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Pediococcus acidilactici with anti-cervical cancer effect and application

A technology for Pediococcus lactis and cervical cancer, applied in the field of microorganisms, can solve problems such as different mechanisms

Active Publication Date: 2020-10-30
TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, their mechanisms are different, and the Pediococcus lactobacillus that can inhibit the expression of E6 and E7 and have anti-cervical cancer effects has not been reported yet.

Method used

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  • Pediococcus acidilactici with anti-cervical cancer effect and application
  • Pediococcus acidilactici with anti-cervical cancer effect and application
  • Pediococcus acidilactici with anti-cervical cancer effect and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] Embodiment 1: Screening and identification of strains

[0036] (1) Isolation and purification of strains

[0037] Dilute Maotai-flavored liquor fermented grains with sterile PBS buffer, spread on MRS plate medium, culture at 37°C for 48 hours under anaerobic conditions, and pick well-separated single colonies for streaking. A single colony was picked by streaking multiple times, and the purified strain was stained with Gram staining, and the morphology and distribution of the bacteria were observed under a microscope. This strain is a Gram-positive round coccus, see figure 1 .

[0038] The bacterial solution was diluted 10 times with sterile saline to 10 -6 , take 10 -4 、10 -5 、10 -6 100 μl of the three gradient dilutions were spread on the MRS solid plate, cultured at 37°C for 48 hours, and colonies of different shapes were selected for streak separation until pure single colonies with the same colony shape were obtained, and single colonies were picked and inocu...

Embodiment 2

[0043] Example 2: Effect of Pediococcus lactis HAO2018 cell lysate on the proliferation of cervical cancer cells

[0044] (1) Collect cervical cancer cells in the logarithmic growth phase in an ultra-clean bench, adjust the density of the cells with DMEM high-glucose medium, pipette 100 μL of cell suspension into a 96-well plate, and put it in 5% CO 2 , 37 ℃ cell culture incubator for 24h.

[0045](2) Cultivate overnight until the cells are completely adhered to the wall; after the cells are completely adhered to the wall, except the control group, the other groups are added with equal volumes and different concentration gradient samples of the supernatant of the lysate of Pediococcus lactis HAO2018, and 3 to 6 complexes are set. Wells were placed in an incubator for 24, 48, and 72 hours of culture respectively.

[0046] (3) After culturing for a corresponding period of time, discard the medium and wash it with PBS for 2 to 3 times, add 10 μL MTT to each well in the dark, inc...

Embodiment 3

[0048] Example 3: Effect of Pediococcus lactis HAO2018 cell lysate on apoptosis of cervical cancer cells

[0049] Inoculate 1×10 5 HeLa cells were cultured overnight in a 96-well plate. Except for the control wells, 100uL of Pediococcus lactis HAO2018 lysate supernatant was added to each well at a concentration of 10mg / mL to treat the cells, and the cells were harvested after 48h with 4% polymer Fix with formaldehyde for 30 min, incubate with TritonX-100 cell permeation agent at room temperature for 15 min, add TUNEL detection solution, incubate at 37°C in the dark for 60 min, add 100 μL of DAPI (1:100) solution to each well, incubate at room temperature in the dark for 5 min, and then Observe and take pictures under a focusing microscope.

[0050] The result is as image 3 As shown, it can be seen from the results of TUNEL staining that the supernatant of the lysate of Pediococcus lactis HAO2018 can cause DNA fragmentation of cervical cancer HeLa cells, and the number of po...

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Abstract

The invention relates to pediococcus acidilactici with an anti-cervical cancer effect. The name of the pediococcus acidilactici is HAO2018; the classification name is Pediococcus acidilactici; the preservation number is CGMCC No.16658, the preservation date is October 30, No. 3, Yard 1, West Beichen Road, Chaoyang District, Beijing, and the preservation unit is China General Microbiological Culture Collection Center. The pediococcus acidilactici can inhibit proliferation of cervical cancer HeLa cells, cause apoptosis of the HeLa cells, inhibit expression of human papilloma virus HPVE6 and E7 proteins and inhibit acetylation modification of histone in Hela cells. The Pediococcus acidilactici has good cervical cancer cell inhibition performance, is used for preparing a cervical cancer inhibition fermentation product and has a very wide application prospect.

Description

technical field [0001] The invention belongs to the technical field of microbes, in particular to Pediococcus lactis with anti-cervical cancer effect and its application. Background technique [0002] Cervical cancer is the second most common female malignancy worldwide, second only to breast cancer in incidence. Currently recognized as the main pathogenic factor of cervical cancer is the infection of high-risk human papillomavirus (HPV). Oncoproteins E6 and E7 encoded by high-risk HPV are the main carcinogenic factors, and their expression is affected by histone modification. The vaginal flora of healthy women is mainly composed of Firmicutes, including Gram-positive bacteria and Gram-negative bacteria. Under normal conditions, the vaginal flora is a dynamic balance system with lactic acid bacteria as the main dominant flora and constitutes the first line of defense against pathogens. Compared with the complex gastrointestinal flora, the composition of vaginal flora is r...

Claims

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Application Information

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IPC IPC(8): C12N1/20A61K35/741A61P35/00C12R1/01
CPCA61K35/741A61P35/00C12R2001/01C12N1/205
Inventor 何红鹏张同存王楠郝云鹏扈士月张潇李中媛宋亚囝
Owner TIANJIN UNIVERSITY OF SCIENCE AND TECHNOLOGY
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