Application of ORFV in preparation of anti-colorectal cancer cell drugs

A technology for sheep infectious pustules and colorectal cancer, applied in the biological field, can solve the problems of targeted antibody drug resistance, high price, and high cost, and achieve good curative effect, low cost, and low toxicity

Inactive Publication Date: 2018-05-25
FOSHAN UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, targeted antibodies also have side effects such as drug resistance, hi

Method used

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  • Application of ORFV in preparation of anti-colorectal cancer cell drugs
  • Application of ORFV in preparation of anti-colorectal cancer cell drugs
  • Application of ORFV in preparation of anti-colorectal cancer cell drugs

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0035] ORFV virus effectively inhibits colorectal cancer tumor growth

[0036] It can replicate and proliferate in 6 kinds of colorectal cancer cell lines, and at the same time reduce the survival rate of tumor cells, and inhibit the growth and metastasis of 6 kinds of colorectal cancer cell lines, 6 kinds of human colon cancer cells Caco-2, HCT116, LoVo , RKO, SW480 and SW1116 were purchased from Shanghai Cell Bank (Shanghai, China). Mouse colon cancer cell CT26 is derived from BALB / c mice and kept in the laboratory. All colon cancer cells were cultured in DMEM medium containing 10% bovine serum.

[0037] The CCK-8 method was used to detect the tumor cells inoculated in a 96-well plate at a density of 103 per well, and the tumor cells were stimulated with ORFV with different virus titers (MOI=0, 1, 5, 10). Cells were harvested at 0, 24, 48 and 72 hours. Remove cell supernatant at 37 °C, 5% CO 2 Incubate with CCK-8 solution (10% CCK-8 diluted DMEM medium containing 10% bov...

Embodiment 2

[0041] ORFV virus causes tumor cell death in colorectal cancer

[0042] The apoptosis of tumor cells induced by ORFV virus was detected by flow cytometry. Tumor cells were inoculated in a 96-well plate at a density of 106 per well, and ORFV or PBS with virus titer (MOI=10) was added to culture for 24 hours, and the tumor cells were collected at 800RCF for 5 minutes. Adherent cells were digested with 0.25% trypsin-EDTA (Solabio, Beijing, China), washed twice with PBS, and collected in 1.5ml clean EP tubes. Cell apoptosis was detected by flow cytometry (BD LSR Fortessa).

Embodiment 3

[0044] ORFV inhibits the growth of subcutaneous tumor tissue in mice

[0045] CT26 (50 μL 0.9% NaCl / 5 ⅹ 105 ) cells were inoculated subcutaneously in Balb / c mice to establish a tumor model. When the tumor reached 200-250 mm3, the mice were randomly divided into two groups. One group was treated with intratumoral injection of 50 μl PBS, and the other group was treated with intratumoral injection of ORFV (108TCID50, 50 μL). Inject every 3 days. After 12 days, the mice were sacrificed, and the mouse tumors and spleens were collected. In the metastasis model, Balb / c mice were subcutaneously inoculated with CT26 (50 μL 0.9% NaCl / 5ⅹ10 5 )cell. After 24 hours, the mice were randomly divided into two groups. One group of mice was injected with ORFV (108 PFU) into the tail vein on day 1, 3 and 8 respectively, and the other group was injected with PBS intravenously. The mice were sacrificed after 11 days. Collect mouse spleen and serum.

[0046] The collected CT26 tumor and spleen ...

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Abstract

The invention discloses application of ORFV in preparation of anti-colorectal cancer cell drugs and application of the ORFV in effective inhibition of colorectal cancer cell growth. The gene-modifiedORFV is used as oncolytic virus, has strong inhibition effects on growth and metastasis of colorectal cancer cells, does not introgress into a host genome, and has the advantages of being good in safety to human and potential curative effect, low in cost and toxicity and capable of achieving repeated inoculation.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to the application of a sheep infectious impetigo virus in the preparation of anti-colorectal cancer cell medicines. Background technique [0002] Ovine infectious impetigo virus (commonly known as ovine oral ulcer virus, ORFV) belongs to Poxviridae, Parapoxvirus genus, and is the pathogen of ovine infectious impetigo (orfv). Goat contagious impetigo is a localized, highly contagious disease characterized by the formation of papules, pustules, ulcers and thick verrucous scabs on the skin and mucous membranes of affected sheep. Lambs are the most susceptible to disease, mostly in groups. Due to the strong resistance of the virus, once the flock is infected, it is not easy to remove, and it can continue to endanger the flock for many years, causing great economic losses to the sheep industry. ORFV virus particles are brick-shaped or oval-shaped, and their surface is arranged in a rope-l...

Claims

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Application Information

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IPC IPC(8): A61K35/768A61P35/00A61P35/04
CPCA61K35/768
Inventor 罗树红成敏郝文波陈达香龙銘坚王勇龚道元王瑞雪
Owner FOSHAN UNIVERSITY
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