着丝粒蛋白F在非小细胞肺癌组织中的表达及其对A549细胞生物学行为的影响研究

背景非小细胞肺癌(NSCLC)占肺癌的85%左右,由于易错失最佳手术切除时机,且放化疗效果不佳,导致生存率偏低。而着丝粒蛋白F(CENPF)是在多种癌症中高表达的分子,探讨其对肺癌细胞生物学行为的影响,将有助于NSCLC的靶向治疗。目的探讨CENPF在NSCLC组织中的表达及其对A549细胞生物学行为的影响。方法收集2018年10月—2019年10月于西安医学院第一附属医院行外科切除手术的26例NSCLC患者的癌组织、癌旁组织标本。采用Western blotting法检测NSCLC组织、癌旁组织中CENPF表达水平。取生长良好的对数期A549细胞,将其分为对照组、阴性对照组、CENPF抑制组,其中对照组A549细胞不进行转染处理,阴性对照组、CENPF抑制组A549细胞使用Lipofectamine 2000转染试剂分别转染阴性对照(NC)-干扰性小核糖核酸(si RNA)、CENPF-si RNA。采用实时荧光定量聚合酶链式反应(q PCR)法检测三组A549细胞中CENPF m RNA表达水平,细胞计数试剂盒8(CCK-8)法检测A549细胞增殖情况吸光度(OD)值],流式细胞仪检测A549细胞凋亡情况、A549细胞周期,Western blotting法检测A549细胞中CENPF、细胞周期蛋白D1(cyclin D1)、B淋巴细胞瘤-2基因相关X蛋白(Bax)表达水平。结果 NSCLC组织中CENPF表达水平高于癌旁组织(P <0.05)。CENPF抑制组A549细胞中CENPF m RNA表达水平低于对照组、阴性对照组(P<0.05)。CENPF抑制组A549细胞OD值低于对照组、阴性对照组(P<0.05)。CENPF抑制组A549细胞凋亡率高于对照组、阴性对照组(P <0.05)。CENPF抑制组G0/G1期A549细胞所占比例高于对照组、阴性对照组,S期、G2/M期A549细胞所占比例低于对照组、阴性对照组(P <0.05)。CENPF抑制组A549细胞中CENPF、cyclin D1表达水平低于对照组、阴性对照组,Bax表达水平高于对照组、阴性对照组(P <0.05)。结论 CENPF在NSCLC组织中表达升高,其可通过影响cyclin D1、Bax表达水平来影响A549细胞周期进展及增殖、凋亡过程;下调其表达可将A549细胞周期阻滞于G0/G1期,抑制细胞增殖并诱导其凋亡。

Expression of Centromere Protein F in Non-small Cell Lung Cancer Tissues and Its Effect on the Biological Behavior of A549 Cells

Background Non-small cell lung cancer(NSCLC)accounts for about 85%of lung cancer,with low survival rate,because it is easy to miss the best resection period and the effect of radiotherapy and chemotherapy is poor.Centromere protein F(CENPF)is a highly expressed molecule in multiple cancers,and exploring its effect on the biological behavior of lung cancer cells will contribute to NSCLC targeted therapy.Objective To investigate the expression of CENPF in NSCLC tissues and its effect on the biological behavior of A549 cells.Methods The cancer tissue and precancerous lesions of 26 NSCLC patients who underwent surgical resection in the First Affiliated Hospital of Xi’an Medical University from October 2018 to October 2019 were collected.Western blotting was used to detect the expression level of CENPF in NSCLC tissues and precancerous lesions.The well-growing A549 cells in log phase were divided into control group,negative control group and CENPF inhibition group.A549 cells in the control group were not transfected,and A549 cells in the negative control group and CENPF inhibition group were transfected with NC-siRNA and CENPF-siRNA respectively with Lipofectamine 2000 transfection reagent.The expression of CENPF mRNA in A549 cells was detected by real-time fluorescence quantitative PCR(qPCR),the proliferation of A549 cellsabsorbance(OD)value]was detected by cell count kit 8(CCK-8)method,the apoptosis and cell cycle of A549 cells were detected by flow cytometry,the expression levels of CENPF,cyclinD1 and B-cell lymphoma-2 associated X protein(Bax)in A549 cells were detected by Western blotting.Results The expression level of CENPF in the NSCLC tissue was significantly higher than those in the precancerous lesions(P<0.05).Compared with the control group and the negative control group,the expression level of CENPF mRNA in A549 cells of the CENPF inhibition group was significantly lower(P<0.05).The OD value of A549 cells in the CENPF inhibition group was significantly lower than that in the control group and the negative control group(P<0.05).The apoptosis rate of A549 cells in the CENPF inhibition group was higher than that in the control group and the negative control group(P<0.05).The proportion of A549 cells in the G0/G1 phase of the CENPF inhibition group was higher than that of the control group and the negative control group,and the proportion of A549 cells in the S phase and G2/M phase was lower than that of the control group and the negative control group(P<0.05).The expression levels of CENPF and cyclinD1 in A549 cells of the CENPF inhibition group were lower than those of the control group and the negative control group,and the expression levels of Bax were higher than those of the control group and the negative control group(P<0.05).Conclusion The expression of CENPF is increased in NSCLC tissues,and it can affect A549 cell cycle progression,proliferation,and apoptosis by affecting the expression levels of cyclinD1 and Bax.Down-regulating the expression of CENPF can block the A549 cell cycle in the G0/G1 phase,inhibit cell proliferation and induce its apoptosis.