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使用小干扰RNA降低结肠癌细胞中烯酰辅酶A水合酶1的表达可抑制细胞增殖和迁移。

Attenuation of enoyl coenzyme A hydratase 1 expression in colorectal cancer cells using small interfering RNA inhibits cell proliferation and migration.

作者信息

Zhao Qing-Mei, Kuang Fei, Wu Han, Zhang Yu-Hao

机构信息

Department of Oncology, Sichuan Mianyang 404 Hospital, Mianyang, Sichuan 621000, P.R. China.

Department of General Surgery, Changhai Hospital of The Second Military Medical University, Shanghai 200433, P.R. China.

出版信息

Mol Med Rep. 2015 Jul;12(1):470-4. doi: 10.3892/mmr.2015.3418. Epub 2015 Mar 4.

Abstract

Colorectal cancer is one of the most commonly diagnosed types of cancer and is a leading cause of cancer-associated mortality worldwide. Short chain enoyl coenzyme A hydratase 1 (ECHS1) is an important gene involved in the mitochondrial fatty acid β-oxidation pathway. In addition, ECHS1 has been implicated in a variety of cancers, including breast, prostate, colon and liver cancer. The aim of the present study was to examine the expression of ECHS1 in the human HCT-8 colorectal cancer cell line. The results showed that ECHS1 expression was significantly increased in poorly-differentiated cells compared with that in well-differentiated cells. In order to further investigate the functions of ECHS1 in colorectal cancer cells, a stably transfected HCT-8 cell line expressing small interfering (si)RNA targeting the ECHS1 gene was established. The expression of the ECHS1 siRNA was found to reduce ECHS1 protein levels in ECHS1-silenced cells by >40%. Cell proliferation and cell migration of the siECHS1 cells were characterized using Cell Counting Kit-8 and Transwell assays, respectively, the results of which showed that the constitutive knockdown of the ECSH1 gene in HCT-8 cells significantly inhibited cell proliferation and migration. Furthermore, decreased levels of Akt and glycogen synthase kinase (GSK)3β phosphorylation were observed in ECHS1-silenced HCT-8 cells compared with that of parental or pU6 empty vector-transfected cells. In conclusion, the results of the present study suggested that ECHS1 may have an important role in colorectal cancer cell proliferation and migration via activation of Akt- and GSK3β-associated signaling pathways.

摘要

结直肠癌是最常被诊断出的癌症类型之一,也是全球癌症相关死亡的主要原因。短链烯酰辅酶A水合酶1(ECHS1)是参与线粒体脂肪酸β氧化途径的一个重要基因。此外,ECHS1与多种癌症有关,包括乳腺癌、前列腺癌、结肠癌和肝癌。本研究的目的是检测ECHS1在人HCT-8结直肠癌细胞系中的表达。结果显示,与高分化细胞相比,低分化细胞中ECHS1的表达显著增加。为了进一步研究ECHS1在结直肠癌细胞中的功能,建立了稳定转染的表达靶向ECHS1基因的小干扰(si)RNA的HCT-8细胞系。发现ECHS1 siRNA的表达使ECHS1沉默细胞中的ECHS1蛋白水平降低了40%以上。分别使用细胞计数试剂盒-8和Transwell实验对siECHS1细胞的细胞增殖和细胞迁移进行了表征,结果表明,HCT-8细胞中ECSH1基因的组成性敲低显著抑制了细胞增殖和迁移。此外,与亲本细胞或转染pU6空载体的细胞相比,在ECHS1沉默的HCT-8细胞中观察到Akt和糖原合酶激酶(GSK)3β磷酸化水平降低。总之,本研究结果表明,ECHS1可能通过激活Akt和GSK3β相关信号通路在结直肠癌细胞增殖和迁移中发挥重要作用。

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