安罗替尼可以通过非小细胞肺癌中没有 EGFR T790M 突变的 FGFR1 信号转导克服获得性对 EGFR-TKIs 的耐药性。

Anlotinib can overcome acquired resistance to EGFR-TKIs via FGFR1 signaling in non-small cell lung cancer without harboring EGFR T790M mutation.

机构信息

Department of Respiratory Medicine, The First Affiliated Hospital of Soochow University, Suzhou, China.

Institute of Respiratory Diseases, Soochow University, Suzhou, China.

出版信息

Thorac Cancer. 2020 Jul;11(7):1934-1943. doi: 10.1111/1759-7714.13485. Epub 2020 May 20.

BACKGROUND

Although many studies have defined mechanisms of resistance to EGFR-TKIs, acquired resistance remains the major limitation of monotherapy with EGFR-TKIs.

METHODS

Cell viability was analyzed using a Cell Counting Kit-8 (CCK-8) assay. EGFR T790M mutation was sequenced on a HiSeq 4000 platform. mRNAs from HCC827 and HCC827 gefitinib-resistant (GR) cells were analyzed by genome analyzer-based deep sequencing. The effect of anlotinib on apoptosis and cell cycle arrest of HCC827 GR was detected by fluorescence-activated cell sorting (FACS) analysis. A mouse xenograft model was used to assess the effect of anlotinib on HCC827 GR cells.

RESULTS

The T790M mutation was found in the PC-9 GR cell line but not in the HCC827 GR cell line. Anlotinib could suppress the growth of HCC827 GR cells by inhibiting FGFR1 in vitro and in a mouse xenograft model. Moreover, FGFR1 was overexpressed in HCC827 GR cells, and the knockdown of FGFR1 reversed gefitinib resistance in HCC827 GR cells. Furthermore, anlotinib induced apoptosis and cell cycle arrest in HCC827 GR cells by increasing the activity of Caspase-3.

CONCLUSIONS

FGFR1 overexpression could be the mechanism of EGFR-TKI acquired resistance and anlotinib can suppresse the growth of EGFR-TKI-resistant NSCLC cells without T790M mutation.

背景

尽管许多研究已经确定了对 EGFR-TKIs 产生耐药性的机制，但获得性耐药仍然是 EGFR-TKIs 单药治疗的主要限制。

方法

使用细胞计数试剂盒-8（CCK-8）测定法分析细胞活力。在 HiSeq 4000 平台上对 EGFR T790M 突变进行测序。通过基因组分析仪深度测序分析 HCC827 和 HCC827 吉非替尼耐药（GR）细胞的 mRNA。通过荧光激活细胞分选（FACS）分析检测安罗替尼对 HCC827 GR 细胞凋亡和细胞周期阻滞的影响。使用小鼠异种移植模型评估安罗替尼对 HCC827 GR 细胞的作用。

结果

在 PC-9 GR 细胞系中发现了 T790M 突变，但在 HCC827 GR 细胞系中未发现。安罗替尼可通过体外抑制 FGFR1 和小鼠异种移植模型抑制 HCC827 GR 细胞的生长。此外，FGFR1 在 HCC827 GR 细胞中过表达，并且 FGFR1 的敲低逆转了 HCC827 GR 细胞对吉非替尼的耐药性。此外，安罗替尼通过增加 Caspase-3 的活性诱导 HCC827 GR 细胞凋亡和细胞周期阻滞。