Engür Selin, Dikmen Miriş
a Graduate School of Health Sciences , Anadolu University , Eskisehir , Turkey.
b Faculty of Pharmacy, Department of Pharmacology , Anadolu University , Eskisehir , Turkey.
Acta Clin Belg. 2017 Dec;72(6):391-398. doi: 10.1080/17843286.2017.1302623. Epub 2017 Mar 22.
Proteasome inhibition has recently emerged as a clinically effective anticancer therapeutic approach. The first proteasome inhibitor, bortezomib (Velcade, PS-341), and new proteasome inhibitors including ixazomib have become more important in the development of targeted cancer therapies. Under physiological conditions, MLN9708 (ixazomib citrate), the stable citrate ester drug substance, hydrolyzes rapidly to MLN2238 (ixazomib), the biologically active boronic acid. It is a second-generation proteasome inhibitor, similar to the well-known proteasome inhibitor bortezomib, which is currently being investigated in phase 3 trials as a treatment for multiple Myeloma. Despite the proven efficacy of these drugs in hematologic malignancies, clinical activity is limited to solid tumors such as colon adenocarcinoma. This study is the first to investigate and compare the antiproliferative and apoptotic effects of MLN2238 and bortezomib on human colon adenocarcinoma Caco2 cells. The antiproliferative effects of MLN2238 and bortezomib were determined using WST-1; apoptotic effects of this drug were determined by caspase-3 and a mitochondrial membrane potential (JC-1) activity assay. Expression levels associated with proteasome inhibition and apoptosis of NF-κB and c-myc mRNA were evaluated by RT-PCR. At 24 and 48 h, MLN2238 showed significant time- and concentration-dependent antiproliferative and apoptotic effects on Caco2 cells. Depending on increasing mitochondrial depolarization and caspase-3 activation, MLN2238 induced apoptosis at level similar to that of bortezomib. In addition, MLN2238 downregulated NF-κB and c-myc mRNA expression levels. For the first time, MLN2238 was shown to induce antiproliferative and apoptotic effects on human colon adenocarcinoma cells that are comparable with those of bortezomib; these in vitro data in Caco2 cells support the development of MLN2238 for colon cancer.
蛋白酶体抑制作用最近已成为一种临床有效的抗癌治疗方法。首个蛋白酶体抑制剂硼替佐米（万珂，PS - 341）以及包括伊沙佐米在内的新型蛋白酶体抑制剂在靶向癌症治疗的发展中变得愈发重要。在生理条件下，稳定的柠檬酸酯原料药MLN9708（枸橼酸伊沙佐米）会迅速水解为具有生物活性的硼酸MLN2238（伊沙佐米）。它是一种第二代蛋白酶体抑制剂，与著名的蛋白酶体抑制剂硼替佐米类似，目前正处于3期试验阶段，用于治疗多发性骨髓瘤。尽管这些药物在血液系统恶性肿瘤中已证实具有疗效，但其临床活性在实体瘤如结肠腺癌中仍有限。本研究首次探究并比较了MLN2238和硼替佐米对人结肠腺癌Caco2细胞的抗增殖和凋亡作用。使用WST - 1测定MLN2238和硼替佐米的抗增殖作用；通过半胱天冬酶 - 3和线粒体膜电位（JC - 1）活性测定来确定该药物的凋亡作用。通过逆转录聚合酶链反应（RT - PCR）评估与蛋白酶体抑制以及核因子κB（NF - κB）和原癌基因c - myc mRNA凋亡相关的表达水平。在24小时和48小时时，MLN2238对Caco2细胞显示出显著的时间和浓度依赖性抗增殖和凋亡作用。随着线粒体去极化增加和半胱天冬酶 - 3激活，MLN2238诱导凋亡的水平与硼替佐米相似。此外，MLN2238下调了NF - κB和c - myc mRNA表达水平。首次证明MLN2238对人结肠腺癌细胞具有与硼替佐米相当的抗增殖和凋亡作用；这些在Caco2细胞中的体外数据支持了MLN2238用于结肠癌的研发。
