摘要
目的探讨信号转导和转录激活因子1(STAT1)在非小细胞肺癌中的表达及对IL-2抗肿瘤作用的影响。方法回顾性选取2018年1月至2020年1月丽水市中心医院20例非小细胞肺癌患者外科手术切除的肺癌组织标本及距离肿瘤组织5 cm以上正常肺组织标本,采用Western blot法检测两种标本中STAT1蛋白表达水平。利用慢病毒载体Lenti-增强绿色荧光蛋白(EGFP)或Lenti-STAT1转染非小细胞肺癌SPC-A-1细胞,将细胞分为空白组、Lenti-EGFP对照组和Lenti-STAT1组。采用平板克隆实验观察细胞克隆形成率。不同浓度(0、20、100、500 U/mL)IL-2处理细胞后,采用细胞计数试剂盒法检测细胞增殖活性,Transwell法检测0、100 U/mL IL-2处理后细胞迁移和侵袭能力,流式细胞仪分析0、100 U/mL IL-2处理后细胞凋亡百分比。观察IL-2对3组细胞抗肿瘤作用的影响。采用Western blot法检测3组细胞磷酸化STAT1(p-STAT1)、细胞间黏附分子-1(ICAM-1)及增殖细胞核抗原(PCNA)蛋白表达水平。结果肺癌组织中STAT1蛋白表达水平低于正常组织(P<0.001)。Lenti-STAT1组细胞增殖活性、迁移和侵袭能力均低于Lenti-EGFP对照组,细胞凋亡百分比高于Lenti-EGFP对照组,差异均有统计学意义(均P<0.05)。用20、100、500 U/mL IL-2处理后,Lenti-STAT1组细胞增殖均低于Lenti-EGFP对照组(均P<0.05)。用100 U/mL IL-2处理后,Lenti-STAT1组细胞迁移和侵袭能力均低于Lenti-EGFP对照组,Lenti-STAT1组细胞凋亡百分比高于Lenti-EGFP对照组,差异均有统计学意义(均P<0.05)。此外,IL-2未处理的Lenti-STAT1组细胞p-STAT1蛋白表达水平高于Lenti-EGFP对照组,ICAM-1蛋白表达水平低于Lenti-EGFP对照组,差异均有统计学意义(均P<0.05)。用100 U/mL IL-2处理后,Lenti-STAT1组细胞p-STAT1蛋白表达水平高于Lenti-EGFP对照组,PCNA蛋白表达水平低于Lenti-EGFP对照组,差异均有统计学意义(均P<0.05)。结论STAT1在非小细胞肺癌组织中表达明显下调,可抑制细胞增殖、迁移和侵袭,促进细胞凋亡,增强IL-2在肺癌细胞中的抗肿瘤效应。因此,STAT1与IL-2的联合治疗可能是未来肺癌治疗的一种可行方法。
Objective To explore the expression of signal transducer and activator of transcription 1(STAT1)in nonsmall cell lung cancer(NSCLC)and its influence on the anti-tumor effect of interleukin-2(IL-2).Methods Lung cancer tissue and normal lung tissues which more than 5 cm away from the tumor tissues were collected from 20 NSCLC patients admitted in Lishui Central Hospital From January 2018 to January 2020,and the expression of STAT1 in both samples were detected by Western blot.NSCLC SPC-A-1 cells were transfected with Lenti-STAT1 or Lenti-enhanced green fluorescent protein(EGFP)by lentiviral vectors,and the cells were divided into the blank group,Lenti-EGFP control group and Lenti-STAT1 group.The cell clone formation rate was tested by plate cloning assay.After treatment with 0,20,100 and 500 U/mL IL-2,the cell proliferation was detected by CCK-8 assay.The migration and invasion ability of cells after treatment with 0 and 100 U/mL IL-2 was detected by Transwell assay.The percentage of cell apoptosis after treatment with 0 and 100 U/mL IL-2 was analyzed by flow cytometry.The influence of IL-2 on the anti-tumor effect of 3 groups of cells was observed.The expression levels of phospho STAT1(p-STAT1),intercellular cell adhesion molecule-1(ICAM-1)and proliferating cell nuclear antigen(PCNA)in 3 groups were detected by Western blot.Results The expression level of STAT1 protein in lung cancer tissues was lower than that in normal tissues(P<0.001).The cell proliferation,migration and invasion ability of the Lenti-STAT1 group were lower than those of the Lenti-EGFP control group,and the percentage of cell apoptosis was higher than that of the Lenti-EGFP control group(all P<0.05).After treatment with 20,100 and 500 U/mL IL-2,the cell proliferation of the Lenti-STAT1 group was lower than that of the Lenti-EGFP control group(all P<0.05).After treatment with 100 U/mL IL-2,the migration and invasion ability of Lenti-STAT1 group were lower than those of Lenti-EGFP control group,and the percentage of cell apoptosis of the Lenti-STAT1 group was higher than that of the Lenti-EGFP control group(all P<0.05).Furthermore,the expression level of p-STAT1 protein in the Lenti-STAT1 group without IL-2 treatment was higher than that of the Lenti-EGFP control group,and the expression level of ICAM-1 protein was lower than that of the Lenti-EGFP control group,and the differences were statistically significant(all P<0.05).After treatment with 100 U/mL IL-2,the expression level of p-STAT1 protein in the Lenti-STAT1 group was higher than that of the Lenti-EGFP control group,and the expression level of and PCNA protein was lower than that of the Lenti-EGFP control group(all P<0.05).Conclusion The expression of STAT1 is significantly down-regulated in NSCLC tissues,which can inhibit cell proliferation,migration and invasion,promote cell apoptosis,and enhance the anti-tumor effect of IL-2 in lung cancer cells.Therefore,the combination of STAT1 and IL-2 may be a viable approach for the treatment of lung cancer in the future.
作者
赵嘉璐
孙蕾
蓝秀
熊雪芳
李伟文
ZHAO Jialu;SUN Lei;LAN Xiu;XIONG Xuefang;LI Weiwen(Cancer Center,Lishui Central Hospital,Lishui 323000,China)
出处
《浙江医学》
CAS
2024年第12期1239-1244,1279,I0003,共8页
Zhejiang Medical Journal
基金
丽水市科技计划项目(2020SJZC061)。
