Application of hericium erinaceus polysaccharide in preparation of product for preventing and treating colon cancer
A technology of Hericium erinaceus polysaccharides and colon cancer, applied in the field of medicine, can solve the problems of colon cancer without research and reports, and achieve the effect of inhibiting the ability of clonal colony formation and promoting apoptosis
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Embodiment 1
[0058] Embodiment 1, the impact of Hericium erinaceus polysaccharide on the survival rate of human colon cancer cells
[0059]MTT method was used to study the inhibitory effect of Hericium erinaceus polysaccharides on the viability of HCT-116, DLD1, HT-29 cells. Human colon cancer cells HCT-116, DLD1, and HT-29 in the logarithmic growth phase were digested with 0.2% trypsin, seeded in a 96-well plate at a cell density of 6000 cells / 100 μL, and incubated at 37°C in 5% CO 2 Incubate for 24 h in an incubator.
[0060] After the cells adhered to the wall, fresh medium was replaced, and different concentrations of Hericium erinaceus polysaccharides were added, and 5 replicate wells were set for each concentration, and a blank control group was set at the same time, and the incubation was continued for 48 hours under the same conditions. Aspirate off the old medium, wash twice with PBS, add 100 μL of fresh medium to each well, then add 20 μL of MTT, continue to incubate for 4 hours...
Embodiment 2
[0062] Example 2. Effect of Hericium erinaceus polysaccharides on the clone of human colon cancer cell HCT-116 cells.
[0063] The effect of Hericium erinaceus polysaccharide on the proliferation ability of HCT-116 cells was further verified by the colony formation experiment of cell clones.
[0064] HCT-116 cells in logarithmic growth phase were selected, digested with 0.2% trypsin, and seeded in a 6-well cell culture plate at a cell density of 1000 / well, 37°C, 5% CO 2 Cultured in an incubator for 24 hours; after the cells adhered to the wall, change the fresh medium, add different concentrations of Hericium erinaceus polysaccharides, set 3 replicate wells for each concentration, and continue to incubate for 6 days under the same conditions. The medium in each well was discarded, washed three times with PBS, the cells were fixed with immunostaining fixative for 1 h at room temperature, stained with 0.1% crystal violet for 30 min, and the formation of HCT-116 cell clones was o...
Embodiment 3
[0066] Embodiment 3, cell apoptosis detection
[0067] The HCT-116 cells were treated with 0, 0.3, 0.6 mg / ml Hericium erinaceus polysaccharides for 48 hours, and the effect of Hericium erinaceus polysaccharides on cell apoptosis was detected by Annexin V / PI double staining method.
[0068] The results showed that the apoptosis rate of HCT-116 cells was 16.03±1.32% and 29.75±0.96% after treatment with 0.3 and 0.6mg / ml Hericium erinaceus polysaccharides, confirming that Hericium erinaceus polysaccharides can induce colon cancer HCT-116 cells to undergo apoptosis. death (such as image 3 A, 3B, 3C, image 3 A, 3B, 3C are flow cytometry detection of 0, 0.3, 0.6 mg / ml Hericium erinaceus polysaccharides induced apoptosis in human colon cancer HCT-116 cells; image 3 D represents the effect of Hericium erinaceus polysaccharide on the apoptosis rate).
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