东南大学学报(医学版)
東南大學學報(醫學版)
동남대학학보(의학판)
JOURNAL OF SOUTHEAST UNIVERSITY(MEDICAL SCIENCE EDITION)
2015年
2期
191-195
,共5页
张倩璐%邓珊%江青山%谭俞佳%沈宝茗
張倩璐%鄧珊%江青山%譚俞佳%瀋寶茗
장천로%산산%강청산%담유가%침보명
SAA蛋白%鼻咽癌%BCL-2%Caspase-3%NF-κB
SAA蛋白%鼻嚥癌%BCL-2%Caspase-3%NF-κB
SAA단백%비인암%BCL-2%Caspase-3%NF-κB
serum amyloid protein A%nasopharyngeal carcinoma%BCL-2%Caspase-3%NF-κB
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。
目的:探討血清澱粉樣蛋白A( SAA)的超錶達和抑製錶達對BCL-2、Caspase-3及NF-κB這3種信號分子蛋白錶達的影響。方法:將課題組前期已構建成功的pcDNA3.1(+)-SAA超錶達載體和pGPU6/GFP/Neo-SAA錶達榦擾載體採用脂質體法分彆轉染鼻嚥癌CNE2細胞,建立SAA蛋白超錶達的pcDNA3.1(+)-SAA-CNE2細胞繫和榦擾SAA蛋白錶達的pGPU6/GFP/Neo-SAA-CNE2細胞繫。 pcDNA3.1(+)-SAA-CNE2細胞、CNE2細胞、pGPU6/GFP/Neo-SAA-CNE2細胞中SAA蛋白的錶達量會逐漸減少,噹SAA蛋白的錶達量改變時,通過免疫印跡法檢測BCL-2、Caspase-3及NF-κB在pcDNA3.1(+)-SAA-CNE2細胞、CNE2細胞、pGPU6/GFP/Neo-SAA-CNE2細胞中的錶達。結果:SAA蛋白的錶達在pcDNA3.1(+)-SAA-CNE2細胞繫中明顯增多(P<0.05),在pGPU6/GFP/Neo-SAA-CNE2細胞繫中顯著減少(P<0.05),由此可知, pcDNA3.1(+)-SAA超錶達載體和pGPU6/GFP/Neo-SAA錶達榦擾載體成功轉染CEN2細胞。免疫印跡檢測結果示:隨著SAA蛋白錶達的增多,BCL-2的錶達增多(P<0.05)、Caspase-3及NF-κB的錶達減少(P<0.05)。結論:SAA蛋白在人鼻嚥癌CNE2細胞中的錶達與抑製凋亡的BCL-2蛋白錶達有協同作用,對凋亡調節分子Caspase-3及NF-κB蛋白的錶達有抑製作用,可推測SAA蛋白可能有促進腫瘤生長、轉移的作用。
목적:탐토혈청정분양단백A( SAA)적초표체화억제표체대BCL-2、Caspase-3급NF-κB저3충신호분자단백표체적영향。방법:장과제조전기이구건성공적pcDNA3.1(+)-SAA초표체재체화pGPU6/GFP/Neo-SAA표체간우재체채용지질체법분별전염비인암CNE2세포,건립SAA단백초표체적pcDNA3.1(+)-SAA-CNE2세포계화간우SAA단백표체적pGPU6/GFP/Neo-SAA-CNE2세포계。 pcDNA3.1(+)-SAA-CNE2세포、CNE2세포、pGPU6/GFP/Neo-SAA-CNE2세포중SAA단백적표체량회축점감소,당SAA단백적표체량개변시,통과면역인적법검측BCL-2、Caspase-3급NF-κB재pcDNA3.1(+)-SAA-CNE2세포、CNE2세포、pGPU6/GFP/Neo-SAA-CNE2세포중적표체。결과:SAA단백적표체재pcDNA3.1(+)-SAA-CNE2세포계중명현증다(P<0.05),재pGPU6/GFP/Neo-SAA-CNE2세포계중현저감소(P<0.05),유차가지, pcDNA3.1(+)-SAA초표체재체화pGPU6/GFP/Neo-SAA표체간우재체성공전염CEN2세포。면역인적검측결과시:수착SAA단백표체적증다,BCL-2적표체증다(P<0.05)、Caspase-3급NF-κB적표체감소(P<0.05)。결론:SAA단백재인비인암CNE2세포중적표체여억제조망적BCL-2단백표체유협동작용,대조망조절분자Caspase-3급NF-κB단백적표체유억제작용,가추측SAA단백가능유촉진종류생장、전이적작용。
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.