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Lactococcus lactis subsp. lactis for regulating signal path of colon tumor and application thereof

A technology of Lactococcus lactis and lactic acid subspecies, applied in the field of microorganisms, can solve the problems of not being able to be used in food, and the regulation of tumor-related signaling pathways is not clear, so as to reduce inflammation, have good gastric acid resistance, bile salt resistance, and good tolerance effect of ability

Active Publication Date: 2018-01-19
JIANGNAN UNIV YANGZHOU FOOD BIOTECH INST +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] CN104523761A discloses a genetically engineered strain of Lactococcus lactis capable of inducibly expressing interleukin 12, which can inhibit the growth of intestinal tumors, but this bacterium is not obtained from nature and can only be used for medicine and not for food
The above three patented strains are clear and have clear functions, but the animal model involved uses colon cancer cell lines subcutaneously bearing tumors, which is quite different from the tumor environment in the intestine, and the role in the regulation of tumor-related signaling pathways is not clear

Method used

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  • Lactococcus lactis subsp. lactis for regulating signal path of colon tumor and application thereof
  • Lactococcus lactis subsp. lactis for regulating signal path of colon tumor and application thereof
  • Lactococcus lactis subsp. lactis for regulating signal path of colon tumor and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1: Lactococcus lactis subspecies VM5 has good tolerance to simulated gastrointestinal fluid

[0066] Streak-inoculate the cryopreserved Lactococcus lactis subsp. lactic acid VM5 in the solid medium of MRS medium, culture it aerobically at 37°C for 48 hours, and subculture it in MRS medium for 2 to 3 times, then take Lactococcus lactis Lactic acid subspecies VM5 culture fluid was collected by centrifugation at 8000×g for 5 min, resuspended in (1:1) artificial simulated gastric juice of pH 2.5 (MRS medium containing 1% pepsin, pH=2.5), and then mixed in Anaerobic culture was carried out at 37°C. Samples were taken at the beginning (0h), 1h, 2h and 3h respectively, and MRS agar medium was poured for culture to count plate colonies, determine the number of viable bacteria and calculate their survival rate. The survival rate is the ratio of the number of viable bacteria in the culture medium to the number of viable bacteria at 0h, expressed in %.

[0067] Take the c...

Embodiment 2

[0072] Example 2: Regulation of Lactococcus lactis subsp. lactic acid VM5 on Notch1, VEGFR2 and colon cancer-related signaling pathways in HT-29 cells

[0073] Use RPMI1640 cell culture medium without antibiotics to resuspend the bacteria sludge of Lactococcus lactis subsp. lactis VM5 (or control Lactococcus lactis subsp. lactis LM35, LGG, Escherichia coli E.coli) washed twice with PBS, and adjust the bacteria Bulk density to about 2×10 8 CFU / mL; add VM5 (or LM35, LGG, E.coli) suspension into the 6-well plate cultured with HT-29 according to the amount of 2mL bacterial suspension per well (cell fusion degree reaches 95%), blank group Add 2 mL of antibiotic-free cell culture medium and place at 37 °C with 5% CO 2 Incubate in a cell culture incubator for 2 hours; wash the cells 3 times with PBS, add 1 mL TRIzol to each well (6-well plate), let stand at room temperature for 5 minutes, repeatedly pipette with an enzyme-free pipette tip, and then transfer to 1.5 mL EP without enzy...

Embodiment 3

[0078] Example 3: Lactococcus lactis subspecies VM5 has no acute toxic side effects on mice

[0079] Lactococcus lactis subspecies VM5 was resuspended in 2% (w / v) sucrose solution, and the cell density was 4.0×10 9 CFU / mL. Take 10 healthy male BALB / c mice with a body weight of about 25 g, give the suspension of this concentration orally once a day, observe for a week, and record the death and body weight.

[0080] The results of these tests are listed in Table 4. These results indicated that feeding concentrations of 4.0×10 9 CFU / mL of Lactococcus lactis subsp. lactic acid VM5 had no significant impact on the mice, no significant change in body weight, and no death. The appearance of the mice had no obvious pathological symptoms.

[0081] Table 4 Changes in body weight and death conditions of mice

[0082] time (days)

[0083] Note: -: no death of mice

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Abstract

The invention discloses a lactococcus lactis subsp. lactis for regulating a signal path of colon tumor and an application thereof and belongs to the technical field of microorganisms. The lactococcuslactis subsp. lactis VM5 provided by the invention is preserved with the preservation number of GDMCC No.60256, is capable of enduring gastric acid and cholate, is capable of obviously relieving the inflammatory response of the colorectal part of a colorectal cancer model rat and is capable of reducing the tumor quantity on the colorectal parts of the model rat in the manner of regulating the expression of Notch1 signal path and VEGFR2 molecule in the colorectal tissue. Besides, the lactococcus lactis subsp. lactis VM5 is capable of improving the short chain fatty acid level in intestinal flora structures and intestinal tracts. The lactococcus lactis subsp. lactis VM5 is used for preparing the fermented food for restraining the happening of colorectal cancer and has a wide application prospect.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a Lactococcus lactis subsp. lactis capable of significantly inhibiting the occurrence of colorectal cancer by regulating Notch1 and VEGFR2 signaling pathways and its application. Background technique [0002] Colorectal cancer is a malignant gastrointestinal tumor. As the morbidity and mortality of colorectal cancer continue to rise in my country, its threat to the health of our people is gradually deepening. At present, the mainstream methods for the clinical treatment of colorectal cancer all have relatively large side effects, which greatly affect the quality of life of patients. As a safe edible food, probiotics have been gradually confirmed by researches to alleviate and treat various diseases, such as promoting digestion and absorption, enhancing immunity, preventing reproductive system infections, alleviating allergic reactions, preventing and inhibiting Tumors, p...

Claims

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Application Information

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IPC IPC(8): C12N1/20A23C9/12A23L11/00A23C11/10A23L19/20A23L33/00A23L33/135A61K35/744A61P35/00C12R1/01A23L11/50A23L11/60
Inventor 陈卫王刚赵建新张灏
Owner JIANGNAN UNIV YANGZHOU FOOD BIOTECH INST
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