Chlorpromazine cooperatively induces apoptosis with tyrosine kinase inhibitors in EGFR-mutated lung cancer cell lines and restores the sensitivity to gefitinib in T790M-harboring resistant cells
Chlorpromazine cooperatively induces apoptosis with tyrosine kinase inhibitors in EGFR-mutated lung cancer cell lines and restores the sensitivity to gefitinib in T790M-harboring resistant cells
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DOI：
10.1016/j.bbrc.2022.08.010
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发表时间：
2022-08-19
影响因子：
3.1
通讯作者：
Matsumura，Itaru
中科院分区：
文献类型：
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作者：
Fujiwara，Ryosuke;Taniguchi，Yasuhiro;Matsumura，Itaru
我们之前报道过，抗精神病药物氯丙嗪（CPZ）抑制酪氨酸激酶受体（RTK）胞内吞噬和细胞内转运所必需的网格蛋白包被囊泡（ccv）的形成，通过扰乱这些分子的细胞内定位，抑制RTK突变（KIT D816V或FLT3-ITD）的急性髓系白血病细胞的生长/存活。在这里，我们研究了这些发现是否适用于表皮生长因子受体（EGFR）。CPZ剂量依赖性地抑制非小细胞肺癌（NSCLC）细胞系PC9的生长/存活，PC9含有EGFR激活（EGFR外显子19缺失）。此外，CPZ不仅能抑制gefitinib （GEF）耐药PC9ZD细胞的生长/存活，还能恢复其对GEF的敏感性。此外，CPZ克服了HCC827GR细胞中Met扩增引起的GEF抗性。对于CPZ诱导的生长抑制机制，我们发现CPZ虽然对EGFR的磷酸化几乎没有影响，但能有效降低ERK和AKT的磷酸化。当与曲美替尼（MEK抑制剂）、达非尼（RAF抑制剂）和依维莫司（mTOR抑制剂）联合使用时，CPZ与依维莫司协同抑制PC9ZD细胞的生长，而与曲美替尼或达非尼不协同抑制。免疫荧光染色显示EGFR呈核周模式，并与晚期核内体标记物Rab11强烈共定位。但CPZ处理后，EGFR在细胞内分布不均匀，与早期核内体标记物Rab5和EEA1的共定位更加明显，表明CPZ破坏了EGFR的细胞内转运。这些结果表明CPZ具有治疗EGFR突变NSCLC的潜力，其机制不同于传统TKIs单独或与其他药物联合使用。
We previously reported that the antipsychotic drug chlorpromazine (CPZ), which inhibits the formation of clathrin-coated vesicles (CCVs) essential for endocytosis and intracellular transport of receptor tyrosine kinase (RTK), inhibits the growth/survival of acute myeloid leukemia cells with mutated RTK (KIT D816V or FLT3-ITD) by perturbing the intracellular localization of these molecules. Here, we examined whether these findings are applicable to epidermal growth factor receptor (EGFR). CPZ dose-dependently inhibited the growth/survival of the non-small cell lung cancer (NSCLC) cell line, PC9 harboring an EGFR-activating (EGFR exon 19 deletion). In addition, CPZ not only suppressed the growth/survival of gefitinib (GEF)-resistant PC9ZD cells harboring T790 M, but also restored their sensitivities to GEF. Furthermore, CPZ overcame GEF resistance caused by Met amplification in HCC827GR cells. As for the mechanism of CPZ-induced growth suppression, we found that although CPZ hardly influenced the phosphorylation of EGFR, it effectively reduced the phosphorylation of ERK and AKT. When utilized in combination with trametinib (a MEK inhibitor), dabrafenib (an RAF inhibitor), and everolimus (an mTOR inhibitor), CPZ suppressed the growth of PC9ZD cells cooperatively with everolimus but not with trametinib or dabrafenib. Immunofluorescent staining revealed that EGFR shows a perinuclear pattern and was intensely colocalized with the late endosome marker, Rab11. However, after CPZ treatment, EGFR was unevenly distributed in the cells, and colocalization with the early endosome marker Rab5 and EEA1 became more apparent, indicating that CPZ disrupted the intracellular transport of EGFR. These results suggest that CPZ has therapeutic potential for NSCLC with mutated EGFR by a novel mechanism different from conventional TKIs alone or in combination with other agents.