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中国癌症防治杂志

中国癌症防治杂志 ›› 2011, Vol. 3 ›› Issue (4): 284-287.doi: 10.3969/j.issn.1674-5671.2011.04.04

• 基础研究 • 上一篇    下一篇

低氧条件下喉癌Hep-2细胞Stat3和HIF-1α表达关系的研究

路秀英, 李晓明   

  1. 石家庄白求恩国际和平医院耳鼻咽喉-头颈外科；全军耳鼻咽喉头颈外科中心
  • 出版日期:2011-12-25 发布日期:2012-01-20
  • 通讯作者: 李晓明 E-mail:xmlmo@126.com
  • 基金资助:

    全军医药卫生十一五科技攻关项目（200606M076）

The relationship between the expressions Stat3 and HIF-1α in Hep-2 cells under hypoxia

LU Xiu-Ying, LI Xiao-Ming   

  • Online:2011-12-25 Published:2012-01-20

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摘要： 目的 探讨低氧条件下喉癌Hep-2细胞磷酸化信号转导子和转录激活子3（phosphorylation signal transducer and activator of transcription 3，p-Stat3）及低氧诱导因子1α (hypoxia inducible factor 1α，HIF-1α)、VEGF的表达及Stat3 反义寡核苷酸（antisense oligodenucleotides，AS-ODN)对其表达的影响。方法 应用流式细胞术检测常氧和低氧不同时间Hep-2细胞p-Stat3、HIF-1α、VEGF蛋白表达，检测Stat3 AS-ODN转染Hep-2细胞24h，再于低氧条件下培养6h后Hep-2细胞p-Stat3、HIF-1α、VEGF蛋白表达。应用MTT法检测不同浓度（100、200nmol/L）Stat3 AS-ODN、Stat3正义寡核苷酸（sense oligodenucleotides，S-ODN）、脂质体作用于Hep-2细胞后的细胞存活率。结果 与常氧培养相比较，低氧培养条件下Hep-2细胞p-Stat3表达的荧光指数（fluorescent index，FI）在3h即有增长，6h后稳定；HIF-1α及VEGF的FI值6h和12h后增长，p-Stat3表达的变化与HIF-1α变化呈明显正相关性（r=0.691，P＜0.01）。低氧条件下Stat3 AS-ODN转染Hep-2细胞后抑制p-Stat3表达，HIF-1α、VEGF的蛋白表达FI值也随之下降，Pearson相关分析显示其变化呈正相关性(r=0.912，P＜0.01)。Stat3 AS-ODN转染Hep-2细胞后MTT细胞存活率随Stat3 AS-ODN浓度（100、200nmol/L）增加而下降，与Stat3 S-ODN对照组和脂质体对照组比较差异均有统计学意义（P均＜0.01）。结论喉癌Hep-2细胞中存在由低氧激活的Stat3信号转导，p-Stat3于HIF-1α及VEGF表达之前出现增长，阻断Stat3后低氧条件下HIF-1α表达降低，提示Stat3调节 HIF-1α表达。

关键词: Stat3, 低氧, 反义寡核苷酸, HIF-1α, 喉癌

Abstract: Objective To investigate the expressions of p-Stat3，HIF-1α and VEGF in Hep-2 cells under hypoxia，and the effect of Stat3 antisense oligonucleotides (AS-ODN) on these expressions.Methods Expressions of p-Stat3，HIF-1α and VEGF of Hep-2 cells that were with or without Stat3 AS-ODN transfection，at hypoxic and normoxic conditions，were detected by flow cytometry.The survival rate of Hep-2 cells transfected with Stat3 AS-ODN and control groups were analyzed by MTT assay.Results The FI of p-Stat3 was up-regulated in 3h-hypoxia group，and it was stable after 6h at hypoxic condition.The FI of HIF-1α and VEGF increased after 6h and 12h at hypoxic condition. There was positive correlation between the expression of p-Stat3 and HIF-1α (r=0.691，P＜0.01).Compared to Stat3 S-ODN group and   lipofectamine  control group，the expressions of p-Stat3，HIF-1α and VEGF in Stat3 AS-ODN group at hypoxic condition were significantly decreased (P＜0.01 ). Pearson correlation analysis showed the significant positive correlation between the expressions of p-Stat3 and HIF-1α (r=0.912，P＜0.01).The survival rate of Hep-2 cell   reduced with the concentration augmentation of Stat3 AS-ODN (100nmol/L and 200nmol/L)，which were significantly different from the Stat3 S-ODN group and  HIF- 1α  and VEGFlipofectamine control group (P＜0.01).Conclusions Hypoxia can induce Stat3 activation in Hep-2 cells.The expression of p-Stat3 is earlier than that of  HIF-1α and VEGF.Blocking Stat3 can decrease the expression of HIF-1α under hypoxia，indicating Stat3 might modulate HIF-1α expression in Hep-2 cells at the hypoxic condition.

Key words: Stat3, Hypoxia, Antisense oligonucleotides, HIF-1α, Laryngeal carcinoma