The effect of RNA interference-mediated down-regulation of EphB4 on the growth of malignant glioma cell line U251 / 肿瘤

ABSTRACT

Objective: To determine the interfering effects of EphB4-targeted short interfering RNA (siRNA) on EphB4 mRNA expression and its effect on the growth of glioma U251 cell line. Method: EphB4-targeted siRNA was designed and synthesized, and then was transfected into U251 cells. The inhibition of EphB4 mRNA expression was detected by RT-PCR. The effect of EphB4-targeted siRNA on cell growth rate was measured by CCK-8 method. Cell apoptosis was tested by flow cytometry (FCM). Wound healing test was used to observe the migration ability of cells. The invasiveness of tumor cells was evaluated by counting the number of cells passing the Transwell membrane. Results: EphB4 mRNA transcription level was decreased by 75.0% after transfection of malignant glioma U251 cells with 100 nmol/L siRNA-EphB4. The inhibition of cell proliferation was in a dose-dependent manner. FCM analysis showed that cells were arrested at sub-G1 phase at different degrees and the migration capacity decreased after transfection with 100 nmol/L siRNA-EphB4 compared with the negative control. The number of cells permeating the matrigel membrane significantly were decreased in the siRNA-EphB4 transfection group compared with the control group. Conclusion: siRNA-EphB4 markedly targetes and knocks down EphB4 gene transcription. Down-regulation of EphB4 affects cell proliferation and induces apoptosis of cells. Transfection of siRNA-EphB4 into U251 cells inhibits the migration and invasion abilities of cells at various degrees. It indicates that silencing EphB4 expression might become a noval approach in the treatment of glioma.