[1]魏攀,李杰,王莉莉,等.转染杜氏盐藻1433基因对食管鳞癌EC9706细胞Bcl2和Caspase9蛋白表达的影响*[J].郑州大学学报(医学版),2010,(01):4-7.
 WEI Pan,LI Jie,WANG Lili,et al.Expression of Bcl2 and Caspase9 proteins of EC9706 cell transfected 1433 gene of Dunaliella salina[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2010,(01):4-7.
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转染杜氏盐藻1433基因对食管鳞癌EC9706细胞Bcl2和Caspase9蛋白表达的影响*()
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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:
4-7
栏目:
食管癌研究
出版日期:
2010-01-30

文章信息/Info

Title:
Expression of Bcl2 and Caspase9 proteins of EC9706 cell transfected 1433 gene of Dunaliella salina
作者:
魏攀李杰王莉莉许培荣薛乐勋#
郑州大学生物系细胞生物学研究室郑州450001
Author(s):
WEI PanLI JieWANG LiliXU PeirongXUE Lexun
Laboratory for Cell Biology,Department of Biology,Zhengzhou University,Zhengzhou 450001
关键词:
杜氏盐藻1433基因食管鳞癌Bcl2Caspase9
Keywords:
Dunaliella salina1433 geneesophageal squamous cell cancerBcl2Caspase9
分类号:
Q789
文献标志码:
A
摘要:
探讨转染杜氏盐藻1433基因对食管鳞癌EC9709细胞Bcl2和Caspase9蛋白表达的影响。方法:将杜氏盐藻1433基因cDNA序列分别亚克隆入表达载体pEGFPC1和pcDNA3.1(+)的HindⅢ和BamHⅠ位点,构建真核表达载体pEGFPC11433和pcDNA3.1(+)1433,采用脂质体方法转染EC9706细胞,同时设立空载体对照和空白对照,荧光显微镜观察pEGFP1433融合蛋白在EC9709细胞中的定位;G418筛选稳定转染pcDNA3.1(+)1433的细胞株,采用RTPCR和Westernblotting法检测1433基因的表达;Westernblotting检测3种细胞中Bcl2和Caspase9蛋白的表达。结果:pEGFP1433融合蛋白定位于EC9706细胞的细胞核;通过G418筛选,最终获得稳定表达盐藻1433基因的转染细胞株;与转染空载体和空白对照的EC9706细胞相比,转染pcDNA3.1(+)1433的EC9706细胞中Bcl2蛋白表达量明显增高[(1.60±0.20)、(0.90±0.15)和(2.40±0.10);F=69.8897,P<0.001],而Caspase9蛋白表达量明显降低[(1.00±0.04)、(1.40±0.08)和(0.40±0.05);F=217.0667,P<0.001]。结论:杜氏盐藻1433基因可能在EC9706细胞凋亡过程中发挥了抑制作用。
Abstract:
To investigate the effect of the 1433 gene of Dunaliella salina on the expression of Bcl2 and Caspase9 proteins of esophageal squamous cell cancer cell line EC9706.Methods:The cDNA fragment of the 1433 gene of Dunaliella salina was subcloned into the HindⅢBamHⅠsites of the eukaryotic expression vector pEGFPC1 and pcDNA3.1 (+), respectively, generating recombinant expression vectors pEGFPC11433 and pcDNA3.1(+)1433.The recombinant vectors were transfected into EC9706 cells by LipofectamineTM 2000 at the same time,the vectors without 1433 genes served as controls,the location of the fusion EGFP1433 proteins was determined by fluorescent microscope and the EC9706 cells transfected with pcDNA3.1(+)1433 were selected with G418 and characterized by RTPCR and Western blotting,then the expression of the Bcl2 and Caspase9 proteins in the transfectants was monitored by Western blotting.Results:pEGFP1433 fusion proteins were localized in the nucleus of EC9706 cells. EC9706 cells stably expressing the 1433 gene of Dunaliella salina were obtained with G418 selection.The expression of Bcl2 protein significantly increased[(1.60±0.20),(0.90±0.15) and (2.40±0.10);F=69.889 7,P<0.001] while that of Caspase9 protein decreased[(1.00±0.04),(1.40±0.08) and (0.40±0.05);F=217.066 7,P<0.001] in the EC9706 cells transfected with pcDNA3.1(+)1433, compared with the control groups.Conclusion: The introduction of the 1433 gene of Dunaliella salina upregulates the expression of Bcl2 protein while downregulates that of Caspase9 protein in the EC9706 cells, suggesting its potential role in inhibiting the apoptosis of cells.

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备注/Memo

备注/Memo:
*国家自然科学基金资助项目30700014;科技部国际科技合作基金资助项目2007DFA01240#通讯作者,男,1944年生,本科,教授,研究方向:肿瘤标志物与基因工程,Email:xuelx@zzu.edu.cn
更新日期/Last Update: 2010-04-26