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转染IL-27基因对人肺癌A549细胞增殖及STAT3表达的影响
作者:黄蓉飞1  黄娜2  樊元春1  王晓燕1  余咸静1  张君莉1  熊怡凡1 
单位:1. 成都医学院第一附属医院病理科, 四川 成都 610500;
2. 成都医学院第一附属医院呼吸内科, 四川 成都 610500
关键词:人肺癌A549细胞 转染 白细胞介素27 增殖 信号转导与转录激活因子3 
分类号:R734.2
出版年·卷·期(页码):2019·38·第十期(1191-1197)
摘要:

目的:探讨转染白细胞介素27(IL-27)基因对人肺癌A549细胞增殖及信号转导与转录激活因子3(STAT3)表达的影响。方法:培养A549细胞并分为A549细胞组(未转染细胞)、A549-HisB细胞组(转染pcDNA3.1-HisB空质粒的A549细胞)、A549-IL27细胞组(转染pcDNA3.1-HisB-IL-27质粒的A549细胞)。RT-PCR和ELISA法检测IL-27转染结果,CCK-8法检测转染IL-27基因对A549细胞增殖的影响,流式细胞仪检测转染IL-27基因对A549细胞周期及凋亡的影响,蛋白质印迹法和qRT-PCR检测A549细胞中STAT3蛋白和mRNA表达情况。结果:在A549-IL27细胞中扩增到IL-27p28基因和IL-27EB13基因条带,而A549细胞和A549-HisB细胞中未扩增到;转染后细胞培养4 h,在A549-IL27细胞培养液中可检测到IL-27表达,而A549细胞和A549-HisB细胞培养液中检测不到;转染后细胞培养24、36、48、60 h时,A549-IL27细胞增殖明显低于A549细胞和A549-HisB细胞(P<0.05);与A549细胞和A549-HisB细胞相比,A549-IL27细胞G0/G1期比例明显增多,G2/M期比例明显减少,细胞凋亡率明显升高(P<0.05);A549-IL27细胞中STAT3蛋白和mRNA表达水平均低于A549细胞和A549-HisB细胞(P<0.05);A549-IL27细胞中B淋巴细胞瘤-2(Bcl-2)蛋白表达降低,细胞淋巴瘤/白血病-2相关X蛋白(Bax)、含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)蛋白表达升高,与A549细胞和A549-HisB细胞比较差异有统计学意义(P<0.05)。结论:转染IL-27基因能抑制人肺癌A549细胞增殖,促进细胞凋亡,可能与抑制STAT3表达,阻断其信号传导有关。

Objective:To investigate the effects of interleukin 27(IL-27) gene transfection on proliferation of human lung cancer A549 cells and expression of signal transducer and activator of transcription 3(STAT3).Methods:A549 cells were cultured and divided into A549 cell group(non-transfected cells), A549-HisB cell group(A549 cells transfected with pcDNA3.1-HisB empty plasmid), A549-IL27 cell group(A549 cells transfected with pcDNA3.1-HisB-IL-27 plasmid). RT-PCR and ELISA were used to detect the transfection results of IL-27, CCK-8 assay was used to detect the effect of IL-27 gene transfection on the proliferation of A549 cells,and the effects of transfection of IL-27 gene on cell cycle and apoptosis of A549 cells were determined by flow cytometry. Western blot and qRT-PCR were used to detect the expressions of STAT3 protein and mRNA in A549 cells.Results:A549-IL27 cells were amplified into IL-27p28 gene and IL-27EB13 gene band,while A549 cells and A549-HisB cells were not amplified. After 4 hours' culture of transfected cell, the expression of IL-27 could be detected in A549-IL27 cell culture medium, but it couldn't be detected in A549 cell and A549-HisB cell culture medium. After 24, 36, 48 and 60 hours' culture of transfected cells, the proliferation of A549-IL27 cells decreased significantly, which was lower than that of A549 cells and A549-HisB cells(P<0.05). Compared with A549 cells and A549-HisB cells, the proportion of G0/G1 phase in A549-IL27 cells increased significantly, the proportion of G2/M phase decreased significantly, and the apoptotic rate increased significantly(P<0.05). The expression levels of STAT3 protein and mRNA in A549-IL27 cells decreased significantly, which were lower than those in A549 cells and A549-HisB cells(P<0.05). The expression of B-cell lymphoma-2(Bcl-2) protein in A549-IL27 cells decreased and the expressions of B cell lymphoma/leukmia-2 associated X protein(Bax) and cysteinyl aspartate specific proteinase 3(Caspase-3) proteins increased, which were significantly different from those in A549 cells and A549-HisB cells(P<0.05). Conclusion:Transfection of IL-27 gene can inhibit the proliferation and promote apoptosis of human lung cancer A549 cells, which may be related to inhibiting STAT3 expression and blocking its signal transduction.

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