科研之友
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摘要
Aims : Dual gene targeting siRNA (DGT siRNA) composed of Neuroepithelial transforming gene-1 (NET-1) and Vascular endothelial growth factor (VEGF) siRNA sequences was used to silence NET-1 and VEGF gene in lung cancer (LC) cell. And then the biological characteristics of LC cell were investigated. Materials and methods : DGT siRNA, NET-1 and VEGF was transfected to lung adenocarcinoma cells lines A549. Quantitative real-time PCR (RT-qPCR), Western Blot immunofluorescence was used to detect the transcription and expression of NET-1 and VEGF gene. Cell proliferation was detected by MTT (methyl thiazolyl tetrazolium) assay. Apoptosis were evaluated using Annexin V-FITC (Annexin V-Fluorescein Isothiocyanate) assay and Hoechst staining. The migration invasion potential of A549 cells were tested by Wound healing and Transwell invasion experiment. Supernatant of A549 with or without treated by siRNA was applied to incubate the HUVECs (human umbilical vein endothelial cells). The tube formation ability of HUVECs was investigated. Results: Single targeting siRNA and DGT siRNA could down-regulate the expressions of NET-1 and VEGF mRNA and protein expression. In DGT siRNA transfected group, the down-regulated gene expression of NET-1 and VEGF was more obvious. The apoptosis rate was increased and the ability of migration and invasion was decreased after transfection by single or DGT siRNA. The effects in DGT siRNA transfected group were more obvious, as well. Concentration of VEGF in the supernatant obtained from A549 cells transfected by single and dual targeting siRNA decreased and HUVECs tube formation ability was decreased compared with the control group. Conclusions: Biological behavior and angiogenesis of lung cancer can be inhibited by NET-1 and VEGF silencing technology. Simultaneous silencing of NET-1 and VEGF using DGT siRNA achieved better inhibiting effects, which may provide an alternative therapeutics for LC.
- 出版日期2015
- 单位南通大学
