoxM1通过激活JNK/线粒体通路增加人鼻咽癌细胞对紫杉醇的敏感性
- Down-regulation of FoxM1 enhances sensitivity of human nasopharyngeal carcinoma cells to paclitaxel via activation of JNK/mitochondrial pathway
- 第三军医大学学报 2018年40卷第9期页码：780-787
- 作者机构：
  
  1.400016 重庆, 重庆医科大学附属第一医院:耳鼻喉头颈外科
  2.400016 重庆, 重庆医科大学附属第一医院:胸心外科
  3.332000 江西九江, 江西省九江市第一人民医院耳鼻喉头颈外科
  4.646000 四川泸州, 西南医科大学附属医院耳鼻咽喉-头颈外科
- 作者简介：
  
  [陈鸿雁, E-mail:chenhongyan1963@hotmail.com CHEN Hongyan, E-mail:chenhongyan1963@hotmail.com
- 基金信息：
  
  重庆市渝中区科技计划项目(20120203)
- DOI：10.16016/j.1000-5404.201711248
  中图分类号： R73-362;R739.63;R979.19
- 收稿：2017-11-29，
  
  修回：2018-01-29，
  
  纸质出版：2018-05-15
- 稿件说明：
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雷越, 万婕, 李丹丹, 等. 下调FoxM1通过激活JNK/线粒体通路增加人鼻咽癌细胞对紫杉醇的敏感性[J]. 第三军医大学学报, 2018,40(9):780-787.

Yue LEI, Jie WAN, Dandan LI, et al. Down-regulation of FoxM1 enhances sensitivity of human nasopharyngeal carcinoma cells to paclitaxel via activation of JNK/mitochondrial pathway[J]. Journal of Third Military Medical University, 2018, 40(9): 780-787.
雷越, 万婕, 李丹丹, 等. 下调FoxM1通过激活JNK/线粒体通路增加人鼻咽癌细胞对紫杉醇的敏感性[J]. 第三军医大学学报, 2018,40(9):780-787. DOI： 10.16016/j.1000-5404.201711248.

Yue LEI, Jie WAN, Dandan LI, et al. Down-regulation of FoxM1 enhances sensitivity of human nasopharyngeal carcinoma cells to paclitaxel via activation of JNK/mitochondrial pathway[J]. Journal of Third Military Medical University, 2018, 40(9): 780-787. DOI： 10.16016/j.1000-5404.201711248.

摘要

目的

探讨特异性小干扰RNA(small interfering RNA

siRNA)沉默转录因子叉头框M1(FoxM1)对人鼻咽癌HONE-1细胞增殖、凋亡、紫杉醇敏感性的影响及可能的分子机制。

方法

采用siRNA靶向沉默鼻咽癌HONE-1细胞FoxM1表达并通过RT-PCR、实时定量PCR、Western blot检测转染后FoxM1的表达水平。siRNA转染后细胞的增殖和对紫杉醇的敏感性采用MTT法检测

流式细胞仪检测细胞周期分布

Annexin V-FITC/PI双染法检测细胞凋亡率。Western blot检测Ki67、CyclinE1、多药耐药基因1(multidrug resistance 1

MDR1)及c-Jun氨基末端激酶(c-Jun N-terminal kinase

JNK)、p-JNK、c-Jun、p-c-Jun、凋亡相关蛋白Bax、Bcl-2的表达水平。

结果

转染后FoxM1 mRNA及蛋白的表达均被下调(

0.01)。siRNA转染组细胞增殖速率减缓(

0.05)

Ki67表达降低(

0.05)。siRNA转染组G

期细胞比例增加(

0.01)

S期比例减低(

0.05)

Cyclin E1低表达(

0.01)。同时

siRNA+紫杉醇组凋亡率明显高于阴性及空白对照+紫杉醇组(

0.01)。siRNA转染组细胞MDR1水平下降(

0.05)

对紫杉醇的敏感性较两对照组明显增强(

0.01)。siRNA+紫杉醇组与阴性对照+紫杉醇组相比p-JNK1、p-c-Jun、Bax表达上调(

0.01)

Bcl-2表达下调(

0.01)。

结论

特异性siRNA沉默FoxM1表达可以影响JNK/线粒体通路活性

抑制鼻咽癌HONE-1细胞增殖

促进其凋亡

提高其对紫杉醇的敏感性。

Abstract

Objective

To determine the effects of down-regulating Forkhead box M1 gene (FoxM1) on the proliferation

apoptosis and sensitivity to paclitaxel in human nasopharyngeal carcinoma cell line HONE-1

and investigate the possible molecular mechanism.

Methods

A sequence-specific small interfering RNA (siRNA) targeting FoxM1 was used to transfect HONE-1 cells

then the expression of FoxM1 was detected by quantitative reverse transcription-PCR (qRT-PCR)

RT-PCR and Western blotting.The proliferation and chemosensitivity to paclitaxel cells were detected by MTT assay.Flow cytometry was adopted to measure the cell cycle distribution of transfected HONE-1 cells.The apoptotic rates were examined by Annexin V-FITC/PI staining assay.And the expression levels of Ki67

Cyclin E1

multidrug resistance 1 (MDR1)

c-Jun N-terminal kinase (JNK)

p-JNK

c-Jun

p-c-Jun

and apoptosis-related proteins Bax and Bcl-2 were detected by Western blotting.

Results

FoxM1-siRNA transfection decreased the expression level of FoxM1 gene in HONE-1 cells obviously (

0.01).Compared with the control cells

the transfected HONE-1 cells had markedly decreased proliferation rate (

0.05)

much lower Ki67 expression (

0.05)

increased percentage of the cells in G

phase (

0.01) while less cells in S phase (

0.05)

and down-regulated expression of Cyclin E1 (

0.01).Meanwhile the apoptotic rate was significantly higher in the siRNA/paclitaxel cells than negative and blank control/paclitaxel cells (

0.01).What's more

the transfection obviously decreased the MDR1 level (

0.05)

and significantly enhanced the sensitivity of HONE-1 cells to paclitaxel (

0.01)

when compared with the control groups.In siRNA/paclitaxel group

the expression levels of p-JNK1

p-c-Jun and Bax were much higher than the negative control/paclitaxel group (all

0.01)

whereas that of Bcl-2 was down-regulated (

0.01).

Conclusion

The siRNA targeting FoxM1 in HONE-1 cells could effectively regulate the activity of JNK/mitochondrial pathway

inhibit the proliferation

promote the apoptosis

and ultimately enhance the sensitivity to paclitaxel.

关键词

Keywords

references

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