Method for treating cancer
a pancreatic cancer and epidermal gel technology, applied in the field of catfish epidermal gel secretions, can solve the problems of many side effects, high cost of conventional drugs for treating pancreatic cancer, and toxic to the blood and other internal organs of many conventional drugs
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example 1
Preparation of SPF and Calculation of Soluble Protein in Solution
[0048]EGS was collected from the catfish skin and kept at −80° C. until use. Frozen EGS was thawed to 4° C., mixed with an equal volume of extraction buffer [phosphate buffered saline (PBS), 0.05 M containing (NaH2PO4 / Na2HPO4) and 0.14 M NaCl, pH 7.5], and homogenized with an Ultra Truex (IKA) homogenizer. This step and all subsequent purification procedures were carried out at 4′C unless otherwise indicated. The homogenate was centrifuged at 15,000 rpm for 15 minutes. The supernatant was collected, and the pellet (insoluble protein etc.) was re-extracted with extraction buffer (2-4 times for different exemplary compositions). Each time, the soluble fraction was separated by centrifugation as described above, and the two extracted fractions were pooled. The combined extracted fractions provided the soluble protein fraction (SPF) (Fraction B).
[0049]To find the concentration of catfish soluble proteins in the SPF (Fracti...
example 2
Effect of Fraction B on Human and Mouse Pancreatic and Liver Cancer Cells
[0051]To understand whether fraction B has the ability to inhibit proliferation of human cancer cells, especially pancreatic cancer and liver cancer, the effect of Fraction B in the proliferation of mouse Panc-02 cells and human pancreatic cancer Panc-1 and BxPC3 cells as well as human liver cancer Hep3B cells was studied. Intriguingly, all four cell lines responded to Fraction B treatment dose-dependently with very similar potency with IC50 for all four cell lines being at 200-400 μg / kg. The results suggested that the anti-proliferative effect of Fraction B in human pancreatic and liver cancer cells is mediated through similar mechanisms (FIGS. 1A-1C).
example 3
Determining Whether Fraction B Inhibits Growth of Panc-01 Cells and Panc-02 Cells Through Apoptosis
[0052]To determine whether the inhibitory effect of Fraction B upon cancer cell growth is mediated through reducing cell proliferation or induction of apoptotic cell death, cell cycle and apoptotic cell death in Fraction B treated Panc-1 and Panc-02 cells were measured by propidium iodine staining. As shown in FIGS. 2A and 2B, Fraction B (500 μg / ml) significantly increased subG1 / G0 phase cells by 2-fold and 25 fold in Panc-1 cells and Panc-02 cells respectively, vs. vehicle treatment, suggesting that the cells were undergoing apoptosis.
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