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Fusobacterium nucleatum polymorphic subspecies isolate and application thereof

A technology of fusobacterium nucleatum and isolates, which is applied in the direction of bacteria, analytical materials, biological material analysis, etc., can solve the problem of not being able to represent the actual characteristics of fusobacterium nucleatum, and achieve the effect of promoting proliferation

Active Publication Date: 2020-04-14
SHANGHAI TENTH PEOPLES HOSPITAL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Therefore, they may not represent the actual characteristics of F. nucleatum in the gut
At present, Fusobacterium nucleatum, especially the polymorphic subspecies, derived from colorectal cancer tumor tissue has rarely been reported

Method used

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  • Fusobacterium nucleatum polymorphic subspecies isolate and application thereof
  • Fusobacterium nucleatum polymorphic subspecies isolate and application thereof

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Experimental program
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Effect test

Embodiment 1

[0026] THCT15E1 was co-cultured with colorectal cancer HCT116 cell line, and the specific operation steps were as follows:

[0027] The cultured HCT116 cell suspension was prepared by trypsinization, counted and inoculated into a 24-well plate (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 THCT15E1 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.

[0028] Depend on figure 1 It can be seen that the proliferation activity of HCT116 cells at 72 hours was signifi...

Embodiment 2

[0030] THCT15E1 was co-cultured with colorectal cancer LoVo cell line, the specific operation steps are as follows:

[0031] The cultured LoVo cell suspension was prepared by trypsinization, 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 THCT15E1 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.

[0032] Depend on figure 2 It can be seen that the proliferation activity of LoVo cells at 72 hours was significantly high...

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Abstract

The invention provides a fusobacterium nucleatum polymorphic subspecies isolate THCT15E1, a classification name is Fusobacterium nuucleates subsp.nucleatum THCT15E1, a preservation number of the strain is CCTCC NO: M 2019362, a preservation date is May 17, 2019, a preservation unit is China Center for Type Culture Collection, and the 16SrRNA gene sequence of the strain is shown as SEQ ID NO: 1. The result of co-culture of the colorectal cancer cell line and THCT15E1 shows that THCT15E1 can significantly promote colorectal cancer cell proliferation; therefore, the THCT15E1 can provide diversified experimental conditions for in-vitro or in-vivo intestinal environment simulation for colorectal cancer research, and can also construct a colorectal cancer disease model to screen drugs for treating colorectal cancer.

Description

technical field [0001] The invention relates to the field of Fusobacterium nucleatum subsp. polymorpha, in particular to an isolate of Fusobacterium nucleatum subsp. polymorpha and application thereof. Background technique [0002] Colorectal cancer is one of the most common tumors, and its mortality rate ranks third among all tumors. The burden of disease caused by it has always been a difficult problem that scientists strive to overcome. With the development of omics technologies such as sequencing, the role of microorganisms in the development of colorectal tumors has attracted increasing attention, and the close relationship between Fusobacterium nucleatum and the development of colorectal tumors has been verified in many ways. [0003] Fusobacterium nucleatum is a Gram-negative anaerobic bacterium that routinely colonizes the oral cavity and is normally rare elsewhere in the body. Fusobacterium nucleatum was first identified in the high bacterial load of periodontitis...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N1/20C12N5/09C12Q1/02C12R1/01
CPCC12N5/0679C12N5/0693C12N2502/70C12N2503/02C12Q1/025C12N1/205C12R2001/01G01N33/5011G01N2333/195G01N2500/10
Inventor 秦环龙蔚青毕德玺
Owner SHANGHAI TENTH PEOPLES HOSPITAL
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