摘要
目的诱导对顺铂耐药的人食管鳞癌细胞株,并进一步研究细胞耐药性产生的机制。方法逐渐增加顺铂浓度处理人食管鳞癌细胞HKESC-1,诱导顺铂耐药细胞株HKESC-1/cis。MTT法观察顺铂的细胞毒作用以及细胞的生长曲线。电感耦合等离子体质谱检测细胞内顺铂的蓄积和Pt-DNA加合物的形成。Western blot检测铜离子转运蛋白(copper transporter 1,CTR1)的表达。结果顺铂耐药细胞株HKESC-1/cis较其亲代细胞HKESC-1对顺铂引起的细胞毒作用敏感性下降,耐药指数为2.9。顺铂耐药细胞株HKESC-1/cis与其亲代细胞HKESC-1的生长速度相似。然而,HKESC-1/cis细胞内顺铂蓄积和Pt-DNA加合物的形成较HKESC-1细胞减少,并且CTR1蛋白表达水平降低。结论顺铂通过降低细胞膜CTR1蛋白的表达,抑制顺铂进入细胞,导致细胞耐药性的产生。
Aim To induce cisplatin-resistant esophageal squamous carcinoma cell line and investigate the mechanisms by which the cancer cells develop drug resistance. Methods The cisplatin-resistant subline, designated HKESC-1/eis, was originated by growing parental HKESC-1 cells with gradually increasing doses cisplatin. The cytotoxicity of cisplatin and cell growth curves were determined by MTT assay. Whole-cell cisplatin accumulation and Pt-DNA adduet formation were measured by Inductively Coupled Plasma Mass Spectrometry （ ICP-MS ）. to investigate the protein Western blot was used expression of copper trans-porter 1 （ CTR1 ）. Results HKESC-1/cis was 2. 9 times more resistant to cisplatin-induced cytotoxicity in comparison with its parental cell line HKESC-1. There was no significant difference in growth curves between HKESC-1/cis and HKESC-1. However, compared with HKESC-1 cells, HKESC-1/cis cells exhibited less cisplatin accumulation and Pt-DNA adduct formation, accompanied by decreased CTR1 protein expression. Conclusion Cisplatin induces drug resistant phenotype by decreasing protein level of CTR1, which controls cell accumulation and cytotoxic effect of cisplatin.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2011年第7期911-915,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No81001456)
