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Application of enterovirus type-71 in preparing oncolytic medicine

A technology of enteroviruses and drugs, applied in the field of oncology and virology, to achieve the effects of tumor growth inhibition, volume reduction, and survival rate improvement

Pending Publication Date: 2020-05-08
WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there is no report on the oncolytic activity of enterovirus 71 on tumor cells. The present invention provides a more effective solution for anti-tumor drug regimens by studying the oncolytic activity of enterovirus 71

Method used

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  • Application of enterovirus type-71 in preparing oncolytic medicine
  • Application of enterovirus type-71 in preparing oncolytic medicine
  • Application of enterovirus type-71 in preparing oncolytic medicine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Oncolytic effect of enterovirus 71 on different monolayers of tumor cells:

[0034] Enterovirus 71 BrCr infection (MOI=20) monolayer human neuroblastoma cell line SK-N-SH (SK), microglioma cell line T98G, glioma cell line CCF-STTG1 (CCF ) and the rhabdomyosarcoma cell line RD, by lactate dehydrogenase (LDH) cytotoxicity assay to evaluate the cell lysis response caused by viral oncolysis. Viral oncolysis causes tumor cell apoptosis or necrosis, resulting in damage to the cell membrane structure, resulting in the release of LDH in the cytoplasm into the culture medium. Quantitative analysis of tumor cell lysis responses can be achieved by detecting the activity of LDH released from cells with ruptured plasma membranes into the culture medium. The calculation method is as follows: tumor cytotoxicity index (%)=(absorbance of virus-treated sample-absorbance of control well) / (absorbance of maximum cell enzyme activity-absorbance of control well)×100. We respectively detecte...

Embodiment 2

[0037] Oncolytic effects of different genotypes of enterovirus 71 on monolayer glioma cells:

[0038] 1) EV71 has the targeting ability to kill tumor cells

[0039] Enterovirus type 71 BrCr strain infected (MOI=1) human glioma cell line CCF-STTG1 and normal glial cells (cells without any treatment as control, Mock group), and the cytopathic changes were observed through an inverted microscope , LDH assay to detect cell lysis reaction, one-step growth curve to detect virus amplification.

[0040] The result is as figure 2 As shown in middle A, EV71 infection of glioma cells can cause significant cytopathic reactions. In contrast, EV71 infection of normal glial cells only causes local cell rounding and shrinkage. Such as figure 2 As shown in middle B, after 48 hours of EV71 infection, the cytotoxicity indexes of glioma cells and normal glial cells were 60.2% and 8.2%, respectively, and the difference was extremely significant (P﹤0.001); after 96 hours of EV71 infection, the...

Embodiment 3

[0049] Oncolytic effect of enterovirus 71 in three-dimensional glioma cell spheres:

[0050] Add 200 microliters of tumor cell suspension (0.5×10 4 cells / ml), 37°C and 5% CO 2 After culturing for 4 days under certain conditions, tumor cell pellets with a diameter of about 500 microns were obtained. Enterovirus 71 infects tumor cell pellets, observes morphological changes with an inverted microscope, and detects cell viability with CCK-8 assay. The calculation method of cell viability is as follows: relative cell viability of tumor cell spheroids (%)=(absorbance of virus-treated tumor cell spheroid samples-absorbance of control wells) / (absorbance of untreated tumor cell spheroid samples-absorbance of control wells)×100. We detected the relative cell viability of the tumor cell spheroids after 4d and 7d of virus infection respectively.

[0051] Such as Figure 4 As shown in middle A, after EV71 infection, the integrity of the tumor cell spheres can be destroyed, making the e...

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Abstract

The invention relates to the field of oncology and virology and in particular discloses application of an enterovirus type-71 in preparing an oncolytic medicine. The invention finds for the first timethat the enterovirus type-71 has an oncolytic effect, in addition, has effects on multiple tumors such as glioma, neuroblastoma, rhabdomyosarcoma, liver cancer, lung cancer, colon cancer, breast cancer and ovarian cancer, has a specific killing function on tumor cells, but has no conspicuous killing function on normal neurogliocyte, and can be adopted to prepare a postoperative neoplasm recurrence medicine or a medicine for treating cancers.

Description

technical field [0001] The present invention relates to the fields of oncology and virology. Specifically, it relates to the application of enterovirus 71 in the preparation of oncolytic drugs. Background technique [0002] Human enteroviruses belong to the genus Enterovirus of the order Picornavirales and the family Picornaviridae. In the latest virus classification report released by the International Committee on Taxonomy of Viruses (ICTV), Enteroviruses are divided into 12 species. Enterovirus C is the type species of the Enterovirus genus, including polioviruses (PV-1, PV-2 and PV-3). [0003] Enterovirus 71 (EV71) belongs to the Enterovirus D species. The genome of EV71 is a single-stranded positive-strand RNA molecule with a length of about 7400nt, including the 5' non-coding region (5'UTR), the 3' non-coding region (3'UTR) and the viral polyprotein between the two single open reading frame. After EV71 infects host cells, it inhibits the synthesis of cellular pro...

Claims

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Application Information

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IPC IPC(8): A61K35/765A61P35/00
CPCA61K35/765A61P35/00
Inventor 崔宗强张晓玮李炜王汉中
Owner WUHAN INST OF VIROLOGY CHINESE ACADEMY OF SCI
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