【摘 要】
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目的:通过RNA测序分析比较亲本PC-9细胞和厄洛替尼获得性耐药PC-9细胞(PC-9/ER)表达谱的差异,揭示非小细胞肺癌(NSCLC)表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)耐药的潜在机制.方法:采用间歇诱导方法,建立PC-9/ER耐药细胞株,通过MTF实验绘制厄洛替尼药物浓度-细胞存活率曲线.通过RNA测序分析PC-9/ER细胞的差异表达基因并进行GO和KEGG功能富集分析;通过qRT-PCR进一步筛选可能参与EGFR-TKI耐药的潜在基因或可能的靶点.结果:厄洛替尼对PC-9/ER细胞
【机 构】
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西安交通大学第一附属医院人才高地科研平台,陕西西安710061;西安交通大学第一附属医院人才高地科研平台,陕西西安710061;西安交通大学第一附属医院普通外科,陕西西安710061;西安交通大学第一
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目的:通过RNA测序分析比较亲本PC-9细胞和厄洛替尼获得性耐药PC-9细胞(PC-9/ER)表达谱的差异,揭示非小细胞肺癌(NSCLC)表皮生长因子受体酪氨酸激酶抑制剂(EGFR-TKI)耐药的潜在机制.方法:采用间歇诱导方法,建立PC-9/ER耐药细胞株,通过MTF实验绘制厄洛替尼药物浓度-细胞存活率曲线.通过RNA测序分析PC-9/ER细胞的差异表达基因并进行GO和KEGG功能富集分析;通过qRT-PCR进一步筛选可能参与EGFR-TKI耐药的潜在基因或可能的靶点.结果:厄洛替尼对PC-9/ER细胞增殖的抑制率显著低于PC-9细胞的抑制率,PC-9/ER细胞的耐药指数为41.92.与PC-9细胞相比,RNA测序PC-9/ER细胞筛选出1028个差异表达基因,其中720个基因表达上调,308个基因表达下调,而且差异表达基因显著富集在PI3K-AKT通路和癌症通路.qRT-PCR验证差异表达基因的转录水平与测序结果基本一致.结论:ST6GALNAC3、CYP1A1、PAPPA2、INHBE和ACSS3等基因可能参与EGFR-TKI的耐药过程,针对PC-9/ER细胞差异表达基因的后续实验可能为将来改善NSCLC的靶向治疗进一步提供实验和理论依据.
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