摘要
目的 观察偶联西妥昔单抗(C225)的金纳米颗粒(AuNPs)对肺腺癌A549细胞株的作用,研究其对细胞功能以及相关蛋白的影响并探讨其分子机制.方法 应用浓度分别为1、10、100、500 mg/L的C225、偶联IgG的金纳米颗粒(IgG-AuNPs)、偶联C225的金纳米颗粒(C225-AuNPs)处理A549细胞,药物作用24、48 h后通过细胞计数试剂盒(CCK-8)检测细胞抑制率;通过流式细胞仪检测细胞凋亡和周期变化,通过Transwell小室法检测细胞迁移,Western blot检测表皮生长因子受体(EGFR)、蛋白激酶B(Akt)、磷酸化Akt (p-Akt)、细胞外信号调节激酶(ERK)、磷酸化ERK(p-ERK)以及B细胞淋巴瘤/白血病-2(bcl-2)蛋白的表达变化.结果 C225-AuNPs对A549细胞株的抑制率呈剂量依赖性,当浓度达到100 mg/L时,最大抑制率为28%,相对单用IgG-AuNPs和C225,C225-AuNPs显著抑制细胞增殖.100 mg/L的C225-AuNPs作用后A549细胞凋亡率为(27.40±3.54)%,明显高于相同浓度C225的(11.80±2.02)%以及IgG-AuNPs的(9.50±2.76)%;但是C225-AuNPs对细胞周期的影响并不明显;C225-AuNPs处理组后穿膜细胞数目为(94±9)个,与C225组[(155±12)个]以及IgG-AuNPs组[(136±15)个]比较明显减少;通过Western blot检测经过C225-AuNPs处理后p-AKt、p-ERK以及bcl-2蛋白的表达量较C225以及IgG-AuNPs组明显下调.结论 C225-AuNPs显著抑制A549细胞增殖、迁移,促进凋亡,增强A549细胞对C225单抗的敏感性.
Objective To investigate whether a specific nanoprobe of epidermal growth factor receptor (EGFR) which conjugating cetuximab to gold nanoparticles can affect physiological activities of A549 cells,and the molecular mechanism.Methods A549 cells were treated with different concentrations of cetuximab (C225),IgG-gold nanoparticles (AuNPs) and C225-AuNPs (1,10,100,500 mg/L).Cell survival was detected by cell counting kit-8 (CCK-8) assay.Apoptotic cells were quantitatively examined using FITC Annexin V apoptosis Detection Kit I.Cell cycle was tested by flow cytometry.The invasion ability was measured through the Transwell method.The protein expression of EGFR,protein kinase B (Akt),p-Akt,extracellular signal-regulated kinase (ERK),p-ERK and B lymphocytes/leukemia-2 (bcl-2) was detected by using Western blotting.Results The cytotoxicity of C225-AuNPs on the lung cancer A549 cells was increased in a dose-dependent manner.C225-AuNPs suppressed proliferation significantly.100 mg/L C225-AuNPs,C225 and IgG-AuNPs increased apoptosis rate to (27.40 ± 3.54) %,(11.80 ± 2.02)% and (9.50 ±2.76)%,respectively.A549 cells treated with 100 mg/L C225-AuNPs had fewer cells penetrating through the membrane than the control groups [C225-AuNPs mean:(94 ± 9) vs.C225 group mean:(155 ± 12) vs.IgG-AuNPs group mean:(136 ± 15)].The expression of p-AKt,p-Erk and bcl-2 was reduced significantly through C225-AuNPs treatments.Conclusion C225-AuNPs suppressed proliferation and invasion,accelerated apoptosis of A549 cells,and increased the sensibility to C225.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2014年第3期580-583,共4页
Chinese Journal of Experimental Surgery
基金
卫生部基金资助项目(W6)
江苏省自然科学重点研究专项基金资助项目(BK2011036)
江苏省科技厅临床科技重点专项基金资助项目(BL2012030)
关键词
西妥昔单抗
金纳米
表皮生长因子受体
靶向治疗
非小细胞肺癌
Cetuximab
Gold nanoparticles
Epidermal growth factor receptor
Targeted therapy
Non-small cell lung cancer