| 其他摘要 | Glioma is the most common malignant type in brain cancer, which has poor prognosis and high lethal rate. In these years, the progress in comprehensive therapeutics, including surgery, chemotherapy and radiation therapy, is great; however, the treatment outcome is disappointing because of the high invasive characteristic of the glioma cells, which causes relapse in a short time after treatment. Recently a new concept has been proposed to explain the mechanism of tumorigenesis, which is cancer stem cell (CSC). In this model, a small population of cancer cells is believed to have the potential to self-renewal, unlimited proliferation and multiple differentiations, which are essential for the cancer development and progression. Our study investigated the feasibility to separate the stem-like cancer cells (SLCC) in rat glioma cell line C6 by chemo-reagent colchicine, the cytotoxic effect of the dendritic cell (DC) vaccine pulsed with glioma SLCC proteins against the tumor cells, as well as the preliminary investigation of the genomic instability in SLCC caused by colchicine.
After treatment with different dosages of colchicine (0、0.5、1.0、2.0μg/ml), most cells in rat glioma cell line C6 went to apoptosis, the survival cells expressed stem cell marker Nestin which was assessed by immunoflouresence staining. Furthermore, the results of flow cytometry and RT-PCR show that the expression levels of the stem cell makers, including nestin, sox2 and bmi1, were elevated in SLCC enriched by colchicine as a dosage-dependent way.
DC vaccine is a biological therapy for brain cancer. Our study separated the rat DCs and lymphocytes and expanded in vitro, activated with different SLCC whole protein groups within one week, and then assessed the cytotoxic effect exerted by the activated cytotoxic T lymphocytes (CTL) against cancer cells. The results indicated that the effect of CTL activated by SLCC against cancer cells were stronger than the control groups, which means that glioma SLCC may be a new target for DC therapy.
Besides, the results of RT-PCR also indicated that the expression level of mad2 was elevated after SLCC enrichment, which means that the SLCC enrichment may be caused by the increasing genomic instability in cancer cells, leading to more SLCC in bulk cells.
In summary, our study testified that the SLCC in rat glioma cell line C6 could be separated by chemo-reagent colchicine, and the enrichment might be related to the elevated genomic instability of cancer cells caused by the treatment with chemo-reagent. Meantime, our study also showed that the cytotoxic effect of CTL activated by whole protein from separated SLCC against cancer cells was stronger than the control group. All of these findings provide a preliminary basis for new immunotherapy development targeted the SLCC in glioma. |
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