吉非替尼获得性耐药非小细胞肺癌细胞中差异表达环状RNA分析
发布时间:2019-05-30 07:21
【摘要】:目的 :建立吉非替尼获得性耐药非小细胞肺癌细胞株,并筛选和分析耐药前后差异表达的环状RNA(circular RNA,circ RNA)。方法:利用吉非替尼敏感性非小细胞肺癌细胞株H292(简称为H292_S),采用逐步加药法建立吉非替尼获得性耐药株(命名为H292_R)。CCK-8法检测吉非替尼对2种细胞增殖的抑制作用;RNA测序(RNA sequencing,RNA-seq)法筛选2种细胞中差异表达的circRNA;实时荧光定量PCR法验证RNA-seq结果,并结合统计学和生物信息学方法对差异circRNA来源基因的生物学功能进行分析。结果:吉非替尼对H292_S和H292_R细胞增殖的半数抑制浓度分别为(108.63±0.32)nmol/L和(982.37±2.62)nmol/L,2者间差异有统计学意义(P值均0.05),耐药细胞H292_R对吉非替尼的耐药指数为9.04。2种细胞中存在36个差异表达circRNA,其中耐药细胞中24个circRNA表达上调(P值均0.05),12个circRNA表达下调(P值均0.05);GO富集分析发现,异常表达的circRNA主要涉及调节细胞生长增殖、蛋白转运和基因转录等生物学过程;KEGG通路分析发现,异常circRNA的信号通路主要涉及细胞质DNA感受通路、核因子κB(nuclear factor-kappa B,NF-κB)、胞吞和凋亡等通路。实时荧光定量PCR法验证2种细胞中差异表达circRNA hsa_circ_0000567和hsa_circ_0000620的表达水平差异,结果显示耐药细胞中这2个circRNA水平均明显上调(P值均0.05),与RNA-seq结果相符。结论:筛选出吉非替尼获得性耐药相关的差异表达circRNA,这有助于更深入地阐明非小细胞肺癌耐药的机制,并可能为其早期诊断提供生物学标志。
[Abstract]:Objective: to establish a gefitinib acquired drug-resistant non-small cell lung cancer cell line and screen and analyze the differentially expressed cyclic RNA (circular RNA,circ RNA). Before and after drug resistance. Methods: Gifetinib sensitive non-small cell lung cancer cell line H292 (abbreviated as H292 鈮,
本文编号:2488632
[Abstract]:Objective: to establish a gefitinib acquired drug-resistant non-small cell lung cancer cell line and screen and analyze the differentially expressed cyclic RNA (circular RNA,circ RNA). Before and after drug resistance. Methods: Gifetinib sensitive non-small cell lung cancer cell line H292 (abbreviated as H292 鈮,
本文编号:2488632
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