DCLAK11对非小细胞肺癌细胞HCC827的抗肿瘤作用

The anti-tumor effect of DCLAK11 against non-small cell lung cancer HCC827 cells

目的:研究DCLAK11对非小细胞肺癌(non-small cell lung cancer,NSCLC)的抗肿瘤作用及其机制。方法:SRB法和克隆形成实验检测DCLAK11对HCC827细胞增殖的影响;流式细胞术检测DCLAK11对HCC827细胞凋亡的影响;细胞划痕愈合实验检测DCLAK11对人脐静脉血内皮细胞HUVEC迁移的作用;免疫荧光实验检测DCLAK11对HCC827细胞表皮生长因子受体(epidermal growth factor receptor,EGFR)磷酸化的作用;ATP竞争实验考察DCLAK11对EGFR激酶活性是否具有ATP竞争性特性。结果:DCLAK11显著抑制NSCLC细胞HCC827的细胞活性(P<0.01)及克隆形成;DCLAK11可诱导HCC827细胞发生凋亡;DCLAK11浓度及时间依赖性地抑制血管内皮细胞HUVEC的划痕愈合(P<0.01);DCLAK11抑制HCC827细胞EGFR的磷酸化;DCLAK11对EGFR激酶活性的抑制作用具有ATP竞争性。结论:DCLAK11是ATP竞争性的小分子激酶抑制剂,可显著抑制NSCLC细胞HCC827的增殖及诱导该细胞发生凋亡,以上作用可能与DCLAK11抑制EGFR激活相关。此外,DCLAK11还具有抑制血管新生的作用。

OBJECTIVE To investigate the anti-cancer effects of DCLAK11 against non-small cell lung cancer （NSCLC） and its molecular mechanism. METHODS Cell viability was measured using SRB assay and colony-forming assay. Quantification of apoptosis was determined By flow cytometry. Wound healing assay was used to evaluate the impact on the migration of HU- VECs. Changes of epidermal growth factor receptor （EGFR） pbosphorylation in HCC827 were detected by immol/Lunofluo- rescence stain. ATP competitive inhibitory effect .of DCLAK11 on EGFR kinase was measured using ELISA assay. RESULTS DCLAK11 potently inhibited the proliferation of HCC827 （P〈0. 01）. DCLAKll effectively induced HCC827 cell apoptosis. The migration of HUVECs was down regulated in concentration and time dependent manner after dispose with DCLAK11 （P〈 0. 01 ）. DCLAK11 significantly inhibited phosphorylation of EGFR in HCC827. Inhibitory effect of DCLAK11 on EGFR kinase was ATP competitive. CONCLUSION DCLAK11 is an ATP competitive kinase inhibitor with remarkable potency against tyrosine kinase EGFR, which confirms its potent proliferation inhibiting and apoptosis promoting activity in HCC827 cells. Moreover, DCLAK11 exhibits anti-angiogenic activity.