Kumazaki Minami, Shinohara Haruka, Taniguchi Kohei, Ueda Hiroshi, Nishi Mayuko, Ryo Akihide, Akao Yukihiro
United Graduate School of Drug Discovery and Medical Information Sciences, Gifu University, Yanagido, Gifu 501-1193, Japan.
Department of Microbiology, Yokohama City University School of Medicine, Yokohama 236-0027, Japan.
Oncotarget. 2015 Sep 22;6(28):25828-42. doi: 10.18632/oncotarget.4558.
Tumor necrosis-factor (TNF)-related apoptosis-inducing ligand (TRAIL) is a member of the TNF-superfamily that selectively induces apoptosis through death receptors (DRs) 4 and/or 5 in cancer cells. These receptors are expressed on the cancer cell surface, without affecting normal cells. Unfortunately, many clinical studies have shown that cancer cells acquire TRAIL-resistance and finally avoid TRAIL-induced apoptosis. The detailed mechanisms of this resistance are not well understood. In the current study, we established a TRAIL-resistant human colon cancer DLD-1 cell line to clarify the mechanisms of TRAIL-resistance and developed agents to cancel its machinery. Also, we found that cancer stem-like cells from breast epithelial proliferating MCF10A cells were also sensitive to TRAIL-induced apoptosis. The enforced expression of DR5 in both TRAIL-resistant cells partially recovered the sensitivity to the TRAIL ligand, which was judged by the activation of caspase-8. As a result, we newly found that the mechanisms of TRAIL-resistance comprised co-existence of a decrease in the expression level of DR5 along with malfunction of its recruitment to the cell surface, as evidenced by Western blot and immunocytological analysis, respectively. Interestingly, α-mangostin, which is a xanthone derivative, canceled the resistance by increasing the expression level of DR5 through down-regulation of miR-133b and effectively induced the translocation of DR5 to the cancer cell surface membrane in TRAIL-resistant DLD-1 cells. These findings indicate that α-mangostin functioned as a sensitizer of TRAIL-induced apoptosis and may thus serve as a possible adjuvant compound for cytokine therapy to conquer TRAIL-resistance.
肿瘤坏死因子(TNF)相关凋亡诱导配体(TRAIL)是TNF超家族的成员,它通过癌细胞中的死亡受体(DRs)4和/或5选择性地诱导细胞凋亡。这些受体在癌细胞表面表达,而不影响正常细胞。不幸的是,许多临床研究表明癌细胞获得了TRAIL抗性,最终避免了TRAIL诱导的细胞凋亡。这种抗性的详细机制尚不清楚。在本研究中,我们建立了一种TRAIL抗性人结肠癌细胞系DLD-1,以阐明TRAIL抗性的机制,并开发消除其机制的药物。此外,我们发现来自乳腺上皮增殖性MCF10A细胞的癌干细胞样细胞对TRAIL诱导的细胞凋亡也敏感。在两种TRAIL抗性细胞中强制表达DR5部分恢复了对TRAIL配体的敏感性,这通过半胱天冬酶-8的激活来判断。结果,我们新发现TRAIL抗性机制包括DR5表达水平降低与其向细胞表面募集功能异常并存,分别通过蛋白质印迹和免疫细胞分析证明。有趣的是,α-山竹黄酮是一种氧杂蒽酮衍生物,它通过下调miR-133b增加DR5的表达水平来消除抗性,并有效地诱导DR5在TRAIL抗性DLD-1细胞中向癌细胞表面膜的转运。这些发现表明α-山竹黄酮作为TRAIL诱导凋亡的敏化剂发挥作用,因此可能作为一种可能的辅助化合物用于细胞因子治疗以克服TRAIL抗性。