SAA蛋白与BCL-2、Caspase-3及NF-<em>κ</em>B关系的研究-东南大学学报医学版

SAA蛋白与BCL-2、Caspase-3及NF-κB关系的研究

单位：1. 南华大学附属第一医院麻醉科, 湖南衡阳 421001;
2. 南华大学附属第一医院耳鼻咽喉科, 湖南衡阳 421001

分类号：R739.63

出版年·卷·期（页码）：2015·34·第二期（191-195）

摘要：

目的:探讨血清淀粉样蛋白A(SAA)的超表达和抑制表达对BCL-2、Caspase-3及NF-κB这3种信号分子蛋白表达的影响.方法:将课题组前期已构建成功的pcDNA3.1(+)-SAA超表达载体和pGPU6/GFP/Neo-SAA表达干扰载体采用脂质体法分别转染鼻咽癌CNE2细胞,建立SAA蛋白超表达的pcDNA3.1(+)-SAA-CNE2细胞系和干扰SAA蛋白表达的pGPU6/GFP/Neo-SAA-CNE2细胞系.pcDNA3.1(+)-SAA-CNE2细胞、CNE2细胞、pGPU6/GFP/Neo-SAA-CNE2细胞中SAA蛋白的表达量会逐渐减少,当SAA蛋白的表达量改变时,通过免疫印迹法检测BCL-2、Caspase-3及NF-κB在pcDNA3.1(+)-SAA-CNE2细胞、CNE2细胞、pGPU6/GFP/Neo-SAA-CNE2细胞中的表达.结果:SAA蛋白的表达在pcDNA3.1(+)-SAA-CNE2细胞系中明显增多(P <0.05),在pGPU6/GFP/Neo-SAA-CNE2细胞系中显著减少(P <0.05),由此可知,pcDNA3.1(+)-SAA超表达载体和pGPU6/GFP/Neo-SAA表达干扰载体成功转染CEN2细胞.免疫印迹检测结果示:随着SAA蛋白表达的增多,BCL-2的表达增多(P <0.05)、Caspase-3及NF-κB的表达减少(P <0.05).结论:SAA蛋白在人鼻咽癌CNE2细胞中的表达与抑制凋亡的BCL-2蛋白表达有协同作用,对凋亡调节分子Caspase-3及NF-κB蛋白的表达有抑制作用,可推测SAA蛋白可能有促进肿瘤生长、转移的作用.

Objective: To investigate the effect of over expression and inhibition expression of SAA protein on protein expression of BCL-2, Caspase-3 and NF-κB these three signaling molecules. Methods: Transfect the pcDNA3.1 (+)-SAA over expression vector and pGPU6/GFP/Neo-SAA expression interference vector which previously successfully constructed by the research group to nasopharyngeal carcinoma CNE2 cells respectively with liposome. Constructing pcDNA3.1(+)-SAA-CNE2 over expression cells and pGPU6/GFP/Neo-SAA-CNE2 interference cells of SAA protein. The expression of SAA protein was gradually reduce respectively in pcDNA3.1(+)-SAA-CNE2 over expression cells, CEN2 cells and pGPU6/GFP/Neo-SAA-CNE2 interference cells. Inspecting the expression of BCL-2, Caspase-3 and NF-κB in pcDNA3.1(+)-SAA-CNE2 cells and pGPU6/GFP/Neo-SAA-CNE2 cells by Western-blotting. Results: Expression of SAA protein in pcDNA3.1 (+)-SAA-CNE2 cell lines significantly increased (P <0.05), expression of SAA protein in pGPU6/GFP/Neo-SAA-CNE2 cell lines significantly decreased (P <0.05). The expression of SAA protein would decrease respectively in pcDNA3.1(+)-SAA-CNE2 over expression cells, CEN2 cells and pGPU6/GFP/Neo-SAA-CNE2 interference cells, the difference was statistically significant. Western-blotting showed that with the increase expression of SAA protein, expression of BCL-2 increased (P <0.05), but expression of Caspase-3 and NF-κB decreased(P <0.05). Conclusion: For the expression of SAA protein in human nasopharyngeal carcinoma cells CNE2, there is a positive correlation with expression of BCL-2 which restraining of apoptosis, a negative correlation with expression of Caspase-3 and NF-κB which regulating apoptosis. We can make a speculation that SAA protein may have the effect of promoting tumor growth and metastasis.

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