CAJ | 학술논문

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.
목적 관찰우련서타석단항(C225)적금납미과립(AuNPs)대폐선암A549세포주적작용,연구기대세포공능이급상관단백적영향병탐토기분자궤제.방법 응용농도분별위1、10、100、500 mg/L적C225、우련IgG적금납미과립(IgG-AuNPs)、우련C225적금납미과립(C225-AuNPs)처리A549세포,약물작용24、48 h후통과세포계수시제합(CCK-8)검측세포억제솔;통과류식세포의검측세포조망화주기변화,통과Transwell소실법검측세포천이,Western blot검측표피생장인자수체(EGFR)、단백격매B(Akt)、린산화Akt (p-Akt)、세포외신호조절격매(ERK)、린산화ERK(p-ERK)이급B세포림파류/백혈병-2(bcl-2)단백적표체변화.결과 C225-AuNPs대A549세포주적억제솔정제량의뢰성,당농도체도100 mg/L시,최대억제솔위28％,상대단용IgG-AuNPs화C225,C225-AuNPs현저억제세포증식.100 mg/L적C225-AuNPs작용후A549세포조망솔위(27.40±3.54)％,명현고우상동농도C225적(11.80±2.02)％이급IgG-AuNPs적(9.50±2.76)％;단시C225-AuNPs대세포주기적영향병불명현;C225-AuNPs처리조후천막세포수목위(94±9)개,여C225조[(155±12)개]이급IgG-AuNPs조[(136±15)개]비교명현감소;통과Western blot검측경과C225-AuNPs처리후p-AKt、p-ERK이급bcl-2단백적표체량교C225이급IgG-AuNPs조명현하조.결론 C225-AuNPs현저억제A549세포증식、천이,촉진조망,증강A549세포대C225단항적민감성.
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.