吉非替尼治疗晚期非小细胞肺癌耐药前后患者血清miRNAs的表达差异  

Differential expression of serum miRNAs in patients with advanced non-small cell lung cancer treated by gifitinib before and after acquiring drug resistance

作　　者：黄谦[1] 李代强[2] HUANG Qian;LI Daiqiang(Department of Basic Medicine,Yueyang Vocational Technical College,Yueyang Hunan 414000;Department of Pathology,Second Xiangya Hospital,Central South Univesity,Changsha 410011,China)

机构地区：[1]岳阳职业技术学院基础医学部,湖南岳阳414000 [2]中南大学湘雅二医院病理科,长沙410011

出　　处：《中南大学学报（医学版）》2018年第12期1288-1293,共6页Journal of Central South University ：Medical Science

摘　　要：目的:研究吉非替尼治疗晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)发生继发性耐药前后患者血清microRNA(miRNAs)表达的差异。方法:选择2013年6月至2015年6月岳阳市职业技术学院附属医院收治的4例经吉非替尼常规治疗出现获得性耐药的晚期NSCLC患者作为研究对象。收集研究对象治疗前和出现获得性耐药后的血清样本。采用miRNA芯片技术对血清样本中miRNAs表达谱进行检测,实时RT-PCR对差异性表达显著的miRNAs进行定量验证。针对筛选出的miRNAs构建抑制剂和拟似剂并转染至肺癌细胞中,检测肺癌细胞对敏感性的变化。结果:发生继发性耐药前后患者血清miRNAs表达发生明显改变。24个miRNAs在治疗前和出现获得性耐药后的NSCLC患者血清中表达具有明显的差异(表达差异≥2倍);其中19个miRNAs在出现获得性耐药后明显上调,5个miRNAs在出现获得性耐药后明显下调,与治疗前相比差异均有统计学意义(均P<0.05)。MiR-21在出现获得性耐药后NSCLC患者的血清中的表达水平为治疗前的10倍多。RT-PCR检测miR-21的结果与miRNA芯片结果相符合。增强肺癌细胞miR-21的表达能够提高肺癌细胞对吉非替尼的半数抑制浓度(half maximal inhibitory concentration,IC50),而抑制肺癌细胞miR-21的表达能够降低肺癌细胞对吉非替尼的IC50(均P<0.05)。结论:NSCLC经吉非替尼治疗发生耐药前后患者血清中存在差异表达的miRNAs,其中高表达的miR-21可能与吉非替尼的获得性耐药有关。Objective: To explore the diff erential expression of serum miRNAs in patients of advanced nonsmall cell lung cancer (NSCLC) treated by gifi tinib before and aft er acquiring drug resistance. Methods: A total of 4 patients with advanced NSCLC from Affiliated Hospital of Yueyang Vocational Technical College, who acquired drug resistance during gefitinib therapy from June 2013 to June 2015, were enrolled. Serum samples were collected before treatment and after acquiring drug resistance. MicroRNA (miRNA) microarray was used to assess the levels and compositions of miRNAs in serum. Real-time RT-PCR was used to validate the results of miRNAs with significant differences in expression. The candidate miRNAs inhibitors and mimics were transfected into lung cancer cells by liposome, and the sensitivity of lung cancer cells to gifitinib was detected. Results: The miRNA microarray showed that there were significantly differential expression of miRNAs in serum of NSCLC patients after acquiring drug resistance, and 24 miRNAs were changed in more than 2-fold. Among them, 19 miRNAs were up-regulated and 5 miRNAs were down- regulated (both P<0.05). Especially, the expression of miR-21 in serum of NSCLC patients after obtaining resistance was up-regulated more than 10-fold compared with that before treatment. The results of RT-PCR was consistent with the results of miRNA microarray. The up-regulation of miR-21 in lung cancer cells could elevate the half maximal inhibition concentration (IC50) of gefitinib, and the down-regulation of miR-21 in lung cancer cells could reduce the IC50 of gefitinib (both P<0.05). Conclusion: There is differential expression of miRNAs in serum of NSCLC patients before treatment and after acquiring drug resistance during gefitinib therapy. The up-regulation of miR-21 may be involved in regulating the acquiring drug resistance of gefitinib.

关 键 词：非小细胞肺癌 吉非替尼 获得性耐药 MIRNA芯片 

分 类 号：R734.2[医药卫生—肿瘤]