Lentiviral-mediated overexpression of KCTD12 inhibits the proliferation of human uveal melanoma OCM-1 cells
Lentiviral-mediated overexpression of KCTD12 inhibits the proliferation of human uveal melanoma OCM-1 cells
复制标题
慢病毒介导的KCTD12过表达抑制人葡萄膜黑色素瘤OCM-1细胞的增殖
DOI：
10.3892/or.2016.5325
复制
发表时间：
2017-02-01
期刊：
影响因子：
4.2
通讯作者：
Jin, Jingji
中科院分区：
文献类型：
--
作者：
Luo, Lifu;Cui, Jizhe;Jin, Jingji
人钾通道四聚化结构域12（KCTD 12，也称为Pfetin）是KCTD家族的一员，该家族由26个成员组成。据报道，KCTD 12调节GABA（B）受体信号传导的激动剂效力和动力学。蛋白质组学分析表明KCTD 12可能是胃肠道间质瘤诊断和预后的潜在生物标志物。然而，关于KCTD 12在其他肿瘤类型中的作用的报道很少。在本研究中，我们设计并亚克隆到pLVX-Puro载体的N-末端Flag标记的人KCTD 12。然后，我们产生了稳定表达KCTD 12的人葡萄膜黑色素瘤细胞系（OCM-1）。利用此稳定的细胞系，我们进行了一系列实验，包括集落形成，侵袭，迁移和伤口愈合测定，流式细胞术和蛋白质印迹。基于实验结果，我们首次证明KCTD 12有效地抑制了OCM-1细胞的增殖，并限制了OCM-1细胞的扩散。在流式细胞仪分析中，观察到KCTD 12过表达的OCM-1细胞中G2/M向G1期进展的延长。此外，在裸鼠中观察到KCTD 12过表达的OCM-1细胞异种移植物生长的抑制。总之，KCTD 12可能作为葡萄膜黑色素瘤患者的新的治疗靶点。
Human potassium channel tetramerization domain containing 12 (KCTD12, also known as Pfetin) is a member of the KCTD family which consists of 26 members. It has been reported that KCTD12 regulates agonist potency and kinetics of GABA(B) receptor signaling. Proteomic analysis indicates that KCTD12 may be a potential biomarker for the diagnosis and prognosis of gastrointestinal stromal tumors. However, little has been reported concerning the role of KCTD12 in the other tumor types. In the present study, we designed and subcloned N-terminally Flag-tagged human KCTD12 into the pLVX-Puro vector. We then generated a human uveal melanoma cell line (OCM-1) stably expressing KCTD12. Using this stable cell line, we performed a series of experiments including colony formation, invasion, migration and wound healing assays, flow cytometry and western blotting. Based on the experimental results, we first demonstrated that KCTD12 effectively suppressed the proliferation of OCM-1 cells and limited the spread of OCM-1 cells. In the flow cytometric analysis, prolongation of the progression of G2/M to G1 phase in the KCTD12-overexpressing OCM-1 cells was observed. In addition, inhibition of KCTD12-overexpressing OCM-1 cell xenograft growth in nude mice was observed. Taken together, KCTD12 may serve as a novel therapeutic target for patients with uveal melanoma.