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Preparation of fermentation liquor for inhibiting liver cancer cell growth

A technology for inhibiting liver cancer and fermentation broth, applied in the field of preparation in the field of biotechnology, can solve the problems of isolating endophytic fungi with anti-tumor effect, poor systemic chemotherapy effect of liver cancer patients, early metastasis of cancer cells, etc., and achieves good development prospects, Good cancer cell selectivity and avoid the effect of poor water solubility

Inactive Publication Date: 2010-09-29
SHANGHAI JIAOTONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Liver cancer is a disease with a high mortality rate in the world. The general treatment method is surgical resection. However, in many cases, it occurs in the center, and early metastasis of cancer cells is very easy to occur. This condition limits the application of surgical resection.
Therefore, adjuvant therapy with some drugs is needed, but the effect of existing drugs on systemic chemotherapy for patients with liver cancer is poor, so finding new effective chemotherapy drugs is of great significance to improve the survival rate of patients with advanced and recurrent liver cancer
However, there is no report on the isolation of endophytic fungi with antitumor effects from Juglans catalpa

Method used

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  • Preparation of fermentation liquor for inhibiting liver cancer cell growth
  • Preparation of fermentation liquor for inhibiting liver cancer cell growth
  • Preparation of fermentation liquor for inhibiting liver cancer cell growth

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Isolation of endophytic fungi

[0033] Take the bark of Juglans catalpa and scrape off the floating skin, rinse it under tap water for 20 minutes, place it on a sterile ultra-clean workbench, rinse it with sterile water 3 to 4 times, soak it with 75% alcohol by volume for 3 to 5 minutes, and then Sterilize with 1% NaClO solution for 20 minutes, then use sterile water to wash repeatedly 4 to 5 times, blot the sterile water on the bark with sterile paper, cut off the black surface, and cut out 0.5cm×0.5 The small pieces of cm are placed on the water agar solid plate medium and cultured in a 25°C incubator.

[0034] Take the newly grown hyphae at the incision, and transfer them to fresh PDA medium for culture in time. After the colonies grow, pick the colony edges on each plate according to the differences in colony shape, color and growth time. The hyphae at the place were inoculated on a new PDA plate for isolation and culture. The tip of the cultured hyphae is cut off...

Embodiment 2

[0036] Identification of Endophytic Fungi in Catalpa

[0037] Colony morphology: On the solid medium, the colony produces white fluffy mycelium, which is distributed radially, and the hyphae are well developed, such as figure 1 shown. Uniform round fungus balls are formed in the liquid culture solution, the surface of the fungus balls is rough and filamentous, the texture is loose, and the fermentation broth is yellow.

[0038] Spore Morphology: The spores are green in naked eyes, and ellipsoid in microscopic observation, with smooth surface. The main shaft of the conidiophores is long and strong, the secondary branches are short, and the branches are altar-shaped, the stalks are often solitary, and the spores are born on the top of the secondary branches. Under the microscope, the main axis of the conidiophores is long, the secondary branches inserted are short, and the secondary branches are few, and the branches are altar-shaped and the stalk is often solitary; in additio...

Embodiment 3

[0043] Preparation of Fermentation Broth Inhibiting the Growth of Liver Cancer Cells

[0044] Cultivate the isolated Trichoderma longibrachiata CCTCC M 208102 on a PDA plate, then cut a small piece of the PDA medium with mycelium, put it on the CMX medium, and cultivate it at room temperature. After two days, spores can be seen grow out. Then pick the spores and transfer them to a Erlenmeyer flask equipped with 250ml of CM liquid medium, culture them on a shaking table at 26°C to 28°C at 180rpm, stop the fermentation after 7-8 days, and obtain the spores that can inhibit the growth of human liver cancer cell line HepG2. fermentation broth. The bacterial fluid was collected for preliminary analysis of antitumor activity.

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Abstract

The invention discloses a preparation method in the technical field of molecular biology, which comprises following steps: (1) separation of endophytic fungi: the endophytic fungi are separated and purified by adopting a hyphal tip cutting method; (2) induction of spore bearing: long-branch trichoderma with the preservation number of CCTCC M 208102 is inoculated onto a PDA culture medium for being cultivated, a block of the hyphe-bearing PDA culture medium is cut, placed on a CMX culture medium and cultivated at room temperature, and spores are generated after 1 to 3 days; and (3) fluid fermented cultivation: the spores are selected, transplanted into a CM-containing fluid culture medium and oscillating cultivated on a shaking table at the temperature of 26 to 28 DEG C and the speed of 180rpm, and fermentation is terminated after 7 to 8 days, thus obtaining a fermented fluid with the inhibitive action of liver cancer cell growth. The preparation method is suitable for being used in the industrially scaled fermented production of the fermented fluid, which has the inhibitive action of HepG2 growth of liver cancer cell strains as well as high efficiency and low toxicity.

Description

technical field [0001] The invention relates to a preparation method in the field of biotechnology, in particular to a preparation method of a fermentation liquid for inhibiting the growth of liver cancer cells. Background technique [0002] Liver cancer is a disease with a high mortality rate in the world. The general treatment method is surgical resection. However, in many cases, it occurs centrally, and early metastasis of cancer cells is very easy to occur. This condition limits the application of surgical resection. Therefore, adjuvant therapy with some drugs is needed, but the effect of existing drugs on systemic chemotherapy for liver cancer patients is poor, so finding new effective chemotherapy drugs is of great significance to improve the survival rate of patients with advanced and recurrent liver cancer. [0003] Plant endophytic fungi are a class of fungi that live in the stems and leaves of host plants and complete their life cycle. During the evolution process,...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12P1/02C12N1/14A61P35/00C12R1/885
Inventor 苗志奇李美芽唐克轩
Owner SHANGHAI JIAOTONG UNIV