Department of Otolaryngology, Huadong Hospital, Fudan University, 221 Yan An Xi Road, Shanghai 200040, China.
Department of Otolaryngology, Huadong Hospital, Fudan University, 221 Yan An Xi Road, Shanghai 200040, China.
Gene. 2014 May 25;542(1):77-82. doi: 10.1016/j.gene.2014.03.020. Epub 2014 Mar 12.
Latent membrane protein 2A (LMP2A) is found to play a key role in the development of nasopharyngeal carcinoma (NPC). However, the role of LMP2A silencing in the inhibition of cell growth of NPC has not been clarified. In this study, we inhibited LMP2A gene expression by lentivirus-mediated RNAi, to explore the effects of LMP2A silencing on the growth of NPC cell line in vitro. A lentivirus-mediated RNAi technology was employed to specifically knock down the LMP2A gene in NPC cell line C666-1. Quantitative real-time polymerase chain reaction, Western blot, flow cytometry and colony formation assays were performed to evaluate the expression of LMP2A and biological behavior of cell line C666-1 in vitro. We successfully construct a highly efficient and stable lentivirus vector, which efficiently downregulate the expression of LMP2A gene in infected cell line C666-1. Down-regulation of the expression of LMP2A significantly inhibits the proliferation and colony formation of C666-1 cells. In addition, the specific down-regulation of LMP2A arrests cells in G0/G1 phase of cell cycle and increases apoptosis rate. Our findings suggest that lentivirus-mediated RNAi knockdown of LMP2A inhibits the growth of NPC cell line C666-1 in vitro, and LMP2A may be a potential target for gene therapy in treatment of NPC.
潜伏膜蛋白 2A（LMP2A）被发现在鼻咽癌（NPC）的发展中起关键作用。然而，LMP2A 沉默在抑制 NPC 细胞生长中的作用尚未阐明。在这项研究中，我们通过慢病毒介导的 RNAi 抑制 LMP2A 基因表达，以探讨 LMP2A 沉默对 NPC 细胞系体外生长的影响。采用慢病毒介导的 RNAi 技术特异性敲低 NPC 细胞系 C666-1 中的 LMP2A 基因。通过定量实时聚合酶链反应、Western blot、流式细胞术和集落形成实验来评估 LMP2A 的表达和细胞系 C666-1 的体外生物学行为。我们成功构建了一种高效稳定的慢病毒载体，可有效下调感染细胞系 C666-1 中 LMP2A 基因的表达。下调 LMP2A 的表达显著抑制了 C666-1 细胞的增殖和集落形成。此外，LMP2A 的特异性下调使细胞周期停滞在 G0/G1 期并增加凋亡率。我们的研究结果表明，慢病毒介导的 RNAi 下调 LMP2A 抑制 NPC 细胞系 C666-1 的体外生长，LMP2A 可能是 NPC 治疗基因治疗的潜在靶点。
