摘要
目的:探讨碱性成纤维细胞生长因子单克隆抗体(monoclonal antibody to basic broblast growth factor,bFGF mAb)通过P-糖蛋白(permeability glycoprotein,P-gp)对人乳腺癌多柔比星(adriamycin,ADM)耐药细胞株MCF-7/ADM多药耐药(multidrug resistance,MDR)性的逆转机制。方法:CCK-8(cell counting kit-8)法检测bFGF mAb对MCF-7/ADM细胞增殖的影响及其对化疗药物耐药的逆转作用;bFGF mAb作用后,FCM检测MCF-7/ADM细胞的细胞周期、P-gp的表达及细胞内罗丹明(rhodamine,Rho)123的荧光强度,实时荧光定量PCR检测MCF-7/ADM细胞中MDR1和碱性成纤维细胞生长因子(basic broblast growth factor,bFGF)mRNA的表达。结果:1μg/mL bFGF mAb对MCF-7和MCF-7/ADM细胞的抑制率分别为(19.87±1.05)%和(27.34±2.79)%,差异有统计学意义(P<0.01)。bFGF mAb可有效逆转MCF-7/ADM细胞对ADM、吉西他滨(gemcitabine,GEM)和奥沙利泊(oxaliplatin,OXA)的耐药性,逆转指数分别为4.46、4.25和2.18。bFGF mAb作用MCF-7/ADM细胞后,细胞阻滞于G0/G1期,P-gp表达下调,细胞内Rho123的荧光强度增强,MDR1和bFGF mRNA的表达下调,与未作用组比较,差异有统计学意义(P<0.01)。结论:bFGF mAb能抑制MCF-7/ADM细胞增殖并逆转其MDR,该作用机制可能与bFGF mAb下调MDR1/P-gp表达、抑制P-gp功能以及增加细胞内化疗药物浓度有关。
Objective： To investigate the mechanism of reversal effect of bFGF mAb （monoclonal antibody to basic fibroblast growth factor）-mediated expression of P-gp （permeability glycoprotein） on MDR （multidrug resistance） in ADM （adriamycin）-resistant human breast cancer cell line MCF-7/ADM. Methods： The effects of bFGF mAb on the proliferation of MCF-7/ADM cells and the reversal of MDR were detected by CCK-8 （cell counting kit-8） method. The cell cycle distribution of MCF-7/ADM cells and the expression of P-gp and intracellular fluorescence intensity of Rho123 （rhodamine 123） in MCF-7/ADM cells after bFGF mAb intervention were analyzed by flow cytometry. The expressions of MDR 1 （multidrug resistance protein 1） and bFGF mRNAs in MCF-7/ADM cells were examined by real-time fluorescence quantitative PCR. Results： The growth inhibition rates of MCF-7 cells and MCF-7/ADM cells after treatment with 1 μg/mLbFGF mAb were （19.87±1.05）% and （27.34±2.79）% （P 〈 0.01）. bFGF mAb intervention could reverse ADM-, GEM （gemcitabine）- and OXA （oxaliplatin）-resistance of MCF-7/ADM cells, and the corresponding reversal index were 4.46, 4.25 and 2.1 8, respectively. As compared with the MCF-7/ADM cells without bFGF mAb intervention, the cell cycle of MCF-7/ADM cells after bFGF mAb intervention was arrested at G0/G1 phase, the expression level of P-gp was down-regulated, the intracellular Rho123 fluorescence intensity was increased, and the expression levels of MDR 1 and bFGF mRNAs were both decreased （P 〈 0.01）. Conclusion： bFGF mAb can inhibit the proliferation of MCF-7/ADM and reverse MDR. This mechanism may be related to down-regulation of the expression levels of MDR 1 and P-gp, inhibition of the function of P-gp, and increasement of intracellular concentration of chemotherapeutic drugs.
出处
《肿瘤》
CAS
CSCD
北大核心
2013年第1期8-14,共7页
Tumor
基金
国家高技术研究发展计划(863计划)资助项目(编号:2009AA02Z112)
国家自然科学基金资助项目(编号:81273814)
关键词
乳腺肿瘤
碱性成纤维细胞生长因子
抗体
单克隆
P-糖蛋白
抗药性
肿瘤
Breast neoplasms
Basic fibroblast growth factor
Antibody, monoclonal
P-glycoprotein
Drug resistance, neoplasm
