Overcoming erlotinib resistance in EGFR mutation-positive lung adenocarcinomas through repression of phosphoglycerate dehydrogenase.
Overcoming erlotinib resistance in EGFR mutation-positive lung adenocarcinomas through repression of phosphoglycerate dehydrogenase.
复制标题
通过抑制磷酸甘油酸脱氢酶克服 EGFR 突变阳性肺腺癌的厄洛替尼耐药性
作者：
Dong JK;Lei HM;Liang Q;Tang YB;Zhou Y;Wang Y;Zhang S;Li WB;Tong Y;Zhuang G;Zhang L;Chen HZ;Zhu L;Shen Y
如何提高表皮生长因子受体酪氨酸激酶抑制剂（EGFR-TKI）的疗效，逆转其耐药性，是EGFR激活突变肺腺癌靶向治疗的主要挑战。磷酸甘油酸脱氢酶（PHGDH）是丝氨酸生物合成的关键酶，在包括肺癌在内的多种癌症中过度表达。PHGDH表达升高与临床肺腺癌患者总体生存率降低相关。在此，我们研究了PHGDH在肺腺癌中对厄洛替尼耐药的作用。方法：采用RNA-Seq技术筛选肺腺癌细胞获得性厄洛替尼耐药所必需的基因。然后通过免疫印迹和qRT-PCR证实厄洛替尼抗性细胞中PHGDH的蛋白和mRNA水平。分别使用细胞培养和肿瘤异种移植小鼠模型检查PHGDH抑制或过表达对厄洛替尼抗性的影响。通过DCFH-DA染色和免疫荧光法检测PHGDH抑制后细胞内ROS水平和DNA损伤标志物γ H2 AX的变化，探讨PHGDH对细胞凋亡的影响。结果如下：我们发现，PHGDH水平显着增加，在肺腺癌PC 9 ER 4和HCC 827 ER 9细胞获得耐药性厄洛替尼。通过siPHGDH转染或NCT-503（一种小分子PHGDH抑制剂）干扰PHGDH协同增强了杀肿瘤作用，并恢复了细胞系和异种移植物中对厄洛替尼的敏感性。PHGDH的过表达导致异种移植物对厄洛替尼耐药。此外，在厄洛替尼耐药细胞中，PHGDH缺失特异性地改变了多个DNA损伤修复途径相关基因。SiPHGDH和NCT-503可增强ROS胁迫和DNA损伤标志物γ H2 AX的表达，NAC可逆转上述作用。结论：我们的研究表明，PHGDH抑制剂对肺腺癌具有潜在的治疗价值，并获得性耐药EGFR-TKI。
How to improve the efficacy and reverse the resistance to epidermal growth factor receptor tyrosine kinase inhibitors (EGFR-TKIs), such as erlotinib, remains a major challenge in the targeted therapy of lung adenocarcinoma with EGFR-activating mutation. Phosphoglycerate dehydrogenase (PHGDH) is the key enzyme of de novo serine biosynthesis over-expressed in various types of cancer including lung cancer. Elevated PHGDH expression is correlated with a worse overall survival in clinical lung adenocarcinoma patients. Here we investigated the role of PHGDH in lung adenocarcinoma with the acquisition of resistance to erlotinib. Methods: The necessary genes required for the acquired erlotinib resistance in lung adenocarcinoma cells were screened out by RNA-Seq analysis. Then the protein and mRNA levels of PHGDH were confirmed by immunoblotting and qRT-PCR in the erlotinib resistant cells. The effects of PHGDH inhibition or overexpression on erlotinib resistance were examined using cell culture and tumor xenograft mouse models respectively. To explore mechanism, the ROS level and DNA damage marker, γH2AX, were tested by DCFH-DA staining and immunofluorescence after PHGDH inhibition. Results: We found that PHGDH level was significantly increased in the lung adenocarcinoma PC9ER4 and HCC827ER9 cells that acquired resistance to erlotinib. Perturbation of PHGDH by siPHGDH transfection or NCT-503, a small molecular PHGDH inhibitor, synergistically augmented the tumoricidal effect and restored sensitivity to erlotinib in cell lines and xenografts. Over-expression of PHGDH caused xenografts resistant to erlotinib. Furthermore, multiple DNA damage repair pathways related genes were changed by PHGDH depletion specifically in erlotinib resistant cells. ROS stress and DNA damage marker γH2AX were enhanced by siPHGDH and NCT-503, which was reversed by NAC. Conclusion: Our study indicated that PHGDH inhibition has potential therapeutic value in lung adenocarcinoma with the acquired resistance to EGFR-TKIs.
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影响因子：
14.8
作者：
Pacold ME;Brimacombe KR;Chan SH;Rohde JM;Lewis CA;Swier LJ;Possemato R;Chen WW;Sullivan LB;Fiske BP;Cho S;Freinkman E;Birsoy K;Abu-Remaileh M;Shaul YD;Liu CM;Zhou M;Koh MJ;Chung H;Davidson SM;Luengo A;Wang AQ;Xu X;Yasgar A;Liu L;Rai G;Westover KD;Vander Heiden MG;Shen M;Gray NS;Boxer MB;Sabatini DM
通讯作者：
Sabatini DM
影响因子：
4
作者：
Jotte RM;Spigel DR
通讯作者：
Spigel DR
影响因子：
--
作者：
TeSlaa, Tara;Teitell, Michael A.
通讯作者：
Teitell, Michael A.
影响因子：
64.8
作者：
Possemato, Richard;Marks, Kevin M.;Shaul, Yoav D.;Pacold, Michael E.;Kim, Dohoon;Birsoy, Kivanc;Sethumadhavan, Shalini;Woo, Hin-Koon;Jang, Hyun G.;Jha, Abhishek K.;Chen, Walter W.;Barrett, Francesca G.;Stransky, Nicolas;Tsun, Zhi-Yang;Cowley, Glenn S.;Barretina, Jordi;Kalaany, Nada Y.;Hsu, Peggy P.;Ottina, Kathleen;Chan, Albert M.;Yuan, Bingbing;Garraway, Levi A.;Root, David E.;Mino-Kenudson, Mari;Brachtel, Elena F.;Driggers, Edward M.;Sabatini, David M.
通讯作者：
Sabatini, David M.
影响因子：
8.6
作者：
Wang Q;Liberti MV;Liu P;Deng X;Liu Y;Locasale JW;Lai L
通讯作者：
Lai L