【摘要】 目的 分析髓细胞白血病1（MCL-1）的表达与非小细胞肺癌（NSCLC）患者生存的关系，探讨过表达MCL-1对NSCLC细胞生物学行为的影响和在NSCLC进展中的作用。方法 分别利用GEPIA、Kaplan-Meier Plotter和HPA数据库分析MCL-1在肺癌、肺腺癌（LUAD）和肺鳞状细胞癌（LUSC）各分期的表达及其与患者生存预后的关系。通过STRING数据库绘制与MCL-1相互关联蛋白网络图，并进行GO和KEGG富集分析。采用慢病毒转染，构建MCL-1过表达HCC827^MCL-1以及空载对照HCC827^NC的NSCLC模型。平板克隆形成实验分析细胞的增殖及克隆形成能力；Transwell和划痕实验检测细胞侵袭、迁移能力；MTT法测定细胞生长情况、倍增时间及对顺铂（DDP）和多西他赛（TXT）的敏感性。结果 MCL-1在LUAD和LUSC各分期中均呈高表达，且随着肿瘤的恶化，MCL-1表达有升高的趋势。Kaplan-Meier Plotter数据库分析结果显示，MCL-1高表达患者中位总生存期分别为64.1、62.0和64.1个月，MCL-1低表达患者中位生存期分别为76.0、79.3和75.4个月，P值分别为0.003、<0.001和0.041。HPA数据库分析结果显示，肺癌、LUAD和LUSC MCL-1高表达患者的5年生存率分别为38%、30%和41%,低表达患者分别为51%、42%和51%,P值分别为0.024、0.029和0.031。GO和KEGG分析结果显示，MCL-1主要与细胞凋亡和耐药性等生物学过程有关。成功构建了过表达MCL-1的HCC827细胞模型。克隆形成实验结果显示，在培养10 d后，HCC827^NC和HCC827^MCL-1组克隆细胞数分别为（274±10）和（315±9）个，t=5.203,P=0.007。HCC827^NC细胞和HCC827^MCL-1细胞的倍增时间分别为（26.38±2.17）和（17.15±1.63） h,t=5.887,P=0.004。Transwell实验结果显示，48 h后HCC827^NC和HCC827^MCL-1组穿过小室的细胞数分别为（225±6）和（389±34）个，t=7.269,P=0.002。划痕实验结果显示，HCC827^NC组和HCC827^MCL-1组48 h划痕愈合率分别为（20.46±0.10）%和（30.55±0.29）%,t=56.820,P<0.001。MTT实验结果显示，DDP作用于HCC827^NC细胞72 h的抑制率为（69.74±0.80）%,HCC827^MCL-1细胞为（58.59±3.16）%,t=5.940,P=0.004;TXT作用于HCC827^NC细胞72 h的抑制率为（54.15±0.42）%,HCC827^MCL-1细胞为（43.88±0.55）%,t=25.870,P<0.001。结论 MCL-1表达水平上调和NSCLC预后不良有关，MCL-1高表达可促进NSCLC的恶性进程，降低其对化疗药物的敏感性，MCL-1或可作为NSCLC新的潜在预后生物标志物。更多还原

【Abstract】 Objective To analyze the relationship between the expression of myeloid cell leukemia-1（MCL-1） and the survival of patients with non-small cell lung cancer（NSCLC） and the effect of overexpression of MCL-1 on the biological behavior of NSCLC cells,and explore the role of MCL-1 in tumor progression.Methods GEPIA,Kaplan-Meier Plotter and HPA databases were used to analyze the expression of MCL-1 in lung cancer,lung adenocarcinoma（LUAD） and lung squamous cell carcinoma（LUSC） stage and its relationship with patient survival prognosis.The protein-protein interaction analysis,GO and KEGG enrichment analysis were performed on STRING database.The lentiviral transfection was used to construct of NSCLC models with overexpressing of MCL-1 in HCC827^MCL-1 and empty control HCC827NC.The proliferation and clonogenic ability of the cells were studied by plate clone formation assay.Transwell and scratch test were performed to measure the invasion and migration of cells.The growth,doubling time and sensitivity of cells to cisplatin（DDP） and docetaxel（TXT） were determined by MTT method.Results MCL-1 was highly expressed in all stages of LUAD and LUSC,and its expression tended to increase as the tumor deteriorated.The Kaplan Meier Plotter database analysis results showed that the median survival time of patients with high MCL-1 expression was 64.1,62.0 and 64.1months,respectively,while the median survival time of patients with low MCL-1 expression was 76.0,79.3 and 75.4months,respectively,with P values of 0.003,<0.001 and 0.041.The analysis results of the HPA database showed that the 5-year survival rates of lung cancer,LUAD and LUSC patients with high expression of MCL-1 were 38%,30% and41%,respectively,while those with low expression were 51%,42% and 51%,respectively,with P values of 0.024,0.029and 0.031,respectively.GO and KEGG analysis results showed that MCL-1 was mainly related to biological processes such as cell apoptosis and drug resistance.We successfully constructed a HCC827 cell model overexpressing MCL-1.The results of the clone formation experiment showed that after 10 days of cultivation,the number of cloned cells in the HCC827^NC and HCC827^MCL-1 groups was 274±10 and 315±9,respectively,t=5.203,P=0.007.The doubling times of HCC827^NC cells and HCC827^MCL-1 cells were（26.38 ± 2.17） and（17.15±1.63） hours,respectively,t=5.887,P=0.004.The Transwell experiment results showed that after 48 hours,the number of cells passing through the compartments in the HCC827^NC and HCC827^MCL-1 groups was 225±6 and 389±34,respectively,t=7.269,P=0.002.The scratch experiment results showed that the 48 hour scratch healing rates of the HCC827^NC group and HCC827^MCL-1 group were（20.46±0.10） % and（30.55±0.29） %,respectively,t=56.820,P<0.001.The MTT experiment results showed that the inhibitory rate of DDP on HCC827^NC cells after 72 hours was（69.74±0.80）%,while that of HCC827^MCL-1 cells was（58.59 ±3.16） %,t=5.940,P=0.004.The inhibitory rate of TXT on HCC827^NC cells for 72 hours was（54.15 ±0.42） %,while that on HCC827^MCL-1 cells was（43.88±0.55） %,t=25.870,P<0.001.Conclusions The up-regulation of MCL-1 expression level is associated with poor prognosis of NSCLC.The high expression of MCL-1 promotes the malignant process in NSCLC and can reduce the sensitivity of cells to drugs.MCL-1 may be used as a new prognostic potential biomarker for NSCLC.更多还原