沉默尿路上皮癌相关蛋白 1 抑制髓母细胞瘤细胞的增殖和迁移。

Silencing of Urothelial Carcinoma Associated 1 Inhibits the Proliferation and Migration of Medulloblastoma Cells.

机构信息

Department of Neurosurgery, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong, China (mainland).

出版信息

Med Sci Monit. 2017 Sep 16;23:4454-4461. doi: 10.12659/msm.904675.

Abstract

BACKGROUND UCA1 is a long non-coding RNA that has been found to be aberrantly upregulated in various cancers. The aim of this study was to determine the expression level and function of UCA1 in medulloblastoma, the most common malignant brain tumor during childhood. MATERIAL AND METHODS Real-time PCR was used to detect the expression of UCA1 in medulloblastoma specimens and cell lines. Lentiviral-mediated expression of a short hairpin RNA (shRNA) targeting UCA1 or a negative control shRNA was also achieved with the medulloblastoma cell line, Daoy. Cell proliferation and cell cycle progression were subsequently characterized with cell counting kit (CCK)-8 and flow cytometry. Cell migration was examined in wound healing and Transwell migration assays. RESULTS Levels of UCA1 mRNA were higher in the medulloblastoma specimens (p<0.05) and cell lines (p<0.05) compared to the corresponding nontumor adjacent tissue specimens and a glioblastoma cell line, respectively. For the Daoy cells with silenced UCA1, their proliferation was reduced by 30% compared to the Daoy cells expressing a negative control shRNA (p=0.017). Cell cycle arrest in the G0/G1 phase, resulting in a decreased number of cells in the S phase, as well as reduced cell migration in both wound scratch healing (p=0.001) and Transwell migration assays (p=0.021) were also observed for the Daoy cells with silenced UCA1. CONCLUSIONS UCA1 was highly expressed in part of medulloblastoma specimens and cell lines examined. In addition, knockdown of UCA1 significantly inhibited the proliferation and migration of medulloblastoma cells in vitro.

摘要

背景 UCA1 是一种长链非编码 RNA,已在各种癌症中发现异常上调。本研究旨在确定 UCA1 在儿童中最常见的恶性脑瘤——髓母细胞瘤中的表达水平和功能。

材料与方法 采用实时 PCR 检测髓母细胞瘤标本和细胞系中 UCA1 的表达。通过慢病毒介导表达针对 UCA1 的短发夹 RNA(shRNA)或阴性对照 shRNA,实现对髓母细胞瘤细胞系 Daoy 的靶向沉默。随后,通过细胞计数试剂盒(CCK)-8 和流式细胞术检测细胞增殖和细胞周期进程。通过划痕愈合和 Transwell 迁移实验检测细胞迁移。

结果 与相应的非肿瘤邻近组织标本和胶质母细胞瘤细胞系相比,髓母细胞瘤标本(p<0.05)和细胞系(p<0.05)中 UCA1 mRNA 水平更高。与表达阴性对照 shRNA 的 Daoy 细胞相比,沉默 UCA1 的 Daoy 细胞的增殖减少了 30%(p=0.017)。沉默 UCA1 的 Daoy 细胞的细胞周期阻滞在 G0/G1 期,导致 S 期细胞数量减少,以及划痕愈合(p=0.001)和 Transwell 迁移实验(p=0.021)中的细胞迁移减少。

结论 UCA1 在部分髓母细胞瘤标本和细胞系中高表达。此外,沉默 UCA1 可显著抑制髓母细胞瘤细胞的体外增殖和迁移。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cfbf/5612201/5db425070396/medscimonit-23-4454-g001.jpg

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