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Use of Lactoferrin in Prevention and Treatment of Tumor Metastasis

A technology of lactoferrin and human lactoferrin is applied in the field of preventing and inhibiting tumor metastasis, which can solve the problems of lack of methods and drugs for tumor metastasis.

Active Publication Date: 2015-09-23
SHANGHAI TRANSGENIC RES CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] In recent years, the research on the mechanism of tumor metastasis has made progress and breakthroughs to a certain extent, but there is still a lack of satisfactory methods and drugs for effectively inhibiting tumor metastasis (especially lung metastasis) in this field.

Method used

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  • Use of Lactoferrin in Prevention and Treatment of Tumor Metastasis
  • Use of Lactoferrin in Prevention and Treatment of Tumor Metastasis
  • Use of Lactoferrin in Prevention and Treatment of Tumor Metastasis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0075] The preparation of embodiment 1 recombinant human lactoferrin

[0076] 1 Materials and methods

[0077] 1.1 Reagents and raw materials

[0078] buffer A (400mM NaCl, 20mM PB, pH7.5), buffer B (1000mM NaCl, 20mM PB, pH7.5); physiological saline.

[0079] The transgenic goat milk of recombinant human lactoferrin (purchased from Shanghai Transgenic Research Center) was determined to contain about 40-66 g of human lactoferrin and a small amount of about 0.05-0.12 g of goat lactoferrin per liter of goat milk.

[0080] 1.2 Method

[0081] 1.2.1 Degreasing:

[0082] Take 20 mL of transgenic goat milk highly expressing recombinant human lactoferrin, and degrease the goat milk as follows: centrifuge at 10,000 rpm for 30 min, and filter with 4 layers of gauze to obtain the filtrate.

[0083] 1.2.2 Cheese off:

[0084] Mix the filtrate with 20mL 800mM NaCl (40mM PB, pH7.5), adjust the pH to 4.6 with concentrated hydrochloric acid; after incubating at 40°C for 30min, centrifug...

Embodiment 2

[0097] Example 2 In vitro growth inhibition test and metastasis inhibition test of recombinant human lactoferrin on non-small cell lung cancer A549 cell line

[0098] 1 Materials and methods

[0099] 1.1 Reagents and materials

[0100] Transwell chamber (pore size 8 μm), DMEM basal medium, DMEM complete medium, DMEM medium (containing 1% fetal bovine serum), DMEM medium (containing 20% ​​fetal bovine serum), trypsin, penicillin and streptomycin, 0.1 % crystal violet staining solution, MTT.

[0101] Non-small cell lung cancer A549 cell line.

[0102] 1.2 Method

[0103] 1.2.1 In vitro growth inhibition test of rhLF on non-small cell lung cancer A549 (MTT method)

[0104] Collect logarithmic phase A549 cells, adjust the concentration of cell suspension to 10 4 cells / mL, add 100 μL per well to make the cell density to be tested as 1000 wells. 5%CO 2 , incubate at 37°C until the cells adhere to the wall, add concentration gradient rhLF, 100 μL per well, the final drug conce...

Embodiment 3

[0114] Example 3 Recombinant human lactoferrin inhibits the growth of mouse melanoma lung metastases

[0115] In order to further study the activity of lactoferrin in inhibiting tumor metastasis in animals, in this example, the mouse melanoma B16F10 lung metastasis model was used to evaluate the activity of lactoferrin in inhibiting tumor metastasis, colonization and growth in vivo.

[0116] 1 Materials and methods

[0117] 1.1 Experimental animals

[0118] Animal source: C57BL / 6 mice, provided by the Experimental Animal Research Center of the Chinese Academy of Military Medical Sciences (Experimental Animal Production License: SCXK (Army))

[0119] Animal age: 6-8 weeks; animal weight: 18-22g; animal sex: male.

[0120] 1.2 Test drugs

[0121] 1) Human lactoferrin

[0122] Purified from transhuman lactoferrin gene goat milk, preparation method: prepare the required concentration with physiological saline before each administration. The purification method is the same as ...

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Abstract

The invention provides the usage of lactoferrin in prevention and treatment of tumor metastasis. Particularly, the invention provides the usage of the lactoferrin in preparation of a medicine for preventing or inhibiting tumor metastasis or tumor cell migration. The pharmacology experiment in the invention proves that the human lactoferrin has good prevention and treatment effects for the tumor metastasis, so that metastasis, colonization and growth of malignant tumors such as mouse melanoma and the like can be obviously inhibited.

Description

technical field [0001] The invention relates to the field of medicine, especially the fields of biological pharmacy and tumor treatment. More specifically, the present invention relates to a method for preventing and inhibiting tumor metastasis by administering human lactoferrin or a composition combined with other anticancer drugs. Background technique [0002] Lactoferrin (Lactoferrin, LF) is a multifunctional basic glycoprotein with a molecular weight of about 80KD, which mainly exists in mammalian exocrine fluids (such as: milk, tears, sweat, saliva, semen, etc.) and neutrophils . LF is abundant in human milk, the content in colostrum can reach 7g / L, and the content in mature milk decreases and tends to be balanced, usually at the level of 1g / L. [0003] Since LF was first isolated from cow's milk in 1939, its biological activities such as promoting the absorption and delivery of iron in the body, antibacterial, antiviral infection, and regulating the body's immunity h...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K38/40A23L1/29A61P35/04A23L33/00
Inventor 成国祥孙伟刘思国陈建泉
Owner SHANGHAI TRANSGENIC RES CENT
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