Fenofibrate induces cycle arrest in lung adenocarcinoma cells by down-regulating Cyclin D1 expression
王桂平 1李智斌 1周伟平 1张彦焘 1刘新艳
扫码查看
点击上方二维码区域，可以放大扫码查看
作者信息
1. 广州卫生职业技术学院药学院，广州 510180
折叠
摘要
目的 探讨非诺贝特（FEN）对肺腺癌A549细胞的增殖、细胞周期影响及其相关分子机制，为FEN治疗肺腺癌提供理论依据。 方法 实验设对照组[二甲基亚砜（DMSO）]和不同浓度FEN组（100、200、400 μmol/L），采用CCK-8法检测FEN对A549细胞增殖抑制率。将A549细胞分为对照组（DMSO）和100、200 μmol/L FEN组，通过流式细胞仪检测细胞周期；应用生物信息学技术揭示FEN抗肺腺癌分子机制，并筛选出治疗肺腺癌的核心靶点分子；应用实时荧光定量（qRT）-PCR及免疫印迹法验证FEN对核心靶点细胞周期蛋白D1（CCND1）基因及蛋白表达的影响。 结果 与对照组比较，随着FEN浓度增加，A549细胞增殖抑制率也逐渐增高（均P<0.05），且各FEN组增殖抑制率也随作用时间延长逐渐增高（均P<0.05）。FEN可诱导A549细胞产生G1期阻滞，100、200 μmol/L FEN组G1期细胞比率较对照组明显升高（均P<0.05）。生物信息学预测显示，FEN可能主要通过激动过氧化物酶增殖激活受体γ（PPARγ）和PPARα，调控CCND1等31个核心靶分子表达而发挥抗肺腺癌作用，而FEN对A549细胞G1期阻滞可能主要与CCND1的表达下调相关。qRT-PCR及免疫印迹验证实验也表明，与对照组比较，FEN可明显抑制CCND1基因及蛋白表达（均P<0.05）。 结论 FEN通过抑制CCND1分子表达而抑制A549细胞增殖，并将细胞周期阻滞在G1期。 Objective To investigate the effects of Fenofibrate (FEN) on proliferation and cell cycle of lung adenocarcinoma cell line A549 and its related molecular mechanism, so as to provide the theoretical rationale for FEN as a treatment of lung adenocarcinoma. Methods A control group (dimethyl sulfoxide, DMSO) and FEN groups at different doses (100, 200 and 400 μmol/L) were included in the study. The inhibitory rate of FEN on A549 cell proliferation was examined by CCK-8 assay. A549 cells were divided into the control group (DMSO) and 100, 200 μmol/L FEN groups, and the cell cycle was measured by flow cytometry. Bioinformatic technique was used to determine the molecular mechanism of FEN against lung adenocarcinoma, and the core target molecules for the treatment of lung adenocarcinoma were screened out. Real-time fluorescence quantitative (qRT) -PCR and Western blotting were used to verify the effect of FEN on the gene and protein expression of the core target cyclin D1 (CCND1) . Results Compared with the control group, the inhibition rate of A549 cell proliferation gradually increased along with higher concentrations of FEN (all P<0.05), and the inhibition rate of A549 cell proliferation in the FEN groups gradually increased with the time (allP<0.05). FEN could induce cell cycle arrest at G1 phase in A549 cells, and the proportions of G1 phase cells in 100 and 200μmol/L FEN groups were significantly elevated compared with that in the control group (allP<0.05). Bioinformatics prediction showed that FEN may play the role against lung adenocarcinoma mainly by activating PPARγ and PPARα receptors, and regulating the expressions of 31 core target molecules such as CCND1 the cell cycle arrest of A549 cells at G1 phase by FEN may be mainly related to down-regulation of CCND1 expression. qRT-PCR and Western blotting verified that FEN could significantly inhibit the gene and protein expression of CCND1 compared with that in the control group (all P<0.05) . Conclusion Fenofibrate may inhibit the proliferation of A549 cells by down-regulating CCND1 expression, and thereby arrest the cell cycle in G1 phase.
Abstract
Objective To investigate the effects of Fenofibrate (FEN) on proliferation and cell cycle of lung adenocarcinoma cell line A549 and its related molecular mechanism, so as to provide the theoretical rationale for FEN as a treatment of lung adenocarcinoma. Methods A control group (dimethyl sulfoxide, DMSO) and FEN groups at different doses (100, 200 and 400 μmol/L) were included in the study. The inhibitory rate of FEN on A549 cell proliferation was examined by CCK-8 assay. A549 cells were divided into the control group (DMSO) and 100, 200 μmol/L FEN groups, and the cell cycle was measured by flow cytometry. Bioinformatic technique was used to determine the molecular mechanism of FEN against lung adenocarcinoma, and the core target molecules for the treatment of lung adenocarcinoma were screened out. Real-time fluorescence quantitative (qRT) -PCR and Western blotting were used to verify the effect of FEN on the gene and protein expression of the core target cyclin D1 (CCND1) . Results Compared with the control group, the inhibition rate of A549 cell proliferation gradually increased along with higher concentrations of FEN (all P<0.05), and the inhibition rate of A549 cell proliferation in the FEN groups gradually increased with the time (allP<0.05). FEN could induce cell cycle arrest at G1 phase in A549 cells, and the proportions of G1 phase cells in 100 and 200μmol/L FEN groups were significantly elevated compared with that in the control group (allP<0.05). Bioinformatics prediction showed that FEN may play the role against lung adenocarcinoma mainly by activating PPARγ and PPARα receptors, and regulating the expressions of 31 core target molecules such as CCND1 the cell cycle arrest of A549 cells at G1 phase by FEN may be mainly related to down-regulation of CCND1 expression. qRT-PCR and Western blotting verified that FEN could significantly inhibit the gene and protein expression of CCND1 compared with that in the control group (all P<0.05) . Conclusion Fenofibrate may inhibit the proliferation of A549 cells by down-regulating CCND1 expression, and thereby arrest the cell cycle in G1 phase.
万方数据