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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 a human breast cancer drug-resistant cell line MCF-7 / ADM and analyze its biological characteristics preliminarily. Methods Adriamycin (ADM) -induced drug and human breast cancer cell line MCF-7 were used to induce MCF-7 / ADM cells by using low concentration continuous induction method and compared with MTT method Cell growth curve, population doubling time change, clone formation assay to detect cell clone formation ability, and immunocytochemical staining to analyze the expression of cell biochemical markers. Results The resistant cell line MCF-7 / ADM had a doxorubicin resistance index of 9.1. After 60 days of withdrawal, the drug resistance index remained above 7.0 and its resistance was stable. The drug-resistant cell line was doubled in withdrawal culture time Compared with the parental cells of MCF-7, the number of colonies and the volume of colonies were significantly different (P <0.01). Immunocytochemical staining showed that the positive expression of HER-2 and ALDH1 protein in the resistant cells was significant Higher than the parental cells MCF-7 (P <0.05), ER protein expression was significantly lost than the parental cells (P <0.05). CONCLUSION: The established MCF-7 / ADM model has the basic biological characteristics of drug-resistant cells. The ability of independent growth and self-proliferation in vitro is obviously enhanced. The dedifferentiation tendency is obvious. The effectiveness of endocrine therapy is greatly reduced, and the malignancy is significantly increased.