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Fusobacterium nucleatum obtained from large intestine cancer tumor tissue and application of fusobacterium nucleatum obtained from large intestine cancer tumor tissue

A Fusobacterium nucleatum and tumor tissue technology, applied in tissue culture, tumor/cancer cells, bacteria, etc.

Active Publication Date: 2020-03-24
SHANGHAI TENTH PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Diverse Fusobacterium nucleatum strains currently lacking for colorectal cancer research

Method used

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  • Fusobacterium nucleatum obtained from large intestine cancer tumor tissue and application of fusobacterium nucleatum obtained from large intestine cancer tumor tissue
  • Fusobacterium nucleatum obtained from large intestine cancer tumor tissue and application of fusobacterium nucleatum obtained from large intestine cancer tumor tissue

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Experimental program
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Effect test

Embodiment 1

[0028] THCT14E2 was co-cultured with the colorectal cancer HCT116 cell line, and the specific steps were as follows:

[0029] The cultured HCT116 cells were taken, digested with trypsin to prepare HCT116 cell suspension, counted and inoculated into 24-well plates (1×10 5 cells / well), cultured for 12-24 hours until the cells adhered. After adhering to the wall, wash with PBS and replace the medium without anti-antibody. In the experimental group, freshly prepared THCT14E2 bacteria were added and suspended in PBS to make 1×10 per well 8 CFU; the control group was added with an equal volume of PBS. Three replicate holes were made for each group. cultured under normal conditions. The cell proliferation activity was detected by CCK8 method at 6 hours, 24 hours, 48 ​​hours, and 72 hours of culture respectively. The bacterial suspension was replaced every 24 hours.

[0030] Depend on figure 1 It can be seen that the proliferation activity of HCT116 cells at 72 hours was signif...

Embodiment 2

[0032] THCT14E2 was co-cultured with colorectal cancer LoVo cell line, the specific operation steps are as follows:

[0033] The cultured LoVo cells were taken, digested with trypsin to prepare LoVo cell suspension, counted and inoculated into 24-well plates (1×10 5 cells / well), cultured for 12-24 hours until the cells adhered. After adhering to the wall, wash with PBS and replace the medium without anti-antibody. Freshly prepared THCT14E2 bacteria were suspended in PBS, and the bacterial suspension was added to the experimental group (1×10 8 CFU / well); the control group was added with an equal volume of PBS. Three replicate holes were made for each group. cultured under normal conditions. The cell proliferation activity was detected by CCK8 method at 6 hours, 24 hours, 48 ​​hours, and 72 hours of culture respectively. The bacterial suspension was replaced every 24 hours.

[0034] Depend on figure 2 It can be seen that the proliferation activity of LoVo cells at 72 hou...

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Abstract

The invention provides fusobacterium nucleatum THCT14E2 which is obtained from large intestine cancer tumor tissue and does not belong to known fusobacterium nucleatum subsp.vincentii. The 16SrRNA gene sequence of the strain is as shown in SEQID NO:1. The classification designation of the THCT14E2 is Fusobacterium nucleatum, the preservation number is CCTCC NO:M 2019361, the preservation date is May 17th, 2019, and the preservation unit is China Center for Type Culture Collection. The colorectal cancer cell line and THCT14E2 co-culture result shows that the THCT14E2 can significantly promote colorectal cancer cell proliferation. Therefore, the THCT14E2 can promote understanding of micro-ecology diversity of rectum cancer, and facilitate function research of the fusobacterium nucleatum fromthe colorectal cancer.

Description

technical field [0001] The invention relates to the field of Fusobacterium nucleatum, in particular to a strain of Fusobacterium nucleatum obtained from colorectal cancer tumor tissue and its application. Background technique [0002] Colorectal cancer is a cancer with a high incidence in the world, and the disease burden caused by it has always been a difficult problem that scientists strive to overcome. At present, China has become the country with the largest number of new cases of colorectal cancer each year. Previous epidemiological surveys and experimental studies have generally confirmed that only 5%-15% of the incidence of colorectal cancer is attributed to genetic factors, and the remaining 85% is mainly affected by the environment, such as changes in dietary structure and intestinal microecological imbalance. Researches on colorectal cancer and intestinal microecological disorders are emerging, among which Fusobacterium nucleatum in the intestine is closely relate...

Claims

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Application Information

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IPC IPC(8): C12N1/20C12N5/09C12R1/01
CPCC12N1/20C12N5/0693C12N2500/72C12N1/205C12R2001/01
Inventor 秦环龙蔚青毕德玺
Owner SHANGHAI TENTH PEOPLES HOSPITAL
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