西妥昔单抗体外增强人鼻咽癌细胞对紫杉醇化疗敏感性的实验研究  

Experimental study of Cetuximab in improving chemotherapy sensitivity of paclitaxel on human nasopharyngeal carcinoma cell line in vitro.

作　　者：陈新 刘丹 CHEN Xin;LIU Dan(Department of Otolaryngology,Hubei Provincial Hospital of Integrated Traditional Chinese&Western Medicine,Wuhan Hubei 430015,China.)

机构地区：[1]湖北省中西医结合医院耳鼻喉科,湖北武汉430015

出　　处：《临床和实验医学杂志》2020年第12期1287-1291,共5页Journal of Clinical and Experimental Medicine

基　　金：湖北省教学研究基金资助项目(编号:2016231)。

摘　　要：目的观察西妥昔单抗(C225)体外增强人鼻咽癌CNE-1细胞对紫杉醇(PTX)化疗敏感性作用,并探讨可能的作用机制。方法将人鼻咽癌CNE-1细胞株随机分为对照组、PTX组、联合用药组,PTX组采用30μg/ml PTX处理,联合用药组采用30μg/ml PTX+250μg/ml C225处理,对照组加入等体积的二甲基亚砜(DMSO)。MTT法检测细胞的PTX耐药性,取对数期的CNE-1细胞分为9组,4组分别加入浓度为0.01μmol/L、0.1μmol/L、1μmol/L、10μmol/L的PTX;另有4组也加入PTX,同时加入C225,使C225最终浓度为250μg/ml,空白对照组中加入等体积的DMSO,培养48 h。MTT法检测细胞抑制率,流式细胞仪检测细胞凋亡率及细胞周期分布,Western blotting、RT-PCR分别检测细胞周期蛋白D1(CyclinD1)、B淋巴细胞瘤-2(Bcl-2)、BCL2-相关X蛋白(Bax)蛋白水平及其基因相对表达量。结果PTX组CNE-1细胞抑制率为(15.99±2.13)%,联合用药组CNE-1细胞抑制率为37.10%(37.10±2.48)%,差异有统计学意义(t=11.184,P=0.000)。与对照组相比,PTX组、联合用药组的细胞凋亡率、G0~G1期细胞比率均升高(P<0.05),而S期细胞比率降低(P<0.05)。与PTX相比,联合用药组的细胞凋亡率、G0~G1期细胞比率升高(P<0.05),S期细胞比率与M期细胞比率均明显降低(P<0.05)。MTT检测PTX耐药性结果显示,空白对照组、C225组细胞的细胞活性随着PTX浓度增加而降低,空白对照组、C225组CNE-1细胞的PTX IC 50分别为4.179μmol/L、0.25μmol/L。与对照组相比,PTX组、联合用药组CyclinD1、Bcl-2蛋白水平与基因相对表达量均明显降低(P<0.05),而Bax蛋白水平与基因相对表达量均明显升高(P<0.05)。与PTX相比,联合用药组CyclinD1、Bcl-2蛋白水平与基因相对表达量均明显降低(P<0.05),而Bax蛋白水平与基因相对表达量均明显升高(P<0.05)。结论在PTX化疗的基础上增加C225处理,可以促进PTX对人鼻咽癌CNE-1细胞的增殖抑制作用以及促凋亡作用,降低细胞的CNE-1细胞的PTX IC 50Objective To observe the effects of Cetuximab(C225)in improving chemotherapy sensitivity of Paclitaxel(PTX)on human nasopharyngeal carcinoma CNE-1 cell line in vitro,and to explore the possible mechanism.Methods Human nasopharyngeal carcinoma CNE-1 cell lines were randomly divided into control group,PTX group and drug combination group.PTX group was treated with 30μg/ml PTX,and drug combination group was treated with 30μg/mL PTX+250μg/mL C225,and control group was added with equal volume of DMSO.The PTX resistance of the cells was detected by MTT assay.The logarithmic CNE-1 cells were divided into 9 groups,and 4 groups were added with concentrations of PTX 0.01μmol/L,0.1μmol/L,1μmol/L,10μmol/L;Another 4 groups also added PTX and C225 at the same time,so that the final concentration of C225 was 250μg/ml,and an equal volume of DMSO was added to the blank control group,and cultured for 48 h.The cell inhibition rate were detected by MTT assay.The apoptosis rate and cell cycle distribution were detected by flow cytometry.Western blotting and RT-PCR were used to detect the protein levels and gene relative expression levels of CyclinD1,B lymphocytoma-2(Bcl-2)and BCL2-related X protein(Bax).Results The inhibition rate of CNE-1 cells was(15.99±2.13)%in PTX group and was(37.10±2.48)%in drug combination group(t=11.184,P=0.000).Compared with control group,the apoptosis rate and the proportion of cells in G0-G1 phase in PTX group and drug combination group were all increased(P<0.05)while the proportion of cells in S phase was decreased(P<0.05).Compared with PTX group,the apoptosis rate and the proportion of cells in G0-G1 phase in drug combination group were increased(P<0.05)while the proportions of cells in S phase and cells in M phase were decreased significantly(P<0.05).MTT results of detecting PTX resistance showed that the cell activity in control group and C225 group were decreased with the increase of PTX concentration,and the PTX IC 50 of CNE-1 cells was 4.179μmol/L in control group and 0.25μmol/L in C225

关 键 词：人鼻咽癌CNE-1细胞 西妥昔单抗 紫杉醇 细胞增殖 细胞凋亡 

分 类 号：R739.63[医药卫生—肿瘤]