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Skp2的干扰有效抑制结肠癌的发展和转移。

Interference of Skp2 effectively inhibits the development and metastasis of colon carcinoma.

作者信息

Chen Haijin, Mo Xiaodong, Yu Jinlong, Huang Shuxin, Huang Zonghai, Gao Liping

机构信息

Department of General Surgery, Zhujiang Hospital, Southern Medical University, Guangzhou, Guangdong 510280, P.R. China.

Department of Gastrointestinal Surgery, PLA No. 101 Hospital, Wuxi, Jiangsu 214044, P.R. China.

出版信息

Mol Med Rep. 2014 Aug;10(2):1129-35. doi: 10.3892/mmr.2014.2308. Epub 2014 Jun 10.

Abstract

Colon cancer is a common type of malignancy in the digestive system. The aim of the present study was to investigate the role of S-phase kinase-associated protein 2 (Skp2) in colon carcinoma and to identify whether depletion of Skp2 by Skp2‑RNA interference (RNAi) attenuates the proliferation and migration of colon carcinoma. Three pairs of small interfering (si)RNA were designed based on the Skp2 gene sequence and the most effective one was used to silence the Skp2 gene in SW620 cells. Subsequent to the interference, quantitative polymerase chain reaction and western blot analysis were used for detecting the expression of Skp-2 mRNA and protein, respectively. The data demonstrated that the Skp2‑siRNA effectively inhibited proliferation (P<0.01), increased the levels of apoptosis and induced G0/G1 phase arrest of the SW620 cells (P<0.01). Transfection of the Skp2 siRNA into SW620 cells effectively reduced Skp2 protein levels, while p27 protein levels increased. In the in vivo experiments, a lentiviral vector of the Skp2-RNAi transfected into SW620 cells markedly inhibited Skp2 expression, as detected by immunohistochemical analysis of nude mice. Additionally, tumorigenicity experiments showed that inhibition of Skp2 significantly increased the survival rate of nude mice. Thus, the in vitro and in vivo results demonstrated that interference of Skp2 expression significantly inhibited the proliferation and induced the apoptosis of SW620 cells. This suggests that Skp2 protein has an important role in the progression of colon cancer. Therefore, Skp2 may enable the early diagnosis of colon cancer and provide new insights into molecular targets for cancer therapy.

摘要

结肠癌是消化系统常见的恶性肿瘤类型。本研究旨在探讨S期激酶相关蛋白2(Skp2)在结肠癌中的作用,并确定通过Skp2-RNA干扰(RNAi)降低Skp2水平是否能减弱结肠癌细胞的增殖和迁移能力。根据Skp2基因序列设计了三对小干扰(si)RNA,并使用最有效的一对来沉默SW620细胞中的Skp2基因。干扰后,分别采用定量聚合酶链反应和蛋白质印迹分析检测Skp2 mRNA和蛋白的表达。数据表明,Skp2-siRNA有效抑制了SW620细胞的增殖(P<0.01),增加了凋亡水平并诱导细胞停滞于G0/G1期(P<0.01)。将Skp2 siRNA转染到SW620细胞中可有效降低Skp2蛋白水平,同时p27蛋白水平升高。在体内实验中,通过对裸鼠的免疫组化分析检测到,转染了Skp2-RNAi慢病毒载体的SW620细胞中Skp2表达明显受到抑制。此外,致瘤性实验表明,抑制Skp2可显著提高裸鼠的存活率。因此,体外和体内实验结果均表明,干扰Skp2表达可显著抑制SW620细胞的增殖并诱导其凋亡。这表明Skp2蛋白在结肠癌进展中具有重要作用。因此,Skp2可能有助于结肠癌的早期诊断,并为癌症治疗的分子靶点提供新的见解。

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