Journal: International Journal of Oncology
Article Title: VEGF-D-enhanced lymph node metastasis of ovarian cancer is reversed by vesicular stomatitis virus matrix protein
Figure Lengend Snippet: RT-PCR and apoptotic effect on VEGFD-SK cells. (A) RT-PCR analysis of expression of MP in vitro . High expression of MP in VEGFD-SK cells transfected with pVAX-MP:lip was detected (1,036 bp). GAPDH was used for the internal standard. The displaying panel of electrophoretic image presents the following: DNA ladder marker (a), WT-SK (b), EV-SK (c), VEGFD-SK (d), pVAX:lip (e), and pVAX-MP:lip (f), respectively. (B) Overexpression of VEGF-D using RT-PCR could be detected in VEGFD-SK cells, but not in WT-SK and EV-SK cells (1,077 bp). However, the VEGF-D expression was suppressed when VEGFD-SK cells suffered from pVAX-MP:lip. The overexpression of VEGF-D did not decreased obviously when VEGFD-SK cells were treated with pVA. (C) VEGF-D expression in vivo was analysis by RT-PCR. (D) The expression of VEGF-D in vivo was determined with immunohistochemistry using VEGF-D antibody: VEGFD-SK tumors were detected with much stronger staining. Whereas, the VEGFD-SK tumors in pVAX-MP:lip group present nearly negative staining. Tumors in pVAX:lip group also exhibited strong staining. Very weak VEGF-D protein was observed in WT-SK and EV-SK tumors. (E) Hoechst-33258 stained fluorescence microscopy (x200) in vitro : apoptosis cells were observed apparently when the VEGFD-SK cells were treated with pVAX-MP:lip (arrow). (F) A quantitative comparison of apoptotic cells was carried out by flow analysis stained with Annexin V-FITC and propidium iodide: pVAX-MP significantly increased VEGFD-SK cells apoptosis compared with the controls.
Article Snippet: Apoptosis analysis Qualitative apoptosis was determined by Hoechst-33258 compounds (Sigma) according to the manufacturer's instructions.
Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, In Vitro, Transfection, Marker, Over Expression, In Vivo, Immunohistochemistry, Staining, Negative Staining, Fluorescence, Microscopy, Flow Cytometry