CD147单克隆抗体对HCE1多细胞球体紫杉醇耐药的影响(英文)  

Effect of CD147 monoclonal antibody on paclitaxel resistance in HCE1 multicellular spheroids

作　　者：胡芸[1] 吴宜林[1] 

机构地区：[1]中南大学湘雅二医院妇产科,长沙410011

出　　处：《中南大学学报（医学版）》2011年第3期192-202,共11页Journal of Central South University ：Medical Science

基　　金：supported by the research fund from the Department of Education of Hunan Province,P.R.China(2050205)

摘　　要：目的:研究CD147单克隆抗体在人宫颈鳞癌细胞株HCE1多细胞球体模型中对紫杉醇天然耐药的影响,探讨CD147单克隆抗体是否可以逆转HCE1多细胞球体天然耐药及其对P-糖蛋白(P-gp)的影响。方法:运用液体重叠法和旋转法建立HCE1多细胞球体模型。以单层细胞作为对照,观察CD147单克隆抗体干预前后HCE1多细胞球体形态学变化。WST-1法检测不同浓度的CD147单克隆抗体干预前后,紫杉醇对HCE1多细胞球体的抑制率,计算半数抑制浓度(half maximal inhibitory concentration,IC50)和多细胞耐药指数(index of multicellular resistance,MCRI),并绘制浓度抑制率曲线。用对照组,CD147单抗,紫杉醇及紫杉醇+CD147单抗分别干预单层细胞及多细胞球体,流式细胞学检测细胞周期及凋亡率。用免疫组织化学法分别检测药物干预前后单层细胞及多细胞球体中CD147和P-gp的表达。结果:成功建立了HCE1多细胞球体紫杉醇天然耐药模型。CD147单克隆抗体能使HCE1多细胞球体解聚。CD147单克隆抗体能增强HCE1多细胞球体对紫杉醇的敏感性。5,10,20μg/mL CD147单克隆抗体组IC50分别为(40.31±3.73),(32.43±1.56),(30.69±1.01)μg/mL,5和10μg/mL CD147单抗组呈剂量依赖性(P<0.05)而10和20μg/mL CD147单抗组无明显剂量依赖性(P>0.05),其凋亡率接近单层细胞(P>0.05)。CD147单克隆抗体引起多细胞球体G1/G0期阻滞,和紫杉醇联用,分别在G1/G0和G2/M 2个调控点共同阻滞HCE1细胞生长。HCE1多细胞球体各组中CD147和p-gp表达呈一致性。结论:成功建立宫颈鳞癌HCE1多细胞球体紫杉醇天然耐药模型;CD147的表达与宫颈鳞癌HCE1多细胞球体对紫杉醇的耐药呈正相关,阻断CD147可部分逆转HCE1多细胞球体对紫杉醇的天然耐药;CD147介导的HCE1多细胞球体的紫杉醇天然耐药可能与P-gp有关。Objective To investigate the effect of CD147 monoclonal antibody（mAb） on the natural resistance to paclitaxel（TAX） in the human cervical cancer line（HCE1） multicellular spheroid（HCE1/MCS） model and if CD147 mAb can reverse the HCE1/MCS resistance to TAX.Methods HCE1/MCS was obtained by liquid overlay and rotating technique.HCE1/MCS morphological changes were observed before or after the interference of CD147 mAb.The effects of TAX on HCE1/MCS（including inhibition ratio,IC50 and index of multicellular resistance） before or after CD147 mAb treatment were determined by the method of WST-1 and the inhibition ratio curve was mapped.Cell cycle and apoptosis were detected by flow cytometer（FCM）.The expression of CD147 and P-gp of both HCE1/MC and HCE1/MCS was detected by immunocytochemistry.Results HCE1/MCS was established successfully.CD147 mAb could inhibit HCE1/MCS from forming spheroids.CD147 mAb could enhance the sensitivity of HCE1/MCS to TAX.IC50 in different concentrations of CD147 mAb（5,10,20 μg/mL） HCE1/MCS group were（40.31±3.73）,（32.43±1.56）,and（30.69±1.01） μg/mL.CD147 mAb resulted in G1/G0 arrest in HCE1/MCS.CD147 mAb of low concentrations（0-10 μg/mL） caused a dose-dependent inhibition of HCE1/MCS（P0.05）.Combined with TAX,CD147 mAb could also induce HCE1/MCS cell cycle arrest in both G1/S and G2/M stage.The expression of CD147 and P-gp was consistent in HCE1/MCS groups.Conclusion CD147 plays an important role in muliticellular resistance of cervical cancer and inhibition of CD147 can synergistically reverse the multicellular drug resistance（MCR） in cervical cancer.The MCR of HCE1/MCS mediated by CD147 is related to P-gp.

关 键 词：宫颈癌 多细胞球体 CD147 P-糖蛋白 紫杉醇 

分 类 号：R734.2[医药卫生—肿瘤]