贝伐单抗对人眼脉络膜黑色素瘤MUM-2B细胞血管生成拟态的影响  

The effect of bevacizumab on the angiogenesis mimicry of human choroidal melanoma MUM-2B cells

作　　者：周钰娟 丁瑞麟 彭红菊 李源 倪来超 文庆莲 ZHOU Yujuan;DING Ruilin;PENG Hongju;LI Yuan;NI Laichao;WEN Qinglian(Health Management Center,West China Hospital of Sichuan University,Sichuan Chengdu 610000,China;GCP Center,Affiliated Hospital of Southwest Medical University,Sichuan Luzhou 646000,China;Department of Oncology,the Second People's Hospital of Neijiang,Sichuan Neijiang 641000,China;Department of Oncology,Langzhong People's Hospital,Sichuan Langzhong 637400,China;Department of Oncology,Ya'an People's Hospital,Sichuan Ya'an 625000,China;Department of Oncology,Affiliated Hospital of Southwest Medical University,Sichuan Luzhou 646000,China)

机构地区：[1]四川大学华西医院健康管理中心,四川成都610000 [2]西南医科大学附属医院GCP中心,四川泸州646000 [3]内江市第二人民医院肿瘤科,四川内江641000 [4]阆中市人民医院肿瘤科,四川阆中637400 [5]雅安市人民医院肿瘤科,四川雅安625000 [6]西南医科大学附属医院肿瘤科,四川泸州646000

出　　处：《现代肿瘤医学》2023年第2期197-202,共6页Journal of Modern Oncology

基　　金：北京医卫健康公益基金会(编号:HR20002)。

摘　　要：目的:通过体内外实验探讨贝伐单抗对人眼脉络膜黑色素瘤细胞(MUM-2B)血管生成拟态的影响及可能相关的分子通路。方法:通过不同浓度的贝伐单抗(0,0.1,1,2,3 mg/mL)处理MUM-2B细胞,观察其对MUM-2B细胞成管能力的影响。通过CCK-8法及Transwell法检测其对MUM-2B细胞增殖及侵袭的影响。然后,建立裸鼠皮下移植瘤模型,通过不同剂量的贝伐单抗(5 mg/kg,7.5 mg/kg,10 mg/kg)干预,探究贝伐单抗对裸鼠皮下移植瘤VM形成的影响,并通过CD31/PAS免疫组化双重染色法检测VM的表达情况,免疫组化法检测HIF-1a、VE-cadherin、EphA2、PI3K蛋白表达水平变化。结果:体外实验中,不同浓度的贝伐单抗(0,0.1,1,2,3 mg/mL)处理MUM-2B细胞24小时后,对MUM-2B细胞的成管能力、增殖、侵袭能力并没有表现出明显的抑制或促进作用。实验组与对照组相比差异无统计学意义(P>0.05)。在体内实验中,贝伐单抗实验组(5 mg/kg,7.5 mg/kg,10 mg/kg)与对照组相比较能显著的促进MUM-2B细胞形成VM的能力(P<0.05),且形成管道的数量与贝伐单抗的浓度呈正相关(r=0.942,P<0.05)。实验组HIF-1a、VE-cadherin、EphA2和PI3K-Akt蛋白的表达水平均明显高于对照组(P<0.05),且各分子的表达量随着贝伐单抗浓度的升高呈浓度依赖性的升高。结论:体外实验中,贝伐单抗对MUM-2B细胞的成管能力,增殖,侵袭能力并没有表现出明显的抑制或促进作用。体内实验中,贝伐单抗的使用能促进HIF-1a的表达、上调VE-cadherin/EphA2/PI3K-Akt后续的级连信号通路,从而加速VM的生成。Objective:To investigate the effect of bevacizumab on the angiogenic mimicry of human choroidal melanoma cell(MUM-2 B)and the possible related molecular pathways in vitro and in vivo.Methods:MUM-2 B cells were treated with different concentrations of bevacizumab(0,0.1,1,2,3 mg/mL)to observe its effect on the tube-forming ability of MUM-2 B cells.The effects of the proliferation and invasion of MUM-2 B cells were detected by CCK-8 and Transwell assay.Then,the subcutaneous transplantation tumor models were established in nude mice,and the effect of bevacizumab on VM formation of subcutaneous transplantation tumors in it was investigated by different doses of bevacizumab(5 mg/kg,7.5 mg/kg,10 mg/kg)intervention,and the expression of VM was detected by double staining with CD31/PAS immunohistochemistry.The changes in the expression levels of HIF-1 a,VE-cadherin,EphA2,and PI3 K proteins were detected by immunohistochemistry.Results:In vitro,different concentrations of bevacizumab(0,0.1,1,2,3 mg/mL)treated with MUM-2 B cells for 24 hours did not show significant inhibitory or promotional effects on the tube-forming ability,proliferation,and invasion ability of MUM-2 B cells.The difference between the experimental group and the control group was not statistically significant(P>0.05).In vivo,compared with the control group,the bevacizumab experimental group(5 mg/kg,7.5 mg/kg,10 mg/kg)could significantly promote the ability of MUM-2 B cells to form VM(P<0.05),and the number of tubes was positively correlated with the concentration of bevacizumab(r=0.942,P<0.05).The expression levels of HIF-1 a,VE-cadherin,EphA2,and PI3 K-Akt proteins were significantly higher in the experimental group than in the control group(P<0.05),and the expression of each molecule increased in a concentration-dependently manner with the increase of bevacizumab concentration.Conclusion:In vitro,bevacizumab did not significantly inhibit or promote the tubulation,proliferation and invasion of MUM-2 B cells.In vivo,Bevacizumab can promote the expression

关 键 词：贝伐单抗 人眼脉络膜黑色素瘤 血管生成拟态 增殖 侵袭 HIF-1a/VE-cadherin/EphA2/PI3K-Akt 

分 类 号：R739.5[医药卫生—肿瘤]