抗血管生成药物增加血清饥饿环境中MDA—MB-231乳腺癌细胞的侵袭和迁移能力  

Antiangiogenic drugs enhance the ability of invasion and metastasis of breast cancer cells under serum starvation and hypoxia

作　　者：刘薇[1] 张翠翠[1] 王晶[1] 李凯[1] 

机构地区：[1]天津医科大学肿瘤医院肺部肿瘤内科天津市肿瘤防治重点实验室国家肿瘤临床医学研究中心,300060

出　　处：《中华肿瘤杂志》2015年第4期244-250,共7页Chinese Journal of Oncology

基　　金：天津市抗癌重大专项攻关计划（12ZCDSY15600）;天津市自然科学基金项目（11JCYBJC11300）

摘　　要：目的观察在缺氧和（或）血清饥饿环境中，重组人血管内皮抑素和贝伐珠单抗对人乳腺癌MDA—MB-231细胞侵袭和迁移能力的影响，探讨抗血管生成药物治疗的潜在风险。方法将MDA—MB-231细胞分别置于4种环境培养，分别为营养环境（10％胎牛血清）、缺氧（1％氧气）＋营养环境、饥饿环境（无胎牛血清）和缺氧＋饥饿环境。在每种培养环境中，分别设空白组（未给予抗血管生成药物）、抑素组（给予重组人血管内皮抑素）和单抗组（给予贝伐珠单抗）。以CCK-8法计算重组人血管内皮抑素及贝伐珠单抗的细胞生长抑制率，确定其最适工作浓度与时间，并对各组细胞行Transwell细胞侵袭实验和细胞迁移实验，以Western blot检测细胞中c—Met和基质金属蛋白酶9（MMP-9）的表达。对饥饿环境中的处理组细胞行ExiqonmiRBase18．0microRNA芯片杂交，以实时荧光定量PCR法验证杂交芯片实验中检测到的微小RNA。结果800mg／L重组人血管内皮抑素作用48h和24h后，细胞抑制率分别为（32．2±2．5）％和（27．0±1．3）％（P=0．023）；80mg／L贝伐珠单抗作用48h和24h后，细胞抑制率分别为（30．5±1．4）％和（26．1±2．4）％（P=0．015）。在饥饿环境中，空白组侵袭、迁移穿膜细胞数分别为（28．8±2．2）个和（31．4±1．5）个，与单抗组和抑素组比较，差异均有统计学意义（均P〈0．05）。空白组中c-Met和MMP-9蛋白的相对表达量分别为0．213±0．017和0．542±0．048，与单抗组和抑素组比较，差异均有统计学意义（均P〈0．05）。在缺氧环境中，抑素组侵袭、迁移穿膜细胞数分别为（17．5±2．1）个和（16．5±2．8）个，单抗组分别为（16．3±3．5）个和（17．5±2．4）个，与空白组比较，差异均无统计学意义（均P〉0．05）。缺氧对细胞侵袭迁移能力、c—Met和MMP-9的表达亦无影响，差异无�Objective To investigate the ability of invasion and migration of breast cancer MDA- MB-231 cells under serum starvation and hypoxia, and the effect of antiangiogenic drugs, rh-endostatin and bevacizumab, on the ability of invasion and migration of breast cancer cells under serum starvation and/or hypoxia, in order to explore the potential risk of antiangiogenic therapy in clinics. Methods The cells were randomized into 4 groups, i.e., group A： 10% fetal bovine serum （FBS） group; group B： hypoxia ＋ 10% FBS group; group C： serum starvation group; group D： hypoxia ＋ serum starvation group; each group was further divided into three subgroups as blank control, treated with rh-endostatin and bevacizumab, respectively. Cell counting kit-8 （CCK-8） was used to assess the inhibition rate of cell growth induced by endostatin and bevacizumab, in order to determine the proper working concentration and time of the two drugs. Transwell assay was conducted to detect the cell invasion and migration in vitro. The expressions of c- Met and MMP-9 were detected by Western blot. The cells treated with rh-endostatin or bevacizumab under serum starvation were tested by hybridization using Exiqon miBase 18.0 microarray. The miRNAs which exibited significant differences （P〈0.05） in miRNA hybridization were verified by real-time PCR assay. Results CCK-8 assay showed that the inhibition rates of MDA-MB-231 cells cultured with 800 mg/L rh- endostatin for 48 h and 24 h were （ 32.2 ± 2.5 ） % and （ 27.0 ± 1.3 ） %, respectively, showing a significant difference （P= 0.023）. The inhibition rates of MDA-MB-231 cells cultured with 80 mg/L bevacizumab for 48 h and 24 h were （30.5±1.4）% and （26.1±2.4）%, respectively , showing also a significant difference （P=0.015）. The Transwell assay showed that in the starvation blank group, the number of invaded and penetrated cells were 28.8 ± 2.2 and 31.4 ± 1.5, respectively, significantly different from that in the rhendostatin and bevacizumab groups （P

关 键 词：乳腺肿瘤 血管生成抑制剂 胎牛血清 缺氧 肿瘤浸润 

分 类 号：R737.9[医药卫生—肿瘤]