| Summary: | 碩士 === 國立成功大學 === 生物化學研究所 === 83 === We want to study how to use gene therapy on bladder tumor. The
effective rate for gene therapy is usually about 30-40%. The
reason for the variation of gene therapy efficiency may partly
result from the individual expression pattern of oncogenes
and tumor suppressor genes in different tumors. Overexpression
of oncogene neu has been observed in 68% of bladder tumors,
therefere , we wanted to investigate the role of neu in bladder
cancer development. 1, normal rat neu cDNA was delivered into
MBT2 cell to generate MBT2-NEU. The MBT2 cells can be
transplanted into syngeneic mouse C3H/HeNcrj to study immune
response against tumor. 2, defective retrovirus which contain
human interleukin 2(hIL2) gene was used to infect MBT2 and MBT2-
NEU,and to generate MBT2-IL2 and MBT2-NEU-IL2 cells. These cell
lines that could secrete IL2 were irradiated with.gamma.-ray
and injected into C3H mouse.Overexpression of neu in MBT2-NEU
transfectants N18 and N19 was demonstrated by western blot. The
control transfectants N21 and NC that contained no pSVneu
integration were selected with hygromycin and isolated. The N18
and N19 transfectants showed weaker tumorigenicity than control
transfectants and parental cells in syngenic C3H mice or SCID
mice. On the other hand, there are no significant difference in
soft agar assay and morphology observation among these cell
lines .MBT2, MBT2-NEU, MBT2-IL2, and MBT2-NEU-IL2 in a amount
of 500000 cells with or without 6000 Rad .gamma.ray irradiation
were transplanted in C3H mouse and assayed for tumor formation.
Mice inoculated the irradiated cell lines were further injected
with MBT2 or MBT2-NEU cells observe the antitumor effect by the
first vaccination with irradiated tumor cells. Mice treated
with irradiated MBT2-IL2 or MBT2-NEU-IL2 did not develop any
tumor.The result indicated that the tumor vaccine generate the
immune protection response against futher transplant of tumor
cells.
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