新型VEGF单克隆抗体与全反式维甲酸联合应用对乳腺癌MCF-7细胞增殖的抑制作用  

Inhibitory effect of new VEGF monoclonal antibody combined with all-trans retinoic acid on proliferation of breast cancer MCF-7 cells

作　　者：唐浩轩 孟楠峰 杜美玲 王伟[1] 何涛[1,2] TANG Haoxuan;MENG Nanfeng;DU Meiling;WANG Wei;HE Tao(Graduate School of Baotou Medical College,Inner Mongolia University of Science and Technology,Baotou 014010,China;Department of General Surgery,First Affiliated Hospital,Baotou Medical College,Inner Mongolia University of Science and Technology,Baotou 014010,China)

机构地区：[1]内蒙古科技大学包头医学院研究生学院,内蒙古包头014010 [2]内蒙古科技大学包头医学院第一附属医院普外科,内蒙古包头014010

出　　处：《吉林大学学报（医学版）》2021年第2期344-351,共8页Journal of Jilin University：Medicine Edition

基　　金：内蒙古自治区科技厅科技计划项目(201702101);包头医学院科学研究基金项目(BYJJ-YF-2018003)。

摘　　要：目的:探讨新型血管内皮生长因子(VEGF)单克隆抗体、全反式维甲酸(ATRA)、新型VEGF单克隆抗体与ATRA联合用药对乳腺癌MCF-7细胞的影响,为乳腺癌的临床治疗提供新方案。方法:将MCF-7乳腺癌细胞分为对照组(不做任何处理)、VEGF抗体组(20 mg·L~(-1)VEGF抗体)、ATRA组(10μmol·L~(-1)ATRA)和VEGF抗体+ATRA组(20 mg·L~(-1)VEGF抗体+10μmol·L~(-1)ATRA)4组,每组5个重复。加入不同药物处理MCF-7细胞48 h后,采用ELISA法检测新型VEGF抗体浓度,MTT法检测MCF-7细胞增殖率,流式细胞术检测MCF-7细胞凋亡率,免疫印迹法检测MCF-7细胞中B细胞淋巴瘤2(Bcl-2)、Bcl-2相关X蛋白(Bax)和核转录因子κB(NF-κB)蛋白表达水平。结果:ELISA法检测,新型人类重组VEGF单克隆抗体浓度为200μg·L~(-1),与阳性抗体对照比较其质量良好。MTT法检测,与对照组比较,VEGF抗体+ATRA组MCF-7细胞增殖率明显降低(P<0.01)。流式细胞术检测,与对照组、VEGF抗体组和ATRA组比较,VEGF抗体+ATRA组MCF-7细胞凋亡率明显升高(P<0.01);与对照组比较,VEGF抗体组和ATRA组MCF-7细胞凋亡率明显升高(P<0.01);与VEGF抗体组比较,ATRA组MCF-7细胞凋亡率升高(P<0.01)。免疫印迹法检测,与对照组、VEGF抗体组和ATRA组比较,VEGF抗体+ATRA组MCF-7细胞中Bcl-2蛋白表达水平降低(P<0.05);与对照组和VEGF抗体组比较,ATRA组MCF细胞中NF-κB蛋白表达水平明显降低(P<0.05或P<0.01),Bax蛋白表达水平明显升高(P<0.01)。结论:新型VEGF单克隆抗体和ATRA可以抑制乳腺癌MCF-7细胞增殖、促进细胞凋亡,而二者联合用药的效果要显著优于单独用药。Objective:To explore the effects of new vascular endothelial growth factor(VEGF)monoclonal antibody,all-trans retinoic acid(ATRA),new VEGF monoclonal antibody combined with ATRA on the breast cancer MCF-7 cells,and to provide a new clinical approach for the treatment of breast cancer.Methods:The breast cancer MCF-7 cells were divided into control group(without any treatment),VEGF antibody group(20 mg·L-1 VEGF antibody),ATRA group(10μmol·L-1 ATRA)and VEGF antibody+ATRA group(20 mg·L-1 VEGF antibody+10μmol·L-1 ATRA),each with5 replicates.After the MCF-7 cells were treated with different drugs for 48 h,the VEGF antibody concentration was determined by ELISA,the proliferation rate of MCF-7 cells was determined by MTT assay,the apoptotic rate of MCF-7 cells was determined by flow cytometry,and the expression levels of B cell lymphoma-2(Bcl-2),Bcl-2 associated X protein(Bax)and nuclear factor-κB(NF-κB)proteins in the MCF-7 cells were determined by Western blotting method.Results:The ELISA results showed that the new human recombinant VEGF monoclonal antibody had good quality compared with positive antibody control and its concentration was 200μg·L-1.The MTT results showed that the prolifenation rate of MCF-7 cells in VEGF antibody+ATRA group was decreased obviously compared with control group(P<0.01).The flow cytometry results showed that compared with control group,VEGF antibody group and ATRA group,the apoptotic rate of MCF-7 cells in VEGF antibody+ATRA group was significantly increased(P<0.01);compared with control group,the apoptotic rates of MCF-7 cells in VEGF antibody group and ATRA group were significantly increased(P<0.01);compared with VEGF antibody group,the apototic rate of MCF-7 cells in ATRA group was significantly increased(P<0.01).The results of Western blotting method showed that compared with control group,VEGF antibody group and ATRA group,the expression level of Bcl-2 protein in VEGF antibody+ATRA group was significantly decreased(P<0.05);compared with control group and VEGF antibody group

关 键 词：乳腺肿瘤 血管内皮生长因子 单克隆抗体 全反式维甲酸 细胞凋亡 细胞增殖 

分 类 号：R737.9[医药卫生—肿瘤]