Study On The Correlation Of The Malignance And Angiogenesis Of Astrocytic Tumors Among HGF, C-Met And VEGF

ObejectiveTo detect the microvessel density (MVD) in astrocytic tumors. To detect the correlation between the expression of HGF, C-met and VEGF and the pathological classification and MVD of astrocytic tumors. And then preliminarily discuss the relation of the extent of expression of HGF, C-met and VEGF in astrocytic tumors with their clinicopathological parameters.Materials and Methods93 patients suffering from astrocytic tumors with complete clinical data who received first surgical removal in Shandong Provincial Hospital between 2008 Jan and 2010 May were selected, including 53 male and 50 female patients, whose average age was 38.33±13.06. With hematoxylin-eosin (HE) staining, they were classified as low-level (Level II,28 cases) and high-level (Level III&IV,65 cases) which included 72 supratentorial tumors and 21 infratentorial ones, according to 2000 WHO neuroglioma staging.9 patients suffered from traumatic brain injuries received intracranial decompression were selected as control group.S-P immunohistochemistry (IHC) method was employed to detect the expression of HGF, C-met and VEGF protein, proceeding according to standard method.5 visual fields were chosen randomly to count positive cells under high magnification. Positive cells were defined as ones with brown or tan particles in cell plasma and without background staing. They were also classified according to the intensity of staining and the number of positive cells:negative (-):no positive cells; weak-positive (+):positives cells accounting for less than 30% of the total cells; positive (++):positives cells accounting for 30-60% of the total cells; strong-positive (+++):positive cells accounting for more than 60% of the total cells.S-P IHC was employed to determine MVD:count vascular endothelial cells stained brown under high magnification (400×) in 5 visual fields randomly, and MVD was defined as average cell numbers.RNA was extracted and reverse-transcripted according to standard methods. Then we determined the RNA levels of HGF, C-met and VEGF with Real-time PCR. We selected (3-actin as internal parameters. Primers targeting genes were designed and synthesized by TAKARA Biology Company according to their corresponding sequences in GenBank. The quantity of target genes was obtained in terms of relative quantification.Statistical analysis:Spearman Rank Correlation Analysis was employed to analyze the correlations among MVD, gender, age and the protein expression of HGF, C-met and VEGF. Pair-matching t test was employed to determine the relationship between normal brain tissues and astrocytic tumors in terms of the RNA level of HGF, C-met and VEGF. We employed Linear Correlation Analysis to detect the reciprocal correlation among HGF, C-met and VEGF. P values <0.05 were considered to be statistically significant. All statistical process used SPSS 13.0 software.Results:1. Protein expression of HGF, C-met and VEGF in astrocytic tumors tissuesThe protein expression of HGF, C-met and VEGF in variant levels of astrocytic tumors tissues were higher than normal brain tissues (P<0.01), and the positive extents correlated with the pathological levels (P<0.01) and MVD (P<0.01). No statistical significance was found between positive extents and age (P=0.78) and gender (P=0.67). The reciprocal correlation among HGF, C-met and VEGF had statistical significance.2. The mRNA levels of HGF, C-met and VEGF in astrocytic tumorsThe mRNA levels of HGF, C-met and VEGF in astrocytic tumors were higher than those in normal brain tissues (P<0.01). The mRNA levels of HGF, C-met and VEGF rose in accordance with the pathological levels of astrocytic levels. Statistical significance was found in the correlation among the mRNA levels of HGF, C-met and VEGF in astrocytic tumors.Conclusion1. There is high expression of HGF, C-met and VEGF in astrocytic tumors. 2. There is a statistically significance in HGF, C-met and VEGF expression in astrocytic tumos of variant pathological levels. HGF, C-met and VEGF are highly associated with the malignance of astrocytic tumors.3. HGF, C-met and VEGF may serve as useful tools for confirmation of astrocytic tumors; HGF, C-met and VEGF genes may also indicate potential targets for astrocytic tumors gene therapy.