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Application of Trichoderma pseudokoningii extracellular polysaccharides having anticancer activities

A Trichoderma cell, Kang's technology, applied in the field of medicine

Active Publication Date: 2012-09-12
博德生物技术(德州)有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0006] At present, the functional research on the exopolysaccharide of Trichoderma pseudokonii has become a hotspot in the research of Trichoderma pseudokonii, but the research on the effect of the exopolysaccharide on the growth of human chronic myeloid leukemia cells K562 has not been seen. to report

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  • Application of Trichoderma pseudokoningii extracellular polysaccharides having anticancer activities
  • Application of Trichoderma pseudokoningii extracellular polysaccharides having anticancer activities
  • Application of Trichoderma pseudokoningii extracellular polysaccharides having anticancer activities

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Embodiment 1

[0042] A method for preparing exopolysaccharide from Trichoderma pseudoconii with anticancer activity, comprising the steps of:

[0043] (1) inoculate Trichoderma pseudokoningii (Trichoderma pseudokoningii) with strain preservation number CGMCC No.1443 in PDA medium, and activate it at 28°C for 24 hours to obtain activated mycelium;

[0044] The PDA medium has the following components per liter: 200g of potatoes, 20g of sucrose, 2g of agar, water to 1000mL, natural pH;

[0045] (2) Transfer the activated hyphae prepared in step (1) to the PDB fermentation medium, culture at a temperature of 28° C., and culture on a shaker at 120 rpm for 10 days to obtain a liquid fermentation broth;

[0046] The PDB fermentation medium has the following components per liter: 200g of potatoes, 20g of sucrose, water to 1000mL, natural pH;

[0047] (3) Filter the liquid fermented liquid prepared in step (2) through 4 layers of gauze to remove mycelia, and concentrate the filtrate to 25% of the o...

Embodiment 2

[0057] Effects of EPS-1 Administration in Vitro on the Proliferative Activity of Human Chronic Myelogenous Leukemia Cell K562

[0058] SRB is a protein-binding dye that can combine with the basic amino acids of cellular proteins fixed by trichloroacetic acid to present a bright pink color, and its color change is proportional to the protein in living cells. ELISA The absorbance is measured by the detector to obtain an accurate value of cell protein content, which can be used to calculate the number of cells and reflect the proliferation activity of cells.

[0059] experimental method

[0060] Human chronic myelogenous leukemia cells K562 in the logarithmic growth phase were taken to make a density of 1×10 4 cells / ml cell suspension, seeded in 96-well plate, 180 μL / well. After culturing for 24 hours, add 20 μL of exopolysaccharide solution of Trichoderma pseudoconii in different concentrations to each well (final concentration is 0.2, 0.4, 0.6, 0.8, 1 mg / ml), the total liquid...

Embodiment 3

[0064] Effects of exopolysaccharide from Trichoderma pseudoconii in vitro on the chromatin morphology of human chronic myelogenous leukemia cells K562

[0065] When cells undergo apoptosis, the permeability of the cell membrane changes, and the chromatin condenses and produces apoptotic bodies. After staining with Hoechst33258 staining solution that has membrane permeability and can specifically bind DNA, it can be visualized under an inverted fluorescence microscope. Morphological changes of the nuclei were observed.

[0066] experimental method

[0067] Place sterile coverslips in a six-well plate, seed human chronic myelogenous leukemia cells K562 and incubate for 24 hours. The next day, different concentrations of exopolysaccharide solutions of Trichoderma pseudoconii were added to stimulate the cells for 48 hours, the culture medium was removed, 0.5ml of fixative was added, and the cells were fixed for 10 minutes. Blot off the fixative, add 0.5ml Hoechst 33258 staining ...

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Abstract

The invention relates to an application of Trichoderma pseudokoningii extracellular polysaccharides having anticancer activities. The Trichoderma pseudokoningii extracellular polysaccharides having the anticancer activities can reduce the vitality of K562 cells in vitro when they are applied to tumor growth inhibition (auxiliary) medicines, so intracellular active oxygen and free calcium ion concentrations are improved, the expression abundance of mRNA of p53 and Bax genes is improved, the expression of the Bc1-2 transcription level is reduced, the ectropion of cell membrane phosphatidylserine of the human chronic granulocyte leukemia cells K562 is induced, and the nuclear polycondensation and the fragmentation of K562 are induced to generate apoptotic bodies and apoptosis; and in-vivo administration can increase weights of tumor bearing mice and alleviate growth speeds and weights of tumors of the tumor bearing mice, thereby tumor cell killing and tumor growth inhibiting purposes are reached.

Description

technical field [0001] The invention relates to the application of Trichoderma pseudoconii exopolysaccharide with the activity of inhibiting the growth of K562 cells, and belongs to the technical field of medicine. Background technique [0002] According to the World Health Organization statistics, as of 2009, the global average life expectancy has reached 68 years. However, electromagnetic pollution in modern society, heavy metal pollution in soil and water, air pollution, etc. have caused more and more harm to human health. With the aging of the global population and the continuous growth of the world's total population, as well as the increase in high-risk behaviors for cancer (especially smoking in developing countries or regions), the number of cancer patients worldwide has increased dramatically. Statistics from GLOBOCAN in 2008 showed that among the approximately 12.7 million cancer cases and 7.6 million cancer deaths, about 56% of the cancer cases and 64% of the can...

Claims

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Application Information

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IPC IPC(8): A61K31/715A61P35/02C12P19/04C12R1/885
Inventor 陈靠山陈国创张鹏英黄涛涛
Owner 博德生物技术(德州)有限公司
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