Functional analysis of malignant brain tumor related gene <h-neu> and development of a novel molecular tageting therapy based on the h-neu gene function

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We have been investigated about the analysis of the function in the product of a novel gene < h-neu>. We also have tried several experiments to investigate how this gene prduct functions in the malignant glioma cells. First, we made a new antibody to recognize the epitope of the h-neu protein by ourselves. Since we could confirm that this antibody worked in immunhistochemistry, we tried to stain the tumor samples in every grade of gliomas. However, we could not find out the significant differences in the sensitivity of the staining, therefore, we concluded that the expression level of h-neu in glioma was not so high. Next, we performed a Real-Time PCR in every grade of glioma samples. The expression of the mRNA of h-neu in malignant glioma was low compared as those of low grade gliomg. To investigate how the reduction of the product of h-neu affect the growth rate of malignant gliomas, we made a siRNA of h-neu and performed the transfection of it to several glioma culture cells. The reduction of the growth rate due to the transfection of h-neu siRNA was various in glioma cell lines. We considered that the variety of the reduction rate of growth in glioma cell lines was basically due to the native expression of h-neu in each cell lines, therefore, we performed the Nothern blot analysis and measured the expression level of h-neu in each cell lines. However the there was no relation between the expression of h-neu and the reduction rate of the growth due to siRNA in glioma cell lines. We considered that we should attempt to investigate the function of h-neu protein, we have started the functional analysis of h-neu using the proteomix procedure. We are now preparing the glioma cell line in which h-neu was overexpressed. Furthermore, we are still planning several experiments to investigate the h-neu function. We would like to know the normal function of h-neu and after that we would like to try to know the meaning 3 of h-neu function in glioma cell lines.