CAJ | 학술논문

目的观察吉非替尼对人表皮生长因子受体(EGFR)高表达的三阴性乳腺癌(TNBC)细胞系MDA-MB-231迁移能力的影响.方法采用Western blot法检测不同浓度吉非替尼作用MDAMB-231细胞后EGFR和Akt磷酸化水平；采用划痕实验和Boyden小室趋化实验观察吉非替尼对细胞迁移、趋化能力的影响；采用免疫荧光染色观察吉非替尼对细胞骨架重构及极性变化的影响.结果与对照组(0 μmol/L吉非替尼)相比,不同浓度的吉非替尼可有效抑制EGFR及其下游通路关键蛋白的磷酸化水平,呈明显的量效关系.划痕实验24h后,0、0.1、1、10、20 μmol/L吉非替尼组细胞迁移距离分别为(36.3 ±4.0)μm、(30.3±3.8)μm、(26.8±3.3) μm、(17.0±2.6) μm和(11.0 ±2.5)μm；Boyden小室趋化实验3.5h后,0、0.1、1、10、20 μmol/L吉非替尼组的穿膜细胞数分别为(69.2±7.0)个、(51.8±7.5)个、(43.8±8.7)个、(30.6±4.8)个和(28.4±3.4)个.吉非替尼可明显延长划痕愈合时间,减少趋化穿膜细胞个数,差异有统计学意义(P＜0.05).吉非替尼可明显减少细胞片状伪足的形成,抑制细胞骨架的重构及极性变化.结论吉非替尼可以通过抑制TNBC细胞系MDA-MB-231中EGFR/PI3 K/Akt通路的磷酸化水平,抑制细胞骨架蛋白(微丝)的重构及极性改变,从而降低细胞的迁移运动能力.
목적 관찰길비체니대인표피생장인자수체(EGFR)고표체적삼음성유선암(TNBC)세포계MDA-MB-231천이능력적영향.방법 채용Western blot법검측불동농도길비체니작용MDAMB-231세포후EGFR화Akt린산화수평；채용화흔실험화Boyden소실추화실험관찰길비체니대세포천이、추화능력적영향；채용면역형광염색관찰길비체니대세포골가중구급겁성변화적영향.결과 여대조조(0 μmol/L길비체니)상비,불동농도적길비체니가유효억제EGFR급기하유통로관건단백적린산화수평,정명현적량효관계.화흔실험24h후,0、0.1、1、10、20 μmol/L길비체니조세포천이거리분별위(36.3 ±4.0)μm、(30.3±3.8)μm、(26.8±3.3) μm、(17.0±2.6) μm화(11.0 ±2.5)μm；Boyden소실추화실험3.5h후,0、0.1、1、10、20 μmol/L길비체니조적천막세포수분별위(69.2±7.0)개、(51.8±7.5)개、(43.8±8.7)개、(30.6±4.8)개화(28.4±3.4)개.길비체니가명현연장화흔유합시간,감소추화천막세포개수,차이유통계학의의(P＜0.05).길비체니가명현감소세포편상위족적형성,억제세포골가적중구급겁성변화.결론 길비체니가이통과억제TNBC세포계MDA-MB-231중EGFR/PI3 K/Akt통로적린산화수평,억제세포골가단백(미사)적중구급겁성개변,종이강저세포적천이운동능력.
Objective To investigate the effect of gefitinib on the migration of triple-negative breast cancer cell line MDA-MB-231 cells.Methods Gefitinib was used in concentrations of 0 μnol/L,0.1 μmol/L,1 μmol/L,10 μmol/L and 20 μmol/L,respectively.Phosphorylation levels of EGFR and Akt were analyzed by Western blot.The capability of migration was measured by scratch test and Boyden chamber assay. Microfilaments (cell skeleton ) remolding and polarization were evaluated by immunofluorescence microscopy. Results Comparing with the control group (0 μmol/L gefitinib ),gefitinib effectively inhibited the phosphorylation of EGFR and its downstream key proteins,and the effect displayed an obvious dose-effect relationship.At 24 hours after wound scratch,the cell migration distance of each group with 0,0.1,1,10,20 μmol/L gefitinib was (36.3 ± 4.0) μm,( 30.3 ± 3.8 ) μm,( 26.8 ±3.3)μm,(17.0 ±2.6) μm,and ( 11.0 ±2.5) μm,respectively.At 3.5 hours after Boyden chamber assay,the cell count of each group with 0,0.1,1,10,20 μnol/L gefitinib was 69.2 ± 7.0,51.8 ± 7.5,43.8 ± 8.7,30.6 ± 4.8,and 28.4 ± 3.4,respectively.Compared with the control group (0 μmol/L gefitinib),gefitinib could significantly prolong the wound-healing time and decrease the migrating cell count (P ＜0.05),and significantly inhibit the lamellipodium formation,cell skeleton remolding and changes of the cytoskeleton polarization.Conclusions Gefitinib can reduce the migration capacity of triple-negative breast cancer cells through inhibiting phosphorylation of EGFR/PI3K/Akt pathway,suppressing the cell skeleton (microfilaments) remolding and changes of its polarization.