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Notch1基因沉默抑制SGC-7901胃癌细胞的增殖和侵袭。

Notch1 silencing inhibits proliferation and invasion in SGC‑7901 gastric cancer cells.

作者信息

Wei Guangbing, Chang Yuanhong, Zheng Jianbao, He Sai, Chen Nanzheng, Wang Xiaolong, Sun Xuejun

机构信息

Department of General Surgery, First Affiliated Hospital of Medical College, Xi'an Jiaotong University, Xi'an, Shaanxi 710061, P.R. China.

Department of Gastroenterology, Xi'an No. 4 Hospital, Affiliated Guangren Hospital, School of Medicine, Xi'an Jiaotong University, Xi'an, Shaanxi 710004, P.R. China.

出版信息

Mol Med Rep. 2014 Apr;9(4):1153-8. doi: 10.3892/mmr.2014.1920. Epub 2014 Jan 28.

Abstract

Downregulation of Notch1 has been shown to exert antineoplastic effects in vivo and in vitro. However, the role of the Notch1 gene in the proliferative and invasive ability of gastric cancer cells is not clear. In this study, we investigated the effect of Notch1 gene silencing on the proliferation and invasion of gastric cancer SGC‑7901 cells. Small interfering RNA (siRNA) targeting Notch1 was transfected into SGC‑7901 cells using Lipofectamine 2000. Proliferation of SGC‑7901 cells was then determined by the MTT assay. Notch1 mRNA expression was determined by reverse transcription‑polymerase chain reaction (RT‑PCR). Invasion of the SGC‑7901 cells was detected by the Transwell assay. The protein levels of cyclin D1, cyclin A1 and cyclin-dependent kinase 2 (CDK2) were determined by western blotting. The mRNA levels of matrix metalloproteinase‑2 (MMP‑2) and cyclooxygenase‑2 (COX‑2) were determined by RT‑PCR. Compared to the control group, the Notch1 mRNA level was significantly decreased following transfection. The growth and invasion rates of SGC‑7901 cells were significantly reduced after Notch1 silencing. Additionally, the expression of cyclin D1 and cyclin A1 proteins and of the MMP‑2 and COX‑2 mRNAs was markedly attenuated. From these results, it was concluded that Notch1 gene silencing inhibits the proliferation of gastric SGC‑7901 cells by decreasing the expression of cyclins D1 and A1, and reduces the invasive ability of SGC‑7901 cells through the downregulation of MMP‑2 and COX‑2 genes. Thus, silencing of the Notch1 pathway may be a novel approach in the treatment of gastrointestinal cancer.

摘要

Notch1的下调已被证明在体内和体外均具有抗肿瘤作用。然而,Notch1基因在胃癌细胞增殖和侵袭能力中的作用尚不清楚。在本研究中,我们调查了Notch1基因沉默对胃癌SGC-7901细胞增殖和侵袭的影响。使用Lipofectamine 2000将靶向Notch1的小干扰RNA(siRNA)转染到SGC-7901细胞中。然后通过MTT法测定SGC-7901细胞的增殖情况。通过逆转录-聚合酶链反应(RT-PCR)测定Notch1 mRNA表达。通过Transwell法检测SGC-7901细胞的侵袭情况。通过蛋白质印迹法测定细胞周期蛋白D1、细胞周期蛋白A1和细胞周期蛋白依赖性激酶2(CDK2)的蛋白质水平。通过RT-PCR测定基质金属蛋白酶-​​2(MMP-2)和环氧合酶-2(COX-2)的mRNA水平。与对照组相比,转染后Notch1 mRNA水平显著降低。Notch1沉默后,SGC-7901细胞的生长和侵袭率显著降低。此外,细胞周期蛋白D1和细胞周期蛋白A1蛋白以及MMP-2和COX-2 mRNA的表达明显减弱。从这些结果得出结论,Notch1基因沉默通过降低细胞周期蛋白D1和A1的表达来抑制胃SGC-7901细胞的增殖,并通过下调MMP-2和COX-2基因来降低SGC-7901细胞的侵袭能力。因此,Notch1通路的沉默可能是治疗胃肠道癌的一种新方法。

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