摘要
目的研究肺炎链球菌刺激对肺腺癌细胞A549损伤的影响及作用机制。方法体外培养肺泡Ⅱ型上皮细胞A549,分为对照组和肺炎链球菌R6刺激组。R6刺激组将肺炎链球菌按照1×10~8个/ml的量加入A549细胞中进行刺激,对照组加入等体积的曲古抑素A(TSA)处理作为对照。分别处理8 h、16 h、24 h、32 h,MTT检测处理各时间点A549细胞存活情况;流式细胞仪检测R6刺激24 h后A549细胞凋亡情况;酶联免疫吸附法(ELISA)检测R6刺激24 h后炎症因子白介素6(IL-6)、IL-10含量变化;Western-blot检测R6刺激24 h后A549细胞凋亡相关蛋白及Wnt通路蛋白表达。结果 R6刺激显著抑制A549细胞存活,与对照组相比,差异具有显著性(P<0.05),R6刺激24 h对A549细胞存活抑制效果最明显。R6刺激24 h后,A549细胞凋亡率增高,与对照组相比,差异具有显著性(P<0.05)。R6刺激24 h后促炎因子IL-6含量显著增加,抗炎因子IL-10含量明显降低,与对照组相比,差异具有显著性(P<0.05)。Western-blot结果显示R6刺激后,促凋亡因子Bax、Cleaved-caspase3蛋白表达显著升高,抑凋亡因子Bcl-2表达明显下降,与对照组相比,差异具有显著性(P<0.05),Wnt通路蛋白β-catenin、p-GSK-3β表达明显升高,p-β-catenin、GSK-3β表达明显下降,与对照组相比,差异具有显著性(P<0.05)。结论肺炎链球菌刺激会对肺腺癌细胞A549损伤造成损伤,这一过程与Wnt信号通路的激活有关。
Objective To investigate the effect of Streptococcus pneumoniae on lung cancer cell A549 and its mechanisms. Methods To cultivate alveolar epithelial type Ⅱ cells in vitro,A549 cells were divided into two groups ： control group and Streptococcus pneumoniae R6 stimulation group. In the R6 stimulation group,S. pneumoniae was added to A549 cells in an amount of 1 × 10^8 cells/ml,and the control group was treated with an equal volume of TSA as a control. After treated 8 h,16 h,24 h,32 h,MTT assay was used to detect A549 cells survival. Flow cytometry was performed to detect A549 cells apoptosis after R6 stimulate 24 h. ELISA assay was used to detect A549 cells secrete inflammatory cytokines IL-6,IL-10 after R6 stimulated 24 h. Western-blot was used to detect A549 cells apoptosis protein and Wnt pathway protein expression after R6 stimulated 24 h. Results MTT assay showed that,compared with the control group,R6 stimulation could significantly inhibited A549 cell survival（P〈0. 05）,R6 stimulated A549 cell 24 h,the survival inhibitory was the most obvious. After R6 stimulation 24 h,compared with the control group,A549 cell apoptosis rate significantly increased（P〈0. 05）. Compared with the control group,the contents of IL-6 was significantly increased,but the contents of IL-10 was significantly reduced after R6 stimulated A549 cell 24 h（P〈0. 05）. Western-blot showed that,after R6 stimulated,pro-apoptotic factors Bax,Cleaved-caspase3 protein expression were significantly increased,anti-apoptotic factor Bcl-2 expression was significantly decreased（P〈0. 05）. β-catenin,p-GSK-3β expression was significantly increased,and p-β-catenin,GSK-3β expression was significantly decreased in Wnt pathway（P〈0. 05）. Conclusion Streptococcus pneumoniae stimulation can damage lung cancer cell A549,it is related to activation of the Wnt signaling pathway.
作者
吴焕良
苏雅静
吴大平
WU Huan-liang;SU Ya-jing;WU Da-ping(Department of Medical Oncology;Department of Breast Surgery,Danzhou People ＇s Hospital,Danzhou Hainan 571700,China.)
出处
《临床和实验医学杂志》
2018年第18期1933-1936,共4页
Journal of Clinical and Experimental Medicine
关键词
肺腺癌细胞
肺炎链球菌
凋亡
信号通路
Lung cancer cell
Streptococcus pneumoniae
Apoptosis
Signal pathway
