Vascular endothelial growth factor (vegf)

The paraffin sections were heated in an oven. After hydration, they were treated with eosin staining for 1 minute and hematoxylin for 30 seconds. As for immunofluorescence staining, the hydrated sections were first treated by citrate-based antigen unmasking solutions (Vector Laboratory, America). They were incubated with antibodies (1 : 100) against PCNA (Solarbio, China) and VEGF (Solarbio, China), respectively. The sections were then incubated with commercial secondary antibody (1 : 200). The sections were photographed with Nikon microscopy (Nikon, Japan).

SLC27A1 (Solarbio, China, dilution:1:200), PTBP1 (Solarbio,China, dilution:1:100), EIF5A (Solarbio, China, dilution:1:50), Ki67 (Solarbio, China, dilution:1:100), VEGF (Solarbio, China, dilution:1:100), and P53 (Solarbio, China, dilution:1:200) were evaluated by immunohistochemistry. The paraffin-embedded specimens were cut into 4 μm thick sections. According to the manufacturer’s instructions, an antibody immunohistochemical analysis was performed on all sections using Leica Bond-III's automatic, random, and continuous slide staining system (Leica Biosystems, Germany). Images were obtained using a whole-slide scanner (3DHISTECH Ltd, Budapest, Hungary) and the immunohistochemical expression was scored by two observers using the CaseViewer software. The criterion of immunohistochemistry was that there were obvious brown granules in the cytoplasm or nucleus. The H score (range 0–300) was obtained by multiplying the staining intensity (0–3) by the percentage of positive cells (0%–100%), and the expression of tumor cells was scored semi-quantitatively.