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Application of novel oncolytic virus in preparation of medicine for treating colorectal cancer

An oncolytic virus, colorectal cancer technology, applied in the fields of biotechnology and biomedicine, can solve the problems of lack of specificity of infecting tumors, difficult to guarantee safety, unstable genetic characteristics of RNA oncolytic virus, etc., and achieve large clinical application. Promising, easy to handle, effect of significant oncolytic properties

Pending Publication Date: 2020-08-07
吴建国
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Through the above analysis, the existing problems and defects of the prior art are as follows: (1) The prior art lacks relevant technical basis for the therapeutic effect of EV71 on various cancer patients, including colorectal cancer patients
[0006] (2) Traditional surgical resection has limited therapeutic effect and is prone to recurrence
[0007] (3) Chemotherapy is limited by its toxicity to other normal tissues in the body, and radiotherapy radiation can also cause damage to normal tissues
[0008] (4) DNA oncolytic virus encodes an oncogene, which is easy to integrate into the host genome, and its safety is difficult to guarantee; and the viral genome is large, making it difficult to carry out genetic modification
However, the known RNA oncolytic viruses have unstable genetic characteristics and lack specificity for infecting tumors.

Method used

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  • Application of novel oncolytic virus in preparation of medicine for treating colorectal cancer
  • Application of novel oncolytic virus in preparation of medicine for treating colorectal cancer
  • Application of novel oncolytic virus in preparation of medicine for treating colorectal cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1: the killing effect detection of EV71 in vitro

[0061] The killing effect detection of EV71 on colorectal cancer cells, the steps are:

[0062] 1. Experimental materials:

[0063] 1.1 Cells and viruses:

[0064] DLD1, HCT116, LoVo, HT29, SW48 colorectal cancer cell lines were obtained from ATCC Company in the United States. Enterovirus 71 (EV71) strain (Xiangyang-Hubei-09) was derived from EV71-infected brain tissue in Xiangyang City, Hubei Province, China, and was previously isolated from the State Key Laboratory of Virology, Wuhan University (GenBank: JN230523.1). species expanded in RD cells.

[0065] 1.2 Reagents:

[0066] DMEM medium and fetal bovine serum (FBS) were purchased from GIBCO; CCK8 (Cell Counting Kit-8) detection kit was purchased from Sangon Bioengineering (Shanghai) Co., Ltd.

[0067] 1.3 Experimental equipment:

[0068] The microplate reader is a product of Thermo Company; the cell culture box is a product of Thermo Company.

[0...

Embodiment 2

[0082] Embodiment 2: In vitro EV71 detects the killing effect of normal intestinal cells

[0083] The present invention provides a kind of application of human enterovirus EV71 in human colorectal cancer, and its steps are:

[0084] 1. Experimental materials

[0085] 1.1 Cells and viruses

[0086] Human primary intestinal epithelial cells FHC and HCoEpic were purchased from ATCC, USA. EV71 (Human enterovirus71strain Xiangyang-Hubei-09) was isolated from the State Key Laboratory of Virology of Wuhan University (GenBank: JN230523.1), and the virus stocks were amplified in RD cells.

[0087] 1.2 Reagents

[0088] DMEM medium and FBS were purchased from GIBCO; CCK8 (Cell Counting Kit-8) detection kit was purchased from Sangon Bioengineering (Shanghai) Co., Ltd.

[0089] 1.3 Experimental Instruments

[0090] The microplate reader is a product of Thermo Company; the cell culture box is a product of Thermo Company.

[0091] 2. Experimental methods and results

[0092] 2.1 Dete...

Embodiment 3

[0101] Embodiment 3: the detection of the oncolytic effect of EV71 in vivo

[0102] The application of a kind of human enterovirus EV71 of the present invention in human colorectal cancer, its steps are:

[0103] 1. Experimental materials:

[0104] 1.1 Cells, viruses, nude mice:

[0105] HT29 cells were purchased from ATCC, USA. Nude mice (5 weeks old, male) were purchased from the Experimental Animal Center of Guangdong Province, China. EV71 (Human enterovirus 71strain Xiangyang-Hubei-09) was isolated from the State Key Laboratory of Virology of Wuhan University (GenBank: JN230523.1), and the virus stock was amplified in RD cells.

[0106] 1.2 Reagents:

[0107] DMEM medium, FBS and PBS buffer solution (pH=7.4) were purchased from GIBCO Company.

[0108] 1.3 Experimental equipment:

[0109] The cell incubator is a product of Thermo Company.

[0110] 2. Experimental methods and results:

[0111] 2.1 Construction of xenograft tumor transplantation model in nude mice:

...

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Abstract

The invention, which belongs to the technical field of biotechnology and biomedicine, discloses application of a novel oncolytic virus in preparation of a medicine for treating colorectal cancer. Thedosage of the novel oncolytic virus is (5-10)*10<5>PFU. The novel oncolytic virus is amplified in RD cells, and the virus is purified and concentrated by ultra-high-speed centrifugation. The enterovirus 71 can infect and lyse colorectal cancer cells in an in-vitro cell experiment, but has little influence on normal intestinal cells, which indicates that the EV71 has the advantages of specific infection and colorectal cancer cell lysis. The EV71 can inhibit tumor growth in nude mice in a xenograft tumor model, which indicates that the EV71 can inhibit tumor growth and delay tumor progression invivo. The EV71 is an RNA virus, does not encode cancer genes, is not integrated to a host genome, and is high in safety; and the virus cDNA is easy to operate, and virus genome modification is facilitated.

Description

technical field [0001] The invention belongs to the technical fields of biotechnology and biomedicine, and in particular relates to the application of a novel oncolytic virus in the preparation of a drug for treating colorectal cancer. Background technique [0002] At present, colorectal cancer (colorectal cancer) is a kind of intestinal malignant tumor originating from the colon or rectum, and its morbidity and mortality rate are among the highest in the world. The 5-year survival rate for colorectal cancer is about 90% when it is diagnosed early and 13% when it is delayed. Although colorectal cancer screening helps reduce the number of patients diagnosed with advanced cancer, most cases It can only be determined after the onset of symptoms. At present, in medicine, the treatment of cancer is mainly determined by the location of cancer, its stage, and the progress of cancer. Generally speaking, early cancer (solid tumor, pathological stage I-II, no or few lymph node metas...

Claims

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Application Information

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IPC IPC(8): A61K35/768A61P35/00C12Q1/02
CPCA61K35/768A61P35/00G01N33/5011G01N33/5014
Inventor 吴建国罗震阮志慧邬开朗谭秋萍
Owner 吴建国
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