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

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

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摘要

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 apop-totic 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.
机译:在本研究中,使用电穿孔沉默EC109人食道癌细胞系中ATP结合盒蛋白E1(ABCE1)的基因表达,以检查ABCE1基因是否对癌细胞的生长迁移和细胞周期产生影响。设计并合成了ABCE1的小干扰(si)RNA序列,以通过电穿孔转染EC109细胞。然后通过逆转录定量聚合酶链反应(RT-qPCR)和蛋白质印迹分析检测ABCE1的mRNA和蛋白质表达水平。使用流式细胞仪进行细胞周期和凋亡的分析。使用细胞计数试剂盒8(CCK-8)以及增殖,伤口愈合和细胞侵袭试验评估了ABCE1基因沉默对EC109人食道癌细胞增殖,迁移和侵袭能力的影响。实验组ABCE1的mRNA和蛋白表达水平明显低于对照组(P <0.05)。实验组细胞凋亡率明显高于对照组和空白组(P <0.01)。 CCK-8增殖实验表明,与对照组和空白组相比,实验组EC109细胞的增殖受到明显抑制(P <0.05)。伤口愈合试验表明,实验组细胞的迁移能力明显降低(P <0.05)。 Transwell试验箱检测结果表明,实验组EC109细胞的侵袭能力明显降低(P <0.01)。这些结果表明,ABCE1与食管癌中的细胞增殖,侵袭和迁移密切相关,通过电穿孔使ABCE1基因沉默可以显着降低体外EC109细胞的增殖,侵袭和迁移能力。

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