小凹蛋白-1调控表皮生长因子受体活化对恶性黑色素瘤细胞生物学行为的影响  

Effect of Caveolin-1 Regulating EGFR Activation on Biological Behavior of Malignant Melanoma Cells

作　　者：张玫莹 朱铁年[2] 赵瑞景[3] 崔玉洁[2] 范婧 ZHANG Mei-ying;ZHU Tie-nian;ZHAO Rui-jing;CUI Yu-jie;FAN Jing(Chengde Medical College,Chengde,Hebei 067000,China;Department of Oncology,Bethune International Peace Hospital of PLA,Shijiazhuang 050082,China;Department of Immunology,Hebei Medical University,Shijiazhuang 050017,China)

机构地区：[1]承德医学院,河北承德067000 [2]解放军白求恩国际和平医院肿瘤科,石家庄050082 [3]河北医科大学免疫教研室,石家庄050017

出　　处：《解放军医药杂志》2018年第4期46-50,共5页Medical & Pharmaceutical Journal of Chinese People’s Liberation Army

基　　金：国家自然科学基金项目(81071846);河北省科技厅国际合作项目(14397707D;12396107D);吴阶平基金项目(320.6750.12604;320.6750.14063;320.6799.15005)

摘　　要：目的探讨小凹蛋白-1(Cav-1)调控表皮生长因子受体(EGFR)活化对恶性黑色素瘤细胞生物学行为的影响。方法体外培养恶性黑色素瘤M2细胞,利用质粒转染技术建立过表达Cav-1的稳定细胞株M2(Cav-1)并验证;MTS法、细胞划痕实验以及Transwell侵袭实验分别检测稳定转染细胞株的增殖、迁移与侵袭能力;蛋白免疫印迹法检测稳转细胞株EGFR及其下游信号转导通路的活化水平。结果通过质粒转染技术得到的M2(Cav-1)细胞中Cav-1的表达量明显高于M2(pcDNA3.1)细胞(P<0.01)。在不同浓度表皮生长因子(EGF)刺激下,M2(Cav-1)细胞的增殖能力、迁移能力、侵袭能力均低于M2(pcDNA3.1)细胞(P<0.01)。M2(Cav-1)细胞经EGF刺激后磷酸化EGFR、ERK及Akt水平明显低于M2(pcDNA3.1)细胞(P<0.01)。结论 Cav-1可通过调控恶性黑色素瘤M2细胞EGFR的活化水平,影响MAPK/ERK及PI3K/Akt信号转导通路激酶的活化,进而影响M2细胞的生物学行为。Objective To investigate effect of caveolin-1(Cav-1)regulating epidermal growth factor receptor(EGFR)activation on biological behaviour of malignant melanoma cells.Methods Malignant melanoma M2 cells were cultured in vitro,and stable M2(Cav-1)cell line of Cav-1 overexpression was constructed by plasmid transfection technology,and then it was validated.MTS assay,cell scarification test and Transwell invasion test were used to respectively detect proliferation,migration and invasion abilities of stably transfected cell line.Western blotting was used to detect activation levels of stably transfected cell lines EGFR and its downstream signal transduction pathway.Results Cav-1 expression in M2(Cav-1)cell line obtained by plasmid transfection technology was significantly higher than that in M2(pcDNA3.1)cell(P<0.01).Proliferation,migration and invasion abilities of M2(Cav-1)cell by different concentration of epidermal growth factor(EGF)stimulation were significantly worse than those in M2(pcDNA3.1)cell(P<0.01).Levels of p-EGFR,p-ERK and p-Akt in M2(Cav-1)cells after EGF stimulation were significantly lower than those in M2(pcDNA3.1)cell(P<0.01).Conclusion Cav-1 may affect activation levels of MAPK/ERK and PI3K/Akt signal transduction pathway kinase by regulating EGFR activation level in malignant melanoma M2 cells in order to affect biological behavior of M2 cells.

关 键 词：黑色素瘤 小凹蛋白-1 受体 表皮生长因子 细胞增殖 转移 

分 类 号：R739.5[医药卫生—肿瘤]