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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 study the antitumor effect of DCLAK11 on non-small cell lung cancer (NSCLC) and its mechanism. Methods: The effects of DCLAK11 on the proliferation of HCC827 cells were detected by SRB assay and clone formation assay. The effect of DCLAK11 on the apoptosis of HCC827 cells was detected by flow cytometry. The effect of DCLAK11 on the migration of human umbilical vein endothelial cells HUVEC was detected by wound healing assay. The effect of DCLAK11 on the epidermal growth factor receptor (EGFR) phosphorylation of HCC827 cells was detected by immunofluorescence assay. The ATP competition experiment was conducted to investigate whether DCLAK11 has ATP-competitive properties on EGFR kinase activity. RESULTS: DCLAK11 significantly inhibited the cell viability (P<0.01) and colony formation of HCC827 cells; DCLAK11 induced apoptosis of HCC827 cells; DCLAK11 inhibited the wound healing of HUVECs in a concentration- and time-dependent manner (P<0.01). DCLAK11 inhibited the phosphorylation of EGFR in HCC827 cells; DCLAK11 inhibited EGFR kinase activity by ATP competition. CONCLUSION: DCLAK11 is an ATP-competitive small molecule kinase inhibitor that can significantly inhibit the proliferation of HCC827 cells and induce apoptosis in NSCLC cells. These effects may be related to the inhibition of EGFR activation by DCLAK11. In addition, DCLAK11 also has the effect of inhibiting angiogenesis.