| ObjectiveGlioma is one of the most common intracranial malignant tumor and it reaches to the 50-60% of the total intracranial tumors. The current treatment of glioma is surgical excision, however, because of the character of invasive growth, it has no clear boarder between normal cerebral tissue and it is hard to excise totally, and the post-surgery recurrence is almost inavoidable. Therefore, the in-depth study on the biological property and growth-controlling factors of the glioma makes a significant and practical role on the post-operational selection of adjuvant chemotherapy and precise prognosis. Our goal based on this problem and aiming at investigating the expression level of apototic inhibitory protein Survivin on the glioma tissue, in order to study the relationship between the expression of Survivin and the heterogenerity of the tumor, to advance the general treating standard of glioma.MethodSubjects: 54 cases of excised human glioma tissue samples were obtained from the patients in charged in the Neurosurgery of the First Affiliated Hospital of China Medical University from January 2003 to December 2004. Classified based on the pathological classification criteria of the human glioma published by WHO (1999), at the same time, set 6 cases of normal tissue as control.Paraffin wrap and slicing with HE stain: After the operation, fixed with 10% Formalin, then, wrapped by paraffin with ensuing 4 micron slicing.Immunohistochemical stain: Normal paraffin slice with dimethylbenzene wash, after that, add 1 drop of peroxide inhibitor to inhibit the endogenous peroxidase activity. Incubate at room temperature for 20 minutes. Put the slice into 0.01mmol/L slightly evaporated 0citrate solution then cool down at room temperature. Add one drop of non-immunized animal serum on individual slices respectively and incubate at room temperature for 20 minutes then discard the serum. Add properly diluted Survivin primary antibody and incubate at 4 deg°C overnight. In the following day, add one drop of biotin marked secondary antibody then incubate at room temperature for 10 minutes. Then, add one drop streotavidin-peroxidase solution and incubate at room temperature for 10 minutes. Add two drops of newly prepared CAB solution to every slice, observe under microscope for 5-10 minutes, then stain with hematoxylin. After that, fix the slice with resin. The Semi-quantitafication standard, naked-eye observing and automatic graphic analysis system exam the optical density of a specified field. Processing the acquired data with SPSS 10.0 for Windows statistic software.ResultThe results of the staining all showed the differential expression of Survivin in glioma tissue, however, in the normal cerebral tissue, the Survivin expression is negative. And the expression levels of Survivin have obvious differences, the higher grade of the tumor, the more the Survivin expression, and with statistic significance (p<0.01). At the same time, glioma in different grades have different optical density of the Survivin expression, the higher the tumor, the higher the optical density of the expressed Survivin. Malignant glioma has heterogneity and we compared the optical density of the Survivin in central and peripheral portion of the tumor, they have an obvious difference (p<0.001).ConclusionSurvivin in normal cerebral tissue is negative but expressed in malignant glioma tissue and increased with the increased pathological grade of the tumor which suggests the quantitative evaluation of Survivin could be set as a criterion for pre-operational malignancy evaluation and prognosis.The survivin expression in a single tumor body is higher than the peripheral edematous region and is a reflexion the heterogeneity and invasion region of a tumor. It is significant to the proposed area of surgical excision.
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