Knockdown of β-catenin by siRNA influences proliferation, apoptosis and invasion of the colon cancer cell line SW480

被引:24
作者
Li, Kui [1 ]
Zhou, Zhong-Yin [1 ]
Ji, Pan-Pan [1 ]
Luo, He-Sheng [1 ]
机构
[1] Wuhan Univ, Renmin Hosp, Dept Gastroenterol, 238 Jiefang Rd, Wuhan 430060, Hubei, Peoples R China
关键词
beta-catenin gene; colorectal cancer; RNA interference; cell proliferation; apoptosis; PATHWAY; DISEASE; TUMOR;
D O I
10.3892/ol.2016.4481
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
The aim of the present study was to explore the effect of knocking down the expression of beta-catenin by small interference (si) RNA on the activity of the Wnt/beta-catenin signaling pathway, and the proliferation, apoptosis and invasion abilities of the human colon cancer cell line SW480. For that purpose, double-stranded siRNA targeting beta-catenin (beta-catenin-siRNA) was synthesized and transfected into SW480 cells. Reverse transcription-polymerase chain reaction (RT-PCR) and western blotting were used to detect the messenger (m) RNA and protein levels of beta-catenin in SW480 cells. To detect cell proliferation, 3-(4,5-dimethylthiazol-2-yl) 2,5-diphenyltetra-zolium bromide assay was performed, while cell apoptosis and caspase-3 activity were detected by flow cytometry and caspase-3 activity assay, respectively. Matrigel invasion assay was performed to detect the influence of siRNA-mediated gene silencing on the invasion and metastasis of SW480 cells in vitro. The results of RT-PCR and western blot analysis demonstrated that, compared with the blank control, negative control and liposome groups, beta-catenin-siRNA transfected SW480 cells had significantly decreased mRNA and protein levels of beta-catenin. In addition, following beta-catenin-siRNA transfection, the proliferation of SW480 cells was significantly lower than that of the blank control, negative control and liposome groups, while the apoptosis rate increased in beta-catenin-siRNA transfected cells, compared with the aforementioned groups. Invasion assay showed that, following beta-catenin-siRNA transfection, the number of SW480 cells infiltrating through the Matrigel membrane was significantly lower than that of the blank control, negative control and liposome groups. Following beta-catenin-siRNA transfection, the caspase-3 activity in SW480 cells was lower than that in the blank control, negative control and liposome groups. These results indicate that siRNA-mediated silencing of beta-catenin could inhibit the proliferation and invasion of SW480 cells and induce apoptosis, thus providing novel potential strategies for the clinical treatment of colon cancer, and may serve as a novel target for cancer therapy.
引用
收藏
页码:3896 / 3900
页数:5
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