[1]亚国伟,张威,陈玉,等.PTEN反义寡核苷酸对EC9706细胞增殖、凋亡和PTEN、mTOR表达的影响*[J].郑州大学学报(医学版),2010,(01):16-20.
 YA Guowei,ZHANG Wei,CHEN Yu,et al.Changes of proliferation, apoptosis and expression of mTOR and PTEN in EC9706 cells after gene PTEN inhibition by antisense oligonucleotides[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2010,(01):16-20.
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PTEN反义寡核苷酸对EC9706细胞增殖、凋亡和PTEN、mTOR表达的影响*()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2010年01期
页码:
16-20
栏目:
食管癌研究
出版日期:
2010-01-30

文章信息/Info

Title:
Changes of proliferation, apoptosis and expression of mTOR and PTEN in EC9706 cells after gene PTEN inhibition by antisense oligonucleotides
作者:
亚国伟张威陈玉陈奎生#
郑州大学第一附属医院病理科;河南省肿瘤病理重点实验室郑州450052
Author(s):
YA GuoweiZHANG Wei CHEN YuCHEN Kuisheng
Department of Pathology,the First Affiliated Hospital,Zhengzhou University;Henan Key Laboratory of Tumor Pathology,Zhengzhou 450052
关键词:
PTEN反义寡核苷酸mTOR增殖凋亡EC9706细胞
Keywords:
PTENantisense oligonucleotidemTORproliferationapoptosisEC9706 cell
分类号:
R735.1
文献标志码:
A
摘要:
探讨PTEN反义寡核苷酸(ASODN)对食管癌EC9706细胞增殖、凋亡和PTEN、mTOR表达的影响。方法:利用阳离子脂质体介导PTEN的ASODN和无义寡核苷酸(NODN)转染食管癌EC9706细胞,终浓度为3、6和12μmol/L的ASODN为实验组,终浓度12μmol/L的NODN为NODN对照组,不进行转染的为空白对照组。运用MTT法、TUNEL检测空白对照组、NODN对照组及3种浓度的ASODN作用24、48及72h后食管癌EC9706细胞增殖和凋亡情况;采用免疫细胞化学技术及原位杂交方法检测各组EC9706细胞中PTEN蛋白、mTOR蛋白和mRNA的表达。结果:①随PTENASODN转染浓度(F3μmol/L=784.774,F6μmol/L=242.884,F12μmol/L=412.105,P均<0.001)和时间(F24h=60.676,F48h=57.703,F72h=51.841,P<0.001)的增加,食管癌EC9706细胞的增殖增强。②随PTENASODN转染浓度(F3μmol/L=36.655,F6μmol/L=47.594,F12μmol/L=85.264,P均<0.001)和时间(F24h=4.860,F48h=4.901,F72h=8.153,P<0.001)的增加,食管癌EC9706细胞的凋亡下降。③随PTENASODN转染浓度(F3μmol/L=45.634,F6μmol/L=66.399,F12μmol/L=72.763,P均<0.001)和时间(F24h=4.065,F48h=3.985,F72h=5.446,P<0.001)的增加,PTEN的蛋白表达降低;mRNA的表达也降低(F3μmol/L=23.357,F6μmol/L=43.272,F12μmol/L=51.402,P均<0.001;F24h=3.933,F48h=4.084,F72h=4.528,P<0.001)。④mTOR蛋白(F3μmol/L=19.422,F6μmol/L=30.000,F12μmol/L=62.168,P<0.001;F24h=5.340,F48h=9.150,F72h=21.056,P<0.001)和mRNA(F3μmol/L=19.414,F6μmol/L=46.571,F12μmol/L=68.634,P<0.001;F24h=4.815,F48h=12.165,F72h=7.858,P<0.001)的表达随PTENASODN转染浓度和时间的增加而增强。⑤上述各指标中均以12μmol/LASODN作用72h的效应最强(P均<0.05)。结论:PTENASODN促进食管癌EC9706细胞增殖、抑制凋亡,下调PTEN蛋白和mRNA的表达,上调mTOR蛋白和mRNA的表达。
Abstract:
To observe the changes of proliferation,apoptosis and expression of PTEN and mTOR in EC9706 cells after the gene PTEN being inhibited by antisense oligonucleotides(ASODN).Methods:The ASODN of PTEN at 3,6,and 12 μmol/L was transfected into EC9706 cells by the cationic liposome for 24,48,and 72 hours, nonsense oligonudeotide at 12 μmol/L was transfected as unrelated control,and EC9706 cells not being transfected as normal control.The MTT and TUNEL methods were used to examine the proliferation and apoptosis of EC9706 cells respectively.The immunocytochemistry was used to determine the expressions of protein PTEN and mTOR. The in situ hybridization was used to determine the mRNA expressions of PTEN and mTOR.Results: The proliferation of esophageal cancer EC9706 cells was strengthened(F3 μmol/L=784.774,F6 μmol/L=242.884,F12 μmol/L=412.105,P<0.001;F24 h=60.676,F48 h=57.703,F72 h=51.841,P<0.001),but the apoptotic rate was descendent(F3 μmol/L=36.655,F6 μmol/L=47.594,F12 μmol/L=85.264,P<0.001;F24 h=4.860,F48 h=4.901,F72 h=8.153,P<0.001),and expressions of PTEN protein(F3 μmol/L=45.634,F6 μmol/L=66.399,F12 μmol/L=72.763,P<0.001;F24 h=5.065,F48 h=3.985,F72 h=5.446,P<0.001) and mRNA(F3 μmol/L=23.357,F6 μmol/L=43.272,F12 μmol/L=51.402,P<0.001;F24 h=3.933,F48 h=4.084,F72 h=4.528,P<0.001) were reduced,while expressions of mTOR protein(F3 μmol/L=19.422,F6 μmol/L=30.000,F12 μmol/L=62.168,P<0.001;F24 h=5.340,F48 h=9.150,F72 h=21.056,P<0.001) and mRNA(F3 μmol/L=19.414,F6 μmol/L=46.571,F12 μmol/L=68.634,P<0.001;F24 h=4.815,F48 h=12.165,F72 h=7.858,P<0.001) were enhanced in EC9706 cells in time and dosedependent manner.The dose of 12 μmol/L for 72 hours was exhibited the most strongest effect(P<0.05).Conclusion:The ASODN of PTEN could enhance the proliferation of esophageal cancer EC9706 cells and the expression of mTOR,but could downregulate the apoptotic rate and the expression of PTEN protein and mRNA.

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备注/Memo

备注/Memo:
*河南省杰出青年科学基金资助项目074100510009;河南省科技攻关计划基金资助项目082102310011;河南省基础与前沿技术研究计划基金资助项目092300410016;郑州市科技创新团队基金资助项目 #通讯作者,男,1964年生,博士,教授,研究方向:肿瘤病理
更新日期/Last Update: 2010-04-26