| Summary: | 碩士 === 國立成功大學 === 藥理學研究所 === 92 === Our lab had previous shown that cold water extract from a local variety of Canavalia gladiata, termed IC-1, possessed some interesting anticancer activities. In this investigation, I clarify and identify further the anticancer activities of IC-1. IC-1 shows in vitro cytotoxicities toward some cancer cell lines in a dose-dependent manner. Flow cytometry assay reveals that cancer cells death is due to apoptosis, which is supported by two more assays—DNA fragmentation and Hoechst 33258 nucleus staining. Gelatin zymography of matrix metalloproteinase 2 and 9, the two indicators of tumor invasion of basemembrane, indicates that IC-1 can inhibit both enzymes. Next I tested the antiangiogenesis of IC-1 using bovine aorta endothelial cell (BAEC) in a migration assay. The result indicates IC-1 can inhibit the blood endothelial cell migration. Furthermore, I tested the anticancer activity of IC-1 in vivo using ML1-4A hepatoma cells as primary tumor model. IC-1 treatment reduces tumor size by more than 50% as compare to the control, confirming further the previous observation made in this lab.
The immune boosting effect of IC-1 was confirmed and further analyzed in vivo. Injected intraperitoneally into Balb/c mouse, IC-1 was found to stimulate splenocytes proliferation, which was assayed by 3H-thymidine incorporation method. The proliferated cell population, identified by fluorescence activated cell sorter (FACS), was found to be B cells, T cells and nature killer cells. IC-1 also can stimulate the expression of cytokines IL-2 and IFN-g. In addition, IC-1 can reverse the immune impairment, including B cells, T cells and nature killer cells in the spleen, elicited by cisplatin.
Taken together, these results prove that IC-1 not only inhibit tumor growth but also activate immune responses, a dual effect that renders its anticancer activities.
|
|---|
