Li Wentao, Zhai Baoping, Zhi Hui, Li Yuhong, Jia Linjiao, Ding Chao, Zhang Bin, You Wei
Department of Breast Surgery, The People's Hospital of Henan Province (The People's Hospital of Zhengzhou University), Zhengzhou, China, 450000.
Tumour Biol. 2014 Sep;35(9):8883-91. doi: 10.1007/s13277-014-2101-z. Epub 2014 Jun 4.
Docetaxel is a first-line chemotherapeutic agent for treating advanced breast cancer. The development of chemoresistance or multidrug resistance (MDR), however, results in breast cancer chemotherapy failure. This study aims to explore the molecular mechanisms underlying docetaxel-resistance in treatment of breast cancer. The docetaxel-resistant subline MCF7/DOC, derived from the parental sensitive breast cancer cell line MCF7, was established by intermittent exposure to moderate concentrations of docetaxel, followed by examination of its phenotypes. The MCF7/DOC subline showed cross resistance against paclitaxel, doxorubicin, methotrexate, and 5-Fu. Compared to the parental MCF7, MCF7/DOC cells were enlarged with heterogeneous sizes and a cobblestone and polygonal appearance. They were arrested at G2/M phase and proliferated slowly. The colony formation potential of MCF7/DOC in soft agar was significantly increased. MCF7/DOC cells showed reduced intracellular accumulation and increased efflux of rhodamine 123. The mRNA expression level of adenosine triphosphate binding cassette (ABC) transporter family, i.e., ABCB1, ABCC1, ABCC2, ABCG2, and β tubulin isotypes were characterized by quantitative PCR. High-level expression of ABCB1, βI, and βIII tubulin mRNA in MCF7/DOC was detected. Downregulation of ABCB1, βI, and βIII tubulin mediated by three combined siRNAs resulted in stronger growth inhibition of MCF7/DOC than inhibition of the expression of individual genes. ABCB1, βI, and βIII tubulin might contribute to the MDR of MCF7/DOC and be potential therapeutic targets for overcoming MDR of breast cancer.
多西他赛是治疗晚期乳腺癌的一线化疗药物。然而,化疗耐药或多药耐药(MDR)的出现会导致乳腺癌化疗失败。本研究旨在探索乳腺癌治疗中多西他赛耐药的分子机制。通过间歇性暴露于中等浓度的多西他赛,从亲本敏感乳腺癌细胞系MCF7中获得了多西他赛耐药亚系MCF7/DOC,随后对其表型进行了检测。MCF7/DOC亚系对紫杉醇、阿霉素、甲氨蝶呤和5-氟尿嘧啶表现出交叉耐药。与亲本MCF7相比,MCF7/DOC细胞体积增大,大小不均,呈鹅卵石样和多边形外观。它们停滞在G2/M期,增殖缓慢。MCF7/DOC在软琼脂中的集落形成能力显著增强。MCF7/DOC细胞中罗丹明123的细胞内积累减少,流出增加。通过定量PCR对三磷酸腺苷结合盒(ABC)转运蛋白家族即ABCB1、ABCC1、ABCC2、ABCG2和β微管蛋白亚型的mRNA表达水平进行了表征。检测到MCF7/DOC中ABCB1、βI和βIII微管蛋白mRNA的高水平表达。由三种组合的小干扰RNA介导的ABCB1、βI和βIII微管蛋白的下调对MCF7/DOC的生长抑制作用比单个基因表达的抑制作用更强。ABCB1、βI和βIII微管蛋白可能促成了MCF7/DOC的多药耐药,是克服乳腺癌多药耐药的潜在治疗靶点。