Functional analysis of p130 oncogene in malignant glioma

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I. Analysis of cell cycle and cyclin with use of FCMThree kinds of glioma cell lines were analyzed the cell cycle and expression of cyclins from the DNA histogram stained with FTTC laelled cyclin A, B1, D1, E and propidium iode. These cell lines experssed all of cyclin A, B1, D1. Cyclin B1 was prooved to be expressed on the S, G2, M phases and cyclin D1 was expressed on the phase of G0, G1 phases.II. Analysis of cell cycle and cyclin with use of LSCSame results were obtained by LSC as well as FCM.III. Effect of As2O3 on cell cycle progression and cyclin expression in glioblastoma cell linesAs2O3 induced an increase in p53 level and a decrease in level of cyclin B1, combined with cell arrest at G2/M in cell lines. Cell arrest in G1, however, was associated with a decline in cyclin D1 expression in the U87 MG cells.IV. Rb2/p130 expression on the surgical specimen of gliomasBenign gliomas grade I and II were expressed strongly, but malignant glioma grade IV showed negative stain. On the other hand, p53 showed high positive rates in the cases of grade IV.V. Functional analysis of p130 induced gliomap130 cDNA induced subclone to BCMGSneo and this p130 was transfected into U87 MG with use of Lipofact AMINE. After transfected cells were sellected by G418, U87 MG expressed of p130 oncogene was success to be produced. Plasmid from the oncogene of p130 was obtained over 6 months and p130 was possible to be transofected into U87MG cell line. We are planning now how to controll the proliferation of malignant gloma, especially mutant glioma (N-319) or express of the cyclins.