The Center for Combinatorial Chemistry and Drug Discovery, the School of Pharmaceutical Sciences, Jilin University, Changchun, Jilin, People's Republic of China.
Anticancer Drugs. 2022 Feb 1;33(2):200-207. doi: 10.1097/CAD.0000000000001259.
Drug resistance is a major problem in cancer therapy with cisplatin. It has not been reported that pleiotrophin, which is anti-apoptotic in some cancer cells, is associated with cisplatin resistance. Pleiotrophin was exogenously expressed in 293 cells. Viability and apoptosis of PC3 cells treated with different concentrations of cisplatin in the presence or absence of purified pleiotrophin were determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and flow cytometry, respectively. PC3 cells transfected with shRNAs were analyzed by reverse transcription-polymerase chain reaction (RT-PCR) and western blotting 24 h after transfection. MTT assay data indicated that the EC50 value of cisplatin for PC3 cells was significantly increased in the presence of pleiotrophin. Flow cytometry data demonstrated the pleiotrophin dose-dependent anti-apoptosis in PC3 cells treated with cisplatin. Knockdown of pleiotrophin with sh-RNA, as justified by RT-PCR and western blotting analysis, led to increased cisplatin induced-apoptosis in PC3 cells with an increased level of the cleaved poly ADP-ribose polymerase protein. Pleiotrophin may be a potential antiapoptotic protein associated with cisplatin susceptibility, which warrants further study on the role of pleiotrophin in cisplatin resistance.
顺铂耐药是癌症治疗中的一个主要问题。目前尚未有报道称，某些癌细胞中具有抗细胞凋亡作用的多效蛋白与顺铂耐药有关。我们在 293 细胞中外源性表达了多效蛋白。通过 3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐（MTT）检测和流式细胞术分别检测了不同浓度顺铂处理的 PC3 细胞中存在或不存在纯化的多效蛋白时的细胞活力和细胞凋亡。转染 shRNA 的 PC3 细胞在转染后 24 小时通过逆转录-聚合酶链反应（RT-PCR）和 Western blot 进行分析。MTT 检测数据表明，多效蛋白的存在使 PC3 细胞中顺铂的 EC50 值显著增加。流式细胞术数据表明，多效蛋白剂量依赖性地抑制了顺铂处理的 PC3 细胞的凋亡。通过 RT-PCR 和 Western blot 分析证实，shRNA 敲低多效蛋白导致 PC3 细胞中顺铂诱导的凋亡增加，多聚 ADP-核糖聚合酶蛋白的裂解水平增加。多效蛋白可能是与顺铂敏感性相关的潜在抗细胞凋亡蛋白，这表明有必要进一步研究多效蛋白在顺铂耐药中的作用。
