Pu Zhichen, Zhang Xuefeng, Chen Qun, Yuan Xiaolong, Xie Haitang
Department of Clinical Pharmacy, Yijishan Hospital of Wannan Medical College, Anhui Province Center for Drug Clinical Evaluation, Wuhu, Anhui 241001, P.R. China.
Department of Pharmacy, Wuhu Chinese Medicine Hospital, Wuhu, Anhui 241001, P.R. China.
Oncol Rep. 2015 May;33(5):2420-8. doi: 10.3892/or.2015.3864. Epub 2015 Mar 18.
The present study was designed to evaluate the gene polymorphisms of organic anion transporting polypeptide 1B1 (OATP1B1) in predicting the therapeutic efficacy of tamoxifen (TAM) for MCF-7. Established plasmids OATP1Bl wild-type 388GG and 521CC were transfected into MCF-7 cells and used to determine whether the gene polymorphisms affected the therapeutic efficacy of TAM for MCF-7. The established plasmids pcDNA3.1(-)-OATP1B1 wild-type 388GG and 521CC were digested by restriction enzymes and analyzed by gene sequencing. The gene polymorphisms of OATP1Bl in MCF-7 breast cancer cells were examined by RT-PCR and western blot analysis. The results showed that the mutations of OATP1B1 388GG and 521CC led to a decrease of the inhibition and apoptotic rates of MCF-7 cells, albeit not significantly compared to the OATP1B1 group. The G₀/G₁ phase length ratio was reduced, and the S and G₂M phases were increased in the OATP1B1 388GG and 521CC groups, although not significantly compared to the OATP1B1 group. The mutations of OATP1B1 388GG and 521CC inhibited the activity of OATP1B1 protein, restrained the turnover capacity of OATP1B1 and reduced the entrance of TAM into MCF-7 cells, resulting in weakened efficacy of TAM in the treatment of breast cancer.
本研究旨在评估有机阴离子转运多肽1B1（OATP1B1）的基因多态性对预测他莫昔芬（TAM）治疗MCF-7疗效的作用。将已构建的OATP1Bl野生型388GG和521CC质粒转染至MCF-7细胞中，以确定基因多态性是否会影响TAM对MCF-7的治疗效果。对已构建的质粒pcDNA3.1(-)-OATP1B1野生型388GG和521CC进行限制性内切酶消化并通过基因测序进行分析。采用逆转录聚合酶链反应（RT-PCR）和蛋白质免疫印迹分析检测MCF-7乳腺癌细胞中OATP1Bl的基因多态性。结果显示，OATP1B1 388GG和521CC的突变导致MCF-7细胞的抑制率和凋亡率下降，尽管与OATP1B1组相比差异不显著。OATP1B1 388GG和521CC组的G₀/G₁期时长比例降低，S期和G₂M期增加，尽管与OATP1B1组相比差异不显著。OATP1B1 388GG和521CC的突变抑制了OATP1B1蛋白的活性，限制了OATP1B1的转运能力，减少了TAM进入MCF-7细胞，导致TAM治疗乳腺癌的疗效减弱。
