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通过电穿孔沉默ATP结合盒蛋白E1基因对EC109人食管癌细胞增殖和迁移的影响

Effects of silencing the ATP-binding cassette protein E1 gene by electroporation on the proliferation and migration of EC109 human esophageal cancer cells.

作者信息

Li Xiao-Rui, Yang Liu-Zhong, Huo Xiao-Qing, Wang Ying, Yang Qing-Hui, Zhang Qing-Qin

机构信息

Department of Oncology, The First Affiliated Hospital of Xinxiang Medical College, Weihui, Henan 453100, P.R. China.

出版信息

Mol Med Rep. 2015 Jul;12(1):837-42. doi: 10.3892/mmr.2015.3512. Epub 2015 Mar 19.

Abstract

In the present study, the gene expression of ATP-binding cassette protein E1 (ABCE1) in the EC109 human esophageal cancer cell line was silenced using electroporation to examine the effect if the ABCE1 gene on the growth migration and cell cycle of cancer cells. The small interference (si)RNA sequence of ABCE1 was designed and synthesized to transfect the EC109 cells by electroporation. The mRNA and protein expression levels of ABCE1 were then detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot analysis. The analysis of the cell cycle and apoptosis was performed using flow cytometry. The effect of silencing the ABCE1 gene on the proliferation, migration and invasive ability of the EC109 human esophageal cancer cells were assessed using a Cell counting kit-8 (CCK-8) and with proliferation, wound-healing and cell invasion assays. The mRNA and protein expression levels of ABCE1 were significantly lower in the experimental group compared with the control group (P<0.05). The apoptotic rate of the experimental group was markedly higher than the control group and blank group (P<0.01). The CCK-8 proliferation assay revealed that, compared with the control and blank groups, the proliferation of the EC109 cells in the experimental group was significantly inhibited (P<0.05). The wound healing assay revealed that the migration capacity of the cells in the experimental group was significantly decreased (P<0.05). The Transwell chamber assay demonstrated that the invasive ability of the EC109 cells in the experimental group was significantly decreased (P<0.01). These results revealed that ABCE1 is closely associated with cell proliferation, invasion and migration in esophageal cancer and silencing the ABCE1 gene by electroporation can significantly reduce the proliferation, invasion and migration capacity of EC109 cells in vitro.

摘要

在本研究中,采用电穿孔法使人类食管癌EC109细胞系中的ATP结合盒蛋白E1(ABCE1)基因表达沉默,以研究ABCE1基因对癌细胞生长、迁移及细胞周期的影响。设计并合成ABCE1的小干扰(si)RNA序列,通过电穿孔法转染EC109细胞。然后采用逆转录定量聚合酶链反应(RT-qPCR)和蛋白质免疫印迹分析检测ABCE1的mRNA和蛋白表达水平。使用流式细胞术进行细胞周期和凋亡分析。采用细胞计数试剂盒-8(CCK-8)以及增殖、伤口愈合和细胞侵袭实验,评估沉默ABCE1基因对人食管癌EC109细胞增殖、迁移和侵袭能力的影响。与对照组相比,实验组中ABCE1的mRNA和蛋白表达水平显著降低(P<0.05)。实验组的凋亡率明显高于对照组和空白组(P<0.01)。CCK-8增殖实验显示,与对照组和空白组相比,实验组中EC109细胞的增殖受到显著抑制(P<0.05)。伤口愈合实验表明,实验组细胞的迁移能力显著下降(P<0.05)。Transwell小室实验表明,实验组中EC109细胞的侵袭能力显著降低(P<0.01)。这些结果表明,ABCE1与食管癌细胞的增殖、侵袭和迁移密切相关,通过电穿孔法沉默ABCE1基因可显著降低EC109细胞在体外培养时的增殖、侵袭和迁移能力。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/9879/4438962/44b5e675ee42/MMR-12-01-0837-g00.jpg

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