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靶向PLK1的短发夹RNA抑制鼻咽癌细胞的增殖和侵袭。

shRNA targeting PLK1 inhibits the proliferation and invasion of nasopharyngeal carcinoma cells.

作者信息

Zhou Yan, Wu Chu, Liu Bingxue, Zhu Juan, Zhong Yating, Yuan Yuqing, Huang Yue, Tang Yunlian

机构信息

Cancer Research Institute, Medical College of Hengyang, Key Laboratory of Tumor Cellular & Molecular Pathology, University of South China, Hengyang, China.

Department of Pathology, People's Hospital of Ningxiang, Changsha, China.

出版信息

Transl Cancer Res. 2020 Sep;9(9):5350-5359. doi: 10.21037/tcr-20-811.

Abstract

BACKGROUND

Polo-like kinase 1 (PLK1) is a serine/threonine protein kinase, which has been studied as a potential gene therapy target for many years. PLK1 is overexpressed in a variety of tumors, and its expression often negatively correlated with patient prognosis. However, the role of PLK1 in nasopharyngeal carcinoma (NPC) is rarely studied.

METHODS

Two recombinant vector plasmids were transfected into CNE2 cell lines by liposome transfection, CNE2/PLK1 shRNA target PLK1 mRNA, as well as a non-targeting control plasmid, CNE2/NC shRNA. Meanwhile, non-transfected cells (CNE2) were also used as controls. Real-time quantitative PCR (qRT-PCR) and Western blotting were performed to detect the transfection effect. The effects of the downregulation of PLK1 on cell biological behavior was evaluated by using CCK8, Transwell, colony-forming and flow-cytometry assays.

RESULTS

PLK1 mRNA and protein were significantly inhibited in CNE2/PLK1 shRNA cells. Compared to control groups, the CNE2/PLK1 shRNA cells showed slower cell growth and a significantly decreased cell-cloning rate. Both migration and invasion were significantly inhibited in experimental cells. The proportions of G2-phase and apoptotic cells within the experimental group were significantly increased.

CONCLUSIONS

Our results indicate that specific interference of gene expression can significantly inhibit the proliferation and invasion of NPC (CNE2) cells.

摘要

背景

Polo样激酶1(PLK1)是一种丝氨酸/苏氨酸蛋白激酶,多年来一直作为潜在的基因治疗靶点进行研究。PLK1在多种肿瘤中过表达,其表达常与患者预后呈负相关。然而,PLK1在鼻咽癌(NPC)中的作用鲜有研究。

方法

通过脂质体转染将两种重组载体质粒转染至CNE2细胞系,CNE2/PLK1 shRNA靶向PLK1 mRNA,以及一种非靶向对照质粒CNE2/NC shRNA。同时,未转染的细胞(CNE2)也用作对照。采用实时定量PCR(qRT-PCR)和蛋白质印迹法检测转染效果。使用CCK8、Transwell、集落形成和流式细胞术检测评估PLK1下调对细胞生物学行为的影响。

结果

CNE2/PLK1 shRNA细胞中PLK1 mRNA和蛋白显著受到抑制。与对照组相比,CNE2/PLK1 shRNA细胞生长较慢,细胞克隆率显著降低。实验组细胞的迁移和侵袭均受到显著抑制。实验组中G2期细胞和凋亡细胞的比例显著增加。

结论

我们的结果表明,基因表达的特异性干扰可显著抑制NPC(CNE2)细胞的增殖和侵袭。

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