Knockdown of LGR5 suppresses the proliferation of glioma cells in vitro and in vivo.
Knockdown of LGR5 suppresses the proliferation of glioma cells in vitro and in vivo.
复制标题
DOI：
10.3892/or.2013.2826
复制
发表时间：
2014-01
期刊：
影响因子：
4.2
通讯作者：
Zhang Q
中科院分区：
文献类型：
--
作者：
Wang D;Zhou J;Fan C;Jiao F;Liu B;Sun P;Miao J;Zhang Q
富含亮氨酸重复序列的G蛋白偶联受体5（LGR 5）是Wnt信号通路的靶基因之一，最近被鉴定为脑癌干细胞样细胞的标志物。然而，LGR 5在胶质瘤中的作用知之甚少。本研究旨在探讨LGR 5在胶质瘤中的表达与病理分级的关系，以及LGR 5对胶质瘤细胞体内外增殖的影响。首先，我们对54例不同病理分级的胶质瘤进行免疫组化检测，并分析其与Ki-67的相关性。随后，使用蛋白质印迹和qRT-PCR，在三种胶质瘤细胞系U87、U118和U251中评估LGR 5的表达。此外，通过siRNA敲低LGR 5对胶质瘤细胞增殖、细胞周期、克隆形成和体外肿瘤球形成以及体内胶质瘤形成的影响进行了评估。结果显示：① LGR 5在所有胶质瘤标本中均呈阳性表达，其表达随病理分级和Ki-67表达的增高而增高，② LGR 5在3种胶质瘤细胞系中均呈高表达，siRNA可显著降低其表达;和iii）U87细胞中内源性LGR 5的RNAi介导的下调导致细胞增殖的抑制，细胞周期的停滞，并减少体外克隆和肿瘤球形成。此外，LGR 5耗竭显著抑制裸鼠中的肿瘤原位异种移植物生长。这些发现表明LGR 5通过促进肿瘤细胞增殖在胶质瘤形成中起主要作用，表明LGR 5作为病理学的分子标志物和恶性胶质瘤的新治疗靶点。
Leucine-rich repeat containing G protein-coupled receptor 5 (LGR5), one of the target genes of the Wnt signaling pathway, has recently been identified as a marker for brain cancer stem-like cells. However, the role of LGR5 in glioma is poorly understood. The aim of the present study was to investigate the relationship between LGR5 expression and pathological grade in glioma, and the impact of LGR5 on the proliferation of glioma cells in vitro and in vivo. Firstly, LGR5 expression was immunohistochemically evaluated in 54 resected gliomas of different pathologic grades, and its association with Ki-67 was evaluated. Subsequently, using western blotting and qRT-PCR, the expression of LGR5 was assessed in three glioma cell lines U87, U118 and U251. Moreover, the effects of LGR5 knockdown by siRNA on glioma cell proliferation, cell cycle, clone formation and tumorsphere formation in vitro and gliomagenesis in vivo were assessed. The results revealed that i) LGR5 was positively expressed in all glioma specimens and its expression increased with pathologic grade and Ki-67 expression; ii) LGR5 was highly expressed in three glioma cell lines and its expression was reduced significantly by siRNA; and iii) RNAi-mediated downregulation of endogenous LGR5 in U87 cells resulted in the suppression of cell proliferation, arrest of the cell cycle, and reduction in clone and tumorsphere formation in vitro. In addition, LGR5 depletion significantly inhibited tumor orthotopic xenograft growth in nude mice. These findings indicate that LGR5 plays a major role in gliomagenesis by promoting neoplastic cell proliferation, suggesting LGR5 as a molecular marker for pathology and a novel therapeutic target for malignant glioma.
登录
查看更多内容
影响因子：
3.7
作者：
Simon E;Petke D;Böger C;Behrens HM;Warneke V;Ebert M;Röcken C
通讯作者：
Röcken C
影响因子：
4.7
作者：
Scannell CA;Pedersen EA;Mosher JT;Krook MA;Nicholls LA;Wilky BA;Loeb DM;Lawlor ER
通讯作者：
Lawlor ER
影响因子：
8.8
作者：
Barker, Nick;Rookmaaker, Maarten B.;Clevers, Hans
通讯作者：
Clevers, Hans
影响因子：
3.7
作者：
Fukuma, Mariko;Tanese, Keiji;Sakamoto, Michiie
通讯作者：
Sakamoto, Michiie
影响因子：
3.6
作者：
McClanahan, Terrill;Koseoglu, Sandra;Samatar, Ahmed A.
通讯作者：
Samatar, Ahmed A.