信号转导与转录激活因子3下调DNAJB6在肺癌生物学行为中的机制  

Role of downregulation of DNAJB6 by signal transducer and activator of transcription 3 in biological behaviors of non-small cell lung cancer

作　　者：朱定强 刘相燕[2] 陈祥玉 史墨 任志超 马官强 于洋 Zhu Dingqiang;Liu Xiangyan;Chen Xiangyu;Shi Mo;Ren Zhichao;Ma Guanqiang;Yu Yang(Cheeloo College of Medicine,Shandong University,Jinan 250000,China;Department of Thoracic Surgery,Shandong Provincial Hospital Affiliated to Shandong First Medical University,Jinan 250000,China;Department of Thoracic Surgery,Shandong Provincial Hospital,Jinan 250000,China)

机构地区：[1]山东大学齐鲁医学院,济南250000 [2]山东省立医院胸外科,济南250000 [3]山东第一医科大学附属省立医院胸外科,济南250000

出　　处：《中华实验外科杂志》2023年第4期675-677,共3页Chinese Journal of Experimental Surgery

基　　金：山东省医药卫生科技发展计划(202004021371)。

摘　　要：目的探讨非小细胞肺癌(NSCLC)细胞中信号转导与转录激活因子3(STAT3)与DNA结合蛋白B6(DNAJB6)的关系及铁死亡对NSCLC生物学行为的影响。方法采用细胞计数试剂盒(CCK-8)细胞增殖试验和细胞划痕实验对非小细胞肺癌细胞系A549和H1650进行细胞增殖能力检测。通过慢病毒转染A549和H1650细胞系,检测NSCLC细胞的迁移和侵袭能力。蛋白质印迹法(Western blot)检测NSCLC细胞系A549和H1650各项靶蛋白的表达,组间比较采用t检验,多组比较采用方差分析。结果Western blot结果显示在膜上约80 kDa处,肿瘤组织蛋白条带较正常相邻组织颜色更深(P<0.01)。反转录-聚合酶链反应(RT-PCR)分析结果显示,肿瘤组织中STAT3/DNAJB6 mRNA与GAPDH mRNA的比值高于正常相邻组织(1.184±0.168比0.378±0.136)差异有统计学意义(P<0.01)。CCK-8实验结果,在24、48、72、96 h时,STAT3组的NSCLC细胞系A549增殖率比相应对照组高[(1.000±0.011)%、(2.338±0.021)%、(3.324±0.010)%、(4.270±0.008)%比(1.00±0.012)%、(1.789±0.010)%、(2.891±0.060)%、(3.761±0.017)%],差异有统计学意义(P<0.01);NSCLC细胞系H1650增殖率比相应对照组高[(1.000±0.008)%、(1.706±0.020)%、(2.956±0.096)%、(3.755±0.015)%比(1.000±0.004)%、(1.484±0.004)%、(2.336±0.008)%、(3.074±0.107)%],差异有统计学意义(P<0.01)。细胞划痕实验结果:过表达STAT3后,NSCLC细胞系A549和H1650增殖能力均显著增强(P<0.01)。病毒转染A549和H1650细胞系结果:过表达STAT3后NSCLC细胞的迁移和侵袭能力增强(P<0.01)。Western blot结果:DNAJB6表达减少,铁死亡相关蛋白xCT和GPX4表达增加(表达水平与铁死亡呈负相关,P<0.01)。结论过表达STAT3可抑制癌细胞铁死亡。Objective To explore the relationship between signal transducer and activator of transcription 3(STAT3)and DNAJB6 in non-small cell lung cancer(NSCLC)cell lines,and the effect of ferroptosis on NSCLC biological behaviors.Methods Cell counting kit-8(CCK-8)cell proliferation assay and cell scratch assay were performed on NSCLC cell lines A549 and H1650 to test the proliferation ability of NSCLC cells.The migration and invasion ability of NSCLC cells was tested through lentivirus transfection of A549 and H1650 cell lines.Western blotting analysis was performed on NSCLC cell lines A549 and H1650.The t-test was used for comparison between two groups in data analysis,while analysis of variance(ANOVA)was used for comparisons among more than two groups.Results The Western blotting results indicated that at approximately 80 kDa on the membrane,the protein band in tumor tissue was more intense in color than normal adjacent tissue(P<0.01).The reverse transcription-polymerase chain reaction(RT-PCR)analysis showed that the ratio of STAT3/DNAJB6 mRNA to GAPDH mRNA in tumor tissue was higher than that in adjacent normal tissue(1.184±0.168 vs.0.378±0.136),with statistical significance(P<0.01).The CCK-8 experiment results showed that the proliferation rate of NSCLC cell lines A549 and H1650 in the STAT3 group was higher than that in the corresponding control group at 24,48,72,and 96 h.For A549 cells,the proliferation rate was(1.000±0.011)%,(2.338±0.021)%,(3.324±0.010)%,and(4.270±0.008)%,vs.(1.000±0.012)%,(1.789±0.010)%,(2.891±0.060)%,and(3.761±0.017)%,respectively,with statistical significance(P<0.01).For H1650 cells,the proliferation rate was(1.000±0.008)%,(1.706±0.020)%,(2.956±0.096)%,and(3.755±0.015)%,vs.(1.000±0.004)%,(1.484±0.004)%,(2.336±0.008)%,and(3.074±0.107)%,respectively,with statistical significance(P<0.01).The cell scratch assay results showed that after overexpression of STAT3,the proliferation ability of NSCLC cell lines A549 and H1650 was significantly enhanced(P<0.01).Viral transfection of A549

关 键 词：非小细胞肺癌 信号转导与转录激活子3 DNA结合蛋白B6 铁死亡 生物学行为 

分 类 号：R734.2[医药卫生—肿瘤]