摘要
目的建立人乳腺癌耐药细胞系MCF-7/ADM,对其生物学特性进行初步分析评价。方法以阿霉素(ADM)为诱导药物,人乳腺癌细胞系MCF-7为诱导对象,采用低浓度持续加量诱导方法,建立人乳腺癌耐药细胞系MCF-7/ADM,MTT法比较细胞生长曲线、群体倍增时间改变,克隆形成实验检测细胞克隆形成能力,免疫细胞化学染色分析细胞生化标志物的表达情况。结果建成的MCF-7/ADM耐药细胞株,耐阿霉素指数为9.1,撤药培养60 d后耐药指数仍维持在7.0以上,耐药性稳定,耐药细胞株撤药培养倍增时间较亲本细胞明显缩短,平板集落数目、集落的体积与MCF-7亲代敏感细胞比较差异明显(P<0.01);细胞免疫化学染色分析显示耐药细胞中,HER-2、ALDH1蛋白的阳性表达明显高于亲代细胞MCF-7(P<0.05),ER蛋白的表达较亲代细胞明显丢失(P<0.05)。结论建立的MCF-7/ADM模型具有耐药细胞的基本生物学特性,体外独立生长和自我增殖的能力明显增强,去分化趋势明显,对内分泌治疗的有效性大大降低,恶性程度显著增高。
Objective To establish breast cancer drug resistance cell line MCF -7/ADM and analyze its biologic features. Methods A new breast cancer drug resistant cell line MCF -7/ADM was established by stepwise increasing concentration of adriamycin. Ceil growth curve and doubling time change were analyzed by MTY method. Colony - forming capacity was tested by colony formation assay, and immnno - cytochemistry staining was adopted to analyze the biological characteristics of the new breast cancer drug resistant cell line MCF - 7/ADM. Results The breast cancer adriamycin - resistant cell line MCF - 7/ADM was successfully induced. The resistance index was 9.1. Comparing with the parental ADM - sensitive cells, the resistance in- dex of multi - drug resistant cell line MCF - 7/ADM was kept above 7.0, showing stable drug resistance, shorter withdrawal culture doubling time, and significant difference in plate colony count, volume of colony（P 〈0.01 ）. Immunocytochemical stai- ning analysis showed that the positive expression rates of HER - 2 and ALDH1 protein were higher in multi - drug resistant cell line MCF -7/ADM than those in parental MCF -7 cells （P 〈 0.05 ）, and ER protein expression was obviously lost compared to parental MCF -7 cells. Conclusion The cell line MCF -7/ADM established in this study has basic biological characteristics of muhidrug - resistant cells, indicating increasing independent growth capacity and self - proliferation capacity in vitro, obvious dedifferentiation trend, poor curative effect on endocrinotherapy, and the worsening condition of breast cancer.
出处
《中国卫生检验杂志》
北大核心
2014年第4期499-502,共4页
Chinese Journal of Health Laboratory Technology
基金
杭州市科技局基金资助(20100633B16)
关键词
乳腺癌
MCF-7
ADM
耐药
生物学特性
Breast cancer
MCF-7/ADM
Drug resistance
Biological characteristics
