School of Medicine, Nankai University, Tianjin, China.
Eur Rev Med Pharmacol Sci. 2019 Apr;23(7):2880-2887. doi: 10.26355/eurrev_201904_17566.
To detect the expression of long non-coding ribonucleic acid (lncRNA) plasmacytoma variant translocation gene 1 (PVT1) in uveal melanoma (UM) tissues, and to investigate its influence on the proliferation and apoptosis of UM cells as well as its mechanism.
40 cases of UM tissues and 40 cases of adjacent tissues surgically resected in our hospital from October 2015 to April 2018 were collected. The expression level of lncRNA PVT1 in these tissues was determined by Reverse Transcription-Polymerase Chain Reaction (RT-PCR). Stable knockdown of lncRNA PVT1 was constructed in human UM cell line OCM-1 using small interfering RNA (siRNA). The impact of lncRNA PVT1 on UM cell proliferation was detected by Cell Counting kit-8 (CCK-8) and colony formation assay. Flow cytometry was applied to measure the apoptotic level of UM cells in the blank control group and lncRNA PVT1 knockdown group. Meanwhile, the expression level of enhancer of zeste homologue 2 (EZH2) was determined by Western blotting.
The expression level of lncRNA PVT1 in UM tissues was remarkably higher than that in the adjacent tissues (p<0.05). UM cell proliferation was notably repressed after lncRNA PVT1 knockdown by siRNA. Flow cytometry results indicated that the number of apoptotic UM cells in lncRNA PVT1 knockdown group significantly increased compared with that in the blank control group (p<0.05). The protein expression of EZH2 was suppressed after lncRNA PVT1 knockdown (p<0.05).
LncRNA PVT1 knockdown in UM cells can repress the proliferation of UM cells and promote their apoptosis by regulating EZH2 expression.
检测长链非编码核糖核酸(lncRNA)浆细胞瘤变异易位基因 1(PVT1)在葡萄膜黑色素瘤(UM)组织中的表达,探讨其对UM 细胞增殖和凋亡的影响及其机制。
收集我院 2015 年 10 月至 2018 年 4 月手术切除的 40 例 UM 组织和 40 例相邻组织。采用逆转录-聚合酶链反应(RT-PCR)检测这些组织中 lncRNA PVT1 的表达水平。采用小干扰 RNA(siRNA)构建人 UM 细胞系 OCM-1 中 lncRNA PVT1 的稳定敲低。CCK-8 和集落形成实验检测 lncRNA PVT1 对 UM 细胞增殖的影响。流式细胞术检测空白对照组和 lncRNA PVT1 敲低组 UM 细胞的凋亡水平。同时,采用 Western blot 检测增强子的表达水平 2 同源物(EZH2)。
UM 组织中 lncRNA PVT1 的表达水平明显高于相邻组织(p<0.05)。siRNA 敲低 lncRNA PVT1 后,UM 细胞增殖明显受到抑制。流式细胞术结果表明,lncRNA PVT1 敲低组 UM 细胞凋亡数明显高于空白对照组(p<0.05)。lncRNA PVT1 敲低后 EZH2 蛋白表达受到抑制(p<0.05)。
lncRNA PVT1 敲低可通过调节 EZH2 表达抑制 UM 细胞增殖,促进其凋亡。