Effects of ephrinB2 gene siRNA on the biological behavior of human colorectal cancer cells

Li,Peiwu; Chen,Wangsheng; Wang,Yingzhen; Fu,Xu; Wen,Kunming; Qian,Jiang; Huang,Chun; Fu,Zhongxue

doi:10.3892/or.2014.3633

Effects of ephrinB2 gene siRNA on the biological behavior of human colorectal cancer cells

Authors:
- Peiwu Li
- Wangsheng Chen
- Yingzhen Wang
- Xu Fu
- Kunming Wen
- Jiang Qian
- Chun Huang
- Zhongxue Fu
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Affiliations: Department of General Surgery, First Affiliated Hospital, Chongqing Medical University, Chongqing 400010, P.R. China, Department of Emergency, Lanzhou University Second Hospital, Lanzhou 730000, P.R. China
Published online on: November 27, 2014 https://doi.org/10.3892/or.2014.3633
Pages: 758-766

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Abstract

Colorectal cancer (CRC) is a common gastrointestinal malignancy worldwide and is a lethal and aggressive malignancy with a dismal prognosis. EphrinB2 is a membrane-bound ligand and has an intracellular domain that also possesses an intrinsic signaling capacity called ‘reverse signaling’. In the present study, CRC cell lines were screened for high expression of ephrinB2. Small interfering RNA (siRNA) knockdown of ephrinB2 was performed in human SW480 CRC cells. The levels of expression of ephrinB2, VEGF, CD105 and matrix metalloproteinase 9 (MMP9) protein were measured by western blotting, and messenger RNA (mRNA) levels were measured using real-time PCR. Apoptosis and cell cycle distribution were determined using flow cytometry. Cell proliferation was measured by a methyl thiazole tetrazolium (MTT) test and a scratch healing experiment was used to measure the extent of cell migration. A Transwell assay was used to detect the extent of cell invasion. The results showed that RNA interference (RNAi) of ephrinB2 effectively silenced the ephrinB2 gene at both the mRNA and protein levels in SW480 cells and inhibited the proliferation, invasion, migration and angiogenesis and induced apoptosis in SW480 cells. These effects may be attributed to VEGF and MMP9 regulation.

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