人乳腺癌MCF-7他莫昔芬耐药细胞株的建立及鉴定.docVIP

人乳腺癌MCF-7他莫昔芬耐药细胞株的建立及鉴定 莫志强，涂刚，杨光伦 （400016 重庆医科大学附属第一医院） [摘要] 目的 构建人乳腺癌细胞MCF-7他莫昔芬（Tamoxifen,TAM）耐药细胞模型，并鉴定其耐药性。方法 采用高浓度短时间4-羟他莫昔芬(4-hydroxytamoxifen，OHT)冲击法，诱导人乳腺癌MCF-7/TAM耐药细胞株。通过细胞形态学观察、生长曲线、细胞倍增时间、药物毒性试验、细胞周期及凋亡相关蛋白BIK和Caspase-7的检测，评价MCF-7/TAM耐药细胞株的耐药性。结果 成功构建MCF-7/TAM耐药细胞株，其耐药指数(resistance index，RI)为13.88±2.93 (P0.05)。与其母细胞相比，MCF-7/TAM耐药细胞株不仅细胞形态发生了变化，生长增殖能力增强(P0.05)，细胞倍增时间缩短(P0.05)，而且对OHT介导的毒性效应有较强的抵抗能力(P0.05)，OHT不能促进MCF-7/TAM耐药细胞内BIK和Caspase-7蛋白的表达(P0.05)。结论 通过高浓度短时间OHT冲击法诱导的MCF-7/TAM耐药细胞株对TAM产生一定的耐药性。 [关键] 乳腺癌； 他莫昔芬； 耐药细胞株 [中图分类号] R737.9 [文献标志码] A Establishment and identificantion of Tamoxifen-resitant subline of MCF-7 human breast cancer cell lineMo Zhiqiang, Tu Gang, Luo haojun , Li zhenhua, Yang Guanglun (Department of Endocrine breast surgery, the 1st Affiliated Hospital, Chongqing Medical University, Chongqing 400016, China) [Abstract] Objective To establish Tamoxifen (TAM) -resitant subline of MCF-7 human breast cancer cell and investigate its drug tolerance. Methods MCF-7 Tamoxifen-resistant(MCF-7/TAM-R) cell line was derived by clonal selection in high concentration of 4-hydroxytamoxifen(OHT) for a short period . Drug tolerance of MCF-7/TAM-R cells were characterized by investigating cells morphology, cellular growth curve, population doubling time, drug toxicity tests, cell cycle and apoptosis-related protein of BIK and Caspase-7 expression. Results MCF-7/TAM-R cell line had been established, and its resistance index (RI) was up to 13.88±2.93(P0.05). Compared to its parental cell line, MCF-7/TAM-R cell line was varied in morphology, increased in proliferation (P0.05), decreased in population doubling time(P0.05) and showed no respondence to the cytotoxic effects of OHT (P0.05). In MCF-7/TAM-R cell line, OHT failed to induce the BIK and Caspase-7 expression(P0.05). Conclusion MCF-7/TAM-R cell line derived by clonal selection in high concentration of OHT for a short period showed a significant resisitance to TAM. [Key words] br