Peking University School of Pharmaceutical Sciences, 38 Xueyuan Road, Beijing 100191, Peoples Republic of China.
Cell Biol Int. 2011 Nov;35(11):1159-67. doi: 10.1042/CBI20100870.
This study examined whether Gd (gadolinium) could suppress prostate cancer cell migration and prostate cancer cell-induced osteoclast differentiation. MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide] and colony forming assay showed that GdCl3 treatment inhibited both cell viability and colony forming ability in PC3 cells more significantly than that in DU145 cells. Annexin/PI (propidium iodide) staining showed an increase in apoptotic death of PC3 cells in the presence of GdCl3. Wound healing and adhesion assay indicated that GdCl3 suppressed PC3 cell migration. Western-blot analysis demonstrated that GdCl3 treatment inhibited phosphorylation of ERK (extracellular-signal-regulated kinase) and p38 MAPK (mitogen-activated protein kinase). Pretreatment with PTx (pertussis toxin), a Gi protein inhibitor, conferred resistance to GdCl3-induced colony formation, ERK and p38 phosphorylation in PC3 cells. Moreover, GdCl3 inhibited PC3 cell-induced osteoclast differentiation. RT-PCR (reverse transcription-PCR) indicated that GdCl3 decreased the expression of RANKL (receptor activator of nuclear factor-κB ligand) in PC3 cells, whereas it increased the expression of OPG (osteoprotegerin) in PC3 and DU145 cells. In conclusion, the present study indicated that GdCl3 inhibited PC3 cell migration mediated by the inactivation of both ERK and p38 MAPK pathways via PTx-sensitive G proteins, and also suppressed PC3 cell-induced osteoclast differentiation via regulating the mRNA expression of OPG and RANKL.
本研究旨在探讨钆（Gd）能否抑制前列腺癌细胞迁移和前列腺癌细胞诱导的破骨细胞分化。MTT [3-(4,5-二甲基噻唑-2-基)-2,5-二苯基-2H-四唑溴盐]和集落形成实验表明，GdCl3 处理对 PC3 细胞的细胞活力和集落形成能力的抑制作用明显强于 DU145 细胞。Annexin/PI（碘化丙啶）染色显示 GdCl3 存在时 PC3 细胞凋亡死亡增加。划痕愈合和黏附实验表明 GdCl3 抑制 PC3 细胞迁移。Western-blot 分析表明 GdCl3 抑制 ERK（细胞外信号调节激酶）和 p38 MAPK（丝裂原激活蛋白激酶）的磷酸化。Gi 蛋白抑制剂百日咳毒素（PTx）预处理赋予 PC3 细胞对 GdCl3 诱导的集落形成、ERK 和 p38 磷酸化的抗性。此外，GdCl3 抑制 PC3 细胞诱导的破骨细胞分化。RT-PCR（逆转录-PCR）表明 GdCl3 降低了 PC3 细胞中 RANKL（核因子-κB 配体受体激活剂）的表达，而增加了 PC3 和 DU145 细胞中 OPG（骨保护素）的表达。综上所述，本研究表明 GdCl3 通过 PTx 敏感的 Gi 蛋白抑制 PC3 细胞迁移，通过调节 OPG 和 RANKL 的 mRNA 表达抑制 PC3 细胞诱导的破骨细胞分化。
