Targeting mTOR to overcome epidermal growth factor receptor tyrosine kinase inhibitor resistance in non-small cell lung cancer cells.
Targeting mTOR to overcome epidermal growth factor receptor tyrosine kinase inhibitor resistance in non-small cell lung cancer cells.
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靶向 mTOR 克服非小细胞肺癌细胞中表皮生长因子受体酪氨酸激酶抑制剂耐药性。
DOI：
10.1371/journal.pone.0069104
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Wu YL
中科院分区：
文献类型：
--
作者：
Fei SJ;Zhang XC;Dong S;Cheng H;Zhang YF;Huang L;Zhou HY;Xie Z;Chen ZH;Wu YL
表皮生长因子受体（EGFR）酪氨酸激酶抑制剂（TKI）已在晚期非小细胞肺癌（NSCLC）中显示出显著的临床获益;然而，耐药仍然是临床实践中的一个严重问题。本研究分析了EGFR TKI敏感和耐药NSCLC细胞系之间的mTOR相关信号通路差异，并研究了在EGFR TKI耐药NSCLC细胞中使用特异性mTOR抑制剂靶向mTOR的可行性。我们选择了四种不同类型的EGFR TKI敏感和耐药的NSCLC细胞：PC 9、PC 9 GR、H1650和H1975作为模型，通过western blot和免疫沉淀检测mTOR相关信号通路的差异，并通过MTT法和流式细胞术评价ku-0063794的抗增殖作用和细胞周期阻滞。在本研究中，我们观察到mTORC 2相关的Akt ser 473-FOXO 1信号通路在基础状态下在耐药细胞中高度活化。体外mTORC 1和mTORC 2激酶活性测定显示，EGFR TKI耐药NSCLC细胞系具有较高的mTORC 2激酶活性，而敏感细胞在基础状态下具有较高的mTORC 1激酶活性。ATP竞争性mTOR抑制剂ku-0063794在敏感和耐药细胞中均显示出显著的抗增殖作用和G1细胞周期阻滞。Ku-0063794在IC 50浓度下有效抑制mTOR和p70 S6 K磷酸化水平;后者是mTORC 1底物，不会上调Akt ser 473磷酸化，这将由雷帕霉素诱导，并导致FOXO 1磷酸化的部分抑制。我们还观察到EGFR TKI敏感和耐药的临床NSCLC肿瘤标本具有更高的总p70 S6 K表达水平和磷酸化p70 S6 K表达水平。我们的研究结果表明，mTORC 2相关信号通路在EGFR TKI耐药细胞中过度活化，使用特异性mTOR抑制剂靶向mTOR可能是EGFR突变型NSCLC患者发生EGFR TKI耐药的良好策略; mTORC 2在EGFR TKI耐药NSCLC细胞中的潜在特异性作用尚不清楚，应进一步研究。
Epidermal growth factor receptor (EGFR) tyrosine kinase inhibitors (TKIs) have shown dramatic clinical benefits in advanced non-small cell lung cancer (NSCLC); however, resistance remains a serious problem in clinical practice. The present study analyzed mTOR-associated signaling-pathway differences between the EGFR TKI-sensitive and -resistant NSCLC cell lines and investigated the feasibility of targeting mTOR with specific mTOR inhibitor in EGFR TKI resistant NSCLC cells. We selected four different types of EGFR TKI-sensitive and -resistant NSCLC cells: PC9, PC9GR, H1650 and H1975 cells as models to detect mTOR-associated signaling-pathway differences by western blot and Immunoprecipitation and evaluated the antiproliferative effect and cell cycle arrest of ku-0063794 by MTT method and flow cytometry. In the present study, we observed that mTORC2-associated Akt ser473-FOXO1 signaling pathway in a basal state was highly activated in resistant cells. In vitro mTORC1 and mTORC2 kinase activities assays showed that EGFR TKI-resistant NSCLC cell lines had higher mTORC2 kinase activity, whereas sensitive cells had higher mTORC1 kinase activity in the basal state. The ATP-competitive mTOR inhibitor ku-0063794 showed dramatic antiproliferative effects and G1-cell cycle arrest in both sensitive and resistant cells. Ku-0063794 at the IC50 concentration effectively inhibited both mTOR and p70S6K phosphorylation levels; the latter is an mTORC1 substrate and did not upregulate Akt ser473 phosphorylation which would be induced by rapamycin and resulted in partial inhibition of FOXO1 phosphorylation. We also observed that EGFR TKI-sensitive and -resistant clinical NSCLC tumor specimens had higher total and phosphorylated p70S6K expression levels. Our results indicate mTORC2-associated signaling-pathway was hyperactivated in EGFR TKI-resistant cells and targeting mTOR with specific mTOR inhibitors is likely a good strategy for patients with EGFR mutant NSCLC who develop EGFR TKI resistance; the potential specific roles of mTORC2 in EGFR TKI-resistant NSCLC cells were still unknown and should be further investigated.
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DOI：
10.1158/1078-0432.ccr-09-1837
发表时间：
2010-02-01
期刊：
Clinical cancer research : an official journal of the American Association for Cancer Research
影响因子：
--
作者：
Ghobrial IM;Roccaro A;Hong F;Weller E;Rubin N;Leduc R;Rourke M;Chuma S;Sacco A;Jia X;Azab F;Azab AK;Rodig S;Warren D;Harris B;Varticovski L;Sportelli P;Leleu X;Anderson KC;Richardson PG
通讯作者：
Richardson PG
影响因子：
45.3
作者：
Baselga, J;Rischin, D;Albanell, J
通讯作者：
Albanell, J
影响因子：
7.3
作者：
Guertin, David A.;Sabatini, David M.
通讯作者：
Sabatini, David M.
影响因子：
5.7
作者：
Breuleux M;Klopfenstein M;Stephan C;Doughty CA;Barys L;Maira SM;Kwiatkowski D;Lane HA
通讯作者：
Lane HA
影响因子：
158.5
作者：
Mok, Tony S.;Wu, Yi-Long;Fukuoka, Masahiro
通讯作者：
Fukuoka, Masahiro