| Summary: | 博士 === 長庚大學 === 臨床醫學研究所 === 90 === Functional inactivation of TSG101 in mouse fibroblasts results in cell transformation and the ability to form metastatic tumor in nude mice. The human tsg101 mapped to chromosome 11q15.1-2 and found to mutate in some human malignancy.
To evaluate the expression pattern in Taiwan breast cancer patients, we analysed the mRNA, protein, and the protein location of tsg101 in breast cancer specimens. RT-PCR data showed a truncated transcript was frequently expressed in late stage breast cancer specimens. TSG101 protein was mainly located in cytoplasm, and cell nuclei was occasionlly immunopositive and the chromosomes were deeply stained during cell division.
To investigate the tsg101 truncated forms, we cloned these truncated forms and the sequencing results showed that the B7 was composed of three type truncations. Overexpression of these tsg101 truncated forms in breast cancer cell line using adenovirus delivery system will induced cell cycle arrest before M phase.
We generate N-terminal anti-tsg101 truncated form polyclonal antibody, and tagging epitope to these tsg101 truncated forms with HA, flag, and GFP in N- and C-terminal of tsg101 truncated forms. Western blotting suggesting that tsg101 truncated forms were easily degraded.
Overexpression of tsg101 truncated forms in 293 cells using gene transfer method was harmful to cells as compared with the control group. Fluorescent microscopy showed that the location of these tsg101 truncated forms were also different from GFP control group.
Key words: Adenovirus, breast cancer, growth arrest, tsg101
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