A kind of Lactobacillus plantarum capable of inhibiting the occurrence of colorectal cancer and its use

A technology of Lactobacillus plantarum and colon cancer, applied in the field of microorganisms, can solve the problems of inability to be used in food, unclear regulation of tumor-related signaling pathways, etc., achieve the level of relieving inflammation, good resistance to gastric acid and bile salts, and reduce inflammation. Effect

Active Publication Date: 2020-05-22
JIANGNAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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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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  • A kind of Lactobacillus plantarum capable of inhibiting the occurrence of colorectal cancer and its use
  • A kind of Lactobacillus plantarum capable of inhibiting the occurrence of colorectal cancer and its use
  • A kind of Lactobacillus plantarum capable of inhibiting the occurrence of colorectal cancer and its use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0065] Example 1: Lactobacillus plantarum CCFM164 has good tolerance to simulated gastrointestinal fluid

[0066] Streak inoculation of cryopreserved Lactobacillus plantarum CCFM164 in MRS solid medium, aerobic static culture at 37°C for 24 hours, and subculture by MRS medium for 2 to 3 times, then take Lactobacillus plantarum CCFM164 culture medium, The bacteria were 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 cultured aerobically at 37°C. Samples were taken at the beginning (0h), 1h, 2h and 3h, and MRS agar medium was used to pour culture for plate colony counting, and the number of viable bacteria was determined and the survival rate was calculated. 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 culture solution of Lactobacillus plantarum CC...

Embodiment 2

[0072] Example 2: Regulation of Lactobacillus plantarum CCFM164 on Notch1, Notch2, VEGFR2 and colon cancer-related signaling pathways in HT-29 cells

[0073] Use antibiotic-free RPMI1640 cell culture medium to resuspend Lactobacillus plantarum CCFM164 (or control Lactobacillus plantarum LP60, LGG, Escherichia coli E.coli) sludge washed twice with PBS, and adjust the cell density to about 2× 10 8 CFU / mL; CCFM164 (or LP60, LGG, E.coli) suspension was added to the 6-well plate cultured with HT-29 according to the amount of 2 mL of bacterial suspension per well (the degree of cell fusion reached 95%), and the 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 enzyme-free According to the in...

Embodiment 3

[0078] Example 3: Lactobacillus plantarum CCFM164 has no acute toxic side effects on mice

[0079] Plantarum CCFM164 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 Lactobacillus plantarum CCFM164 had no significant effect on 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) 1 2 3 4 5 6 7 weight(g) 25.5±1.6 25.5±1.7 25.6±1.5 25.6±1.6 25.7±1.8 25.6±1.9 25.7±1.9 Circumstances of death - - - - - - -

[0083] Note: -: no...

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Abstract

The invention discloses a plant lactobacillus (Lactobacillus plantarum) capable of significantly inhibiting the occurrence of colorectal cancer and an application thereof, belonging to the technical field of microbes. The Lactobacillus plantarum CCFM164 provided by the present invention, whose preservation number is CGMCC No.14520, can tolerate gastric acid and bile salts, and can significantly reduce the inflammatory response in the colorectal part of colorectal cancer model mice, by regulating Notch1 in colorectal tissue , Notch2 signaling pathway and the expression of VEGFR2 molecules, reducing the number of tumors in the colon and rectum of model mice. In addition, Lactobacillus plantarum CCFM164 can also improve the structure of intestinal flora and the level of short-chain fatty acids in the intestine. The Lactobacillus plantarum CCFM164 is used to prepare fermented food that inhibits the occurrence of colorectal cancer, and has very broad application prospects.

Description

technical field [0001] The invention relates to the technical field of microorganisms, in particular to a lactobacillus plantarum capable of significantly inhibiting the occurrence of colorectal cancer by regulating Notch1, Notch2 and VEGFR2 signaling pathways and its use. 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, prevention...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A23L33/135A23L11/00A23L19/00A23C9/123A23C11/10A23C13/16A23C19/032C12R1/25A23L11/50A23L11/60
CPCA23C13/16A23L19/00A23L33/135C12N1/20C12R2001/25C12N1/205A61P1/14A23L27/50A23L11/50A23C9/1234A23C19/062A23C19/0323A23L19/20A61K9/0019A61K35/747A23L11/65Y02A50/30A23V2400/169A23L33/40A23V2002/00A61K9/0056
Inventor 陈卫王刚王尔胤赵建新张灏
Owner JIANGNAN UNIV
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