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Antiviral effect and application of epigenetic modifying enzyme SETD2

A virus infection and virus infection-related technology, applied in the field of epigenetic modification enzyme SETD2, can solve the problem of unclear role of SETD2 protein, and achieve the effect of preventing and inhibiting virus infection, inhibiting virus infection and improving curative effect.

Active Publication Date: 2017-08-11
SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, so far, the role of SETD2 protein in immune response and antiviral infection is still unclear

Method used

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  • Antiviral effect and application of epigenetic modifying enzyme SETD2
  • Antiviral effect and application of epigenetic modifying enzyme SETD2
  • Antiviral effect and application of epigenetic modifying enzyme SETD2

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0107]Example 1: Interfering with the expression of SETD2 has a promoting effect on the replication of HBV in cells

[0108] HepG2 cells (purchased from ATCC) were cultured with DMEM medium, and the cells were transfected with HBV expression plasmids based on the framework of pHBV1.3, simulating the environment in which cells were infected with HBV. Then the cells were transfected with small interfering RNA against SETD2 (si-SETD2) and mock control (si-mimetic) (the transfection reagent INTERFERin was purchased from Polyplus).

[0109] The interfering RNA (Si-SETD2) against SETD2 and the mimic control (Si-mimetic) were purchased from Genephama Company, the sequence of Si-SETD2 is shown in SEQ ID NO: 5 and SEQ ID NO: 6, and the Si-mimetic The sequences are shown in SEQ ID NO: 7 and SEQ ID NO: 8. During synthesis, 2 dTs were added at the 3' to make the sequences more stable. The specific sequence is as follows:

[0110] Si-SETD2 sequence:

[0111] 5'-GGUGUAACUUAUGCAUUAATT-3' ...

Embodiment 2

[0120] Example 2: Overexpression of SETD2 inhibits HBsAg secretion in cells

[0121] Firstly, the cDNA of SETD2 (NM_014159.6) was divided into three segments and introduced into the eukaryotic expression vector pcDNA3.1 plasmid respectively, and the expression vectors of SETD2-F1, SETD2-F2 and SETD2-F3 were respectively constructed.

[0122] Wherein, the nucleotide sequence contained in SETD2-F1 is the 54th-2888th position of SEQ ID NO: 1, which encodes the 1st-945th position of human SETD2 protein SEQ ID NO: 2;

[0123] The nucleotide sequence contained in SETD2-F2 is SEQ ID NO: 1 2889-5267, which encodes human SETD2 protein SEQ ID NO: 2 946-1738, and this truncated body contains the SET domain;

[0124] The nucleotide sequence contained in SETD2-F3 is the 5268-7748 position of SEQ ID NO: 1, which encodes the 1739-2564 position of human SETD2 protein SEQ ID NO: 2, and this truncated body contains the WW domain.

[0125] SETD2-F1, SETD2-F2 and SETD2-F3 were respectively trans...

Embodiment 3

[0130] Example 3: Overexpression of SETD2 inhibits HBV infection in mouse liver

[0131] Construct SETD2-F2 expression vector (same as Example 2). 20ng SETD2-F2 was injected into mice (8-week-old male SDF-grade C57BL6 mice, purchased from Sipper BK) through high-pressure tail vein, and 10ng pHBV1.3 plasmid was injected into the tail vein at the same time to simulate HBV infection in mice. After 72 hours, IFNα (15 mg / kg) was injected intraperitoneally. After 24 hours, the mouse liver tissue was dissected, and the expression of HBV core antigen (HBcAg) on ​​the mouse liver was detected by immunofluorescence to indicate the HBV infection of the mouse.

[0132] The results of liver HBV-infected mice were as follows: Figure 4 shown.

[0133] The results showed that overexpression of SETD2-F2 in mice could inhibit HBV infection in mouse liver.

[0134] The results indicated that overexpression of SETD2 could inhibit virus infection in vivo.

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Abstract

The invention discloses an antiviral effect and an application of an epigenetic modifying enzyme SETD2. The invention relates to the fields of biotechnology and medicine, and in particular to an anti-virus infection effect and an application of an immuno-regulatory molecule SETD2. Specifically, the invention relates to the SETD2 or an SETD2 coding sequence and an application of an accelerant of the SETD2 in preparing a medicine or a kit for inhibiting virus infection, and further relates to applications of the material in resisting virus infection and treating chronic inflammatory diseases caused by the virus infection, an application method and the corresponding medicine, a pharmaceutical composition or the kit. The SETD2 provided by the invention can be used for preventing and inhibiting the virus infection and enhancing a curative effect of interferon in resisting the virus infection, and the SETD2 has a broad application prospect.

Description

technical field [0001] The present invention relates to the fields of biotechnology and medicine, in particular, an epigenetic modification enzyme SETD2, that is, protein 2 containing SET domain containing 2 (SET domain containing 2, SETD2) is useful in preventing or treating diseases or symptoms related to viral infection. Symptoms, effects, mechanisms of action, implementation methods and uses in controlling damage caused by virus infection. Background technique [0002] Infection, especially viral infection, is a very harmful clinical common disease. At first, people did not fully understand the molecular mechanism of the body's anti-viral infection. With the discovery of a class of cytokines - interferon, the natural immune cells and secondary acquired immune cells produced by the body against viral infection and their effects The molecular biological basis of the disease has gradually been understood. Interferon (IFN) is a general term for a family of cytokines with p...

Claims

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Application Information

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IPC IPC(8): C12N9/10C12N15/54A61K38/45A61P31/20A61P31/22A61P31/14
CPCA61K38/00C12N9/1007
Inventor 陈坤曹雪涛刘娟
Owner SECOND MILITARY MEDICAL UNIV OF THE PEOPLES LIBERATION ARMY
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