Experimental Studies on Therapy of Lung Cancer by VEGF-C siRNA Mediated by Liposomes
|Course||Department of Cardiothoracic Surgery|
|Keywords||Vascular endothelial growth factor C (VEGF-C) RNA interference Lung cancer Gene therapy|
Objective: Lung cancer is the high incidence of respiratory malignancies, usually vivo gene mutation leads to changes in cell cycle and apoptosis inhibition. In recent years, many experimental studies have shown that VEGF-C plays an important role in tumor development. In this study, through in vitro synthesis of small molecules targeting VEGF-C mRNA interference RNA (short interfering RNA, siRNA) liposome-mediated transfection of human lung cancer A-549 cell line, observed VEGF-C mRNA and VEGF-C protein expression and cell growth and apoptosis. Method: the design and synthesis of VEGF-C gene sequence specific small interfering RNA by liposome-mediated transfection of lung cancer A-549 cells to silence the expression of VEGF-C. Transfection efficiency was observed with a fluorescence microscope and inverted microscope and morphological changes after transfection; siRNA and liposome detection experiment concentration of pure observation targeting VEGF-A-549 cells inhibited by MTT assay method, C siRNA via liposome conductor infected lung adenocarcinoma A-549 cell growth inhibition and cell growth curve. Using semi-quantitative reverse transcriptase polymerase chain reaction (reverse transcriptase polymerase chain reaction, RT-PCR) and protein immunoblot method (by western blotting) Determination of VEGF-C mRNA and protein expression. VEGF-C siRNA A-549 cell apoptosis was detected by flow cytometry. Transmission electron microscopy to observe changes in cell ultrastructure of exogenous VEGF-C siRNA transfected A-549 cells. Results: liposome-mediated siRNA transfected lung adenocarcinoma A-549 cells. Experimental concentrations of pure siRNA and liposomes of A-549 cells without significant toxicity. Liposome-mediated VEGF-C siRNA transfected lung cancer cells after 48h, the experimental group cells adherent infiltration decline in their ability to grow, some cells off shrinkage, round cell morphology, reduced in size, refraction enhanced blank control group and the experimental control group of normal cell growth. VEGF-C gene sequence specific siRNA can effectively inhibit VEGF-C mRNA and protein expression in lung adenocarcinoma A-549 cells, and the differences were statistically significant (P lt; 0.05) compared with the control group and the experimental control group. Down the expression of VEGF-C in vitro significantly inhibit the growth and proliferation of A-549 cells in a concentration-dependent inhibition rate within a certain range of concentrations, lowered VEGF-C A-549 can induce apoptosis increase 48h after transfection, the experimental group growth inhibition rate (38.87%), the rate of apoptosis (19.31 ± 2.21)% was significantly higher than that in the control group (P lt; 0.05), typical of apoptotic cells after transfection with siRNA and found under a transmission electron microscope, the nuclear chromatin PACK, uneven distribution. Conclusion: RNA interference technology can effectively inhibit VEGF-C mRNA and protein expression of VEGF-C and lower lung adenocarcinoma A-549 cell proliferation, induction of apoptosis increased, providing theoretical support for the targeting of VEGF-C siRNA treatment of lung cancer, RNA interference as a treatment for lung cancer means worthy of further research and promotion.