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RNAi 介导的 SYT14 敲低抑制人神经胶质瘤细胞系 U87MG 的生长。

RNAi-mediated SYT14 knockdown inhibits the growth of human glioma cell line U87MG.

机构信息

Department of Neurosurgery, The First Affiliated Hospital of Wannan Medical College, Wuhu City, Anhui, 241001, China.

Department of Physiology, School of Basic Medicine, Wannan Medical College, Wuhu City, Anhui, 241000, China.

出版信息

Brain Res Bull. 2018 Jun;140:60-64. doi: 10.1016/j.brainresbull.2018.04.002. Epub 2018 Apr 7.

Abstract

SYT14 (Synaptotagmin 14) participates in pathomechanical neurodegeneration and contributes to abnormal neurodevelopment. However, the functional mechanism of SYT14 in human glioma tumorigenesis remains unclear. In the present study, we measured the expression levels of SYT14 mRNA in human glioma cell lines, U373MG, U178, and U87MG and neural stem cells (NSC) cell line by RT-PCR, and used lentivirus-mediated small hairpin RNAs (shRNAs) to knock down SYT14 expression in U87MG cells. Changes in SYT14 expression were determined by real-time PCR. Cell proliferation and colony formation assays were used to analyze the role of SYT14 in U87MG cell proliferation, and cell apoptosis was assessed by flow cytometry. SYT14 mRNA expression was detected in the three glioma cell lines, and was highest in the U87MG cell line. The RNAi-mediated knockdown of SYT14 significantly decreased cell proliferation and colony formation in U87MG cells, and caused a moderate increase in apoptosis. Fewer S phase cells and more G2/M phase cells were observed. These data indicate that SYT14 is highly expressed in glioma cells, and may participate in glioma cell proliferation, apoptosis, and colony formation.

摘要

SYT14(突触结合蛋白 14)参与病理性神经退行性变,并有助于异常神经发育。然而,SYT14 在人类神经胶质瘤肿瘤发生中的功能机制尚不清楚。在本研究中,我们通过 RT-PCR 测量了 SYT14 mRNA 在人神经胶质瘤细胞系 U373MG、U178 和 U87MG 以及神经干细胞(NSC)细胞系中的表达水平,并使用慢病毒介导的短发夹 RNA(shRNA)敲低 U87MG 细胞中的 SYT14 表达。通过实时 PCR 确定 SYT14 表达的变化。细胞增殖和集落形成实验用于分析 SYT14 在 U87MG 细胞增殖中的作用,通过流式细胞术评估细胞凋亡。在三种神经胶质瘤细胞系中检测到 SYT14 mRNA 表达,在 U87MG 细胞系中表达最高。SYT14 的 RNAi 介导的敲低显着降低了 U87MG 细胞的增殖和集落形成,并导致适度增加的细胞凋亡。观察到更少的 S 期细胞和更多的 G2/M 期细胞。这些数据表明,SYT14 在神经胶质瘤细胞中高表达,可能参与神经胶质瘤细胞的增殖、凋亡和集落形成。

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