Target for resisting hepatitis b virus and application of the target

A hepatitis B virus and target technology, applied in the field of molecular biology and biomedicine, to achieve the effect of a wide range of medical application prospects

Active Publication Date: 2017-11-03
AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Chinese patent CN105525018A discloses the application of FAM176A gene in the diagnosis and treatment of rectal adenocarcinoma, which shows that increasing the expression of FAM176A can inhibit the proliferation of rectal adenocarcinoma cells and promote the apoptosis of rectal adenocarcinoma cells. related reports

Method used

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  • Target for resisting hepatitis b virus and application of the target
  • Target for resisting hepatitis b virus and application of the target
  • Target for resisting hepatitis b virus and application of the target

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] This example is to verify the effect of FAM176A on the HBV replication process in a cell experiment.

[0027] In Huh7 cells at 4×10 5 After overnight culture, co-transfect pHBV 1.3 plasmid and different doses of FAM176A-Flag plasmid, collect cells and cell supernatant 5 days after transfection. Use Northern blot method to detect HBV RNA, use Southern blot method to detect HBV DNA in cell lysate, use non-denaturing gel to detect HBV nucleocapsid (corecapsid) formation, use ELISA kit to detect HBsAg and HBeAg in cell supernatant, and use Western The blot method was used to detect the expression of HBV core protein (HBV core protein) and surface antigen (S protein).

[0028] Such as figure 1 As shown, the experimental results showed that after overexpressing FAM176A in Huh7 cells, the products during HBV replication were significantly reduced, including HBV RNA, HBV DNA, HBsAg and HBeAg in the cell supernatant, HBV core protein and S protein, and showed Dose-dependent, ...

Embodiment 2

[0030] This example is to verify the effect of FAM176A on the HBV replication process in animal experiments.

[0031] In previous studies, the pAAV / HBV1.2 plasmid was injected into the tail vein of hydrodynamic high pressure to construct a mouse model of chronic HBV infection. C3H / HeN mice had a higher success rate of modeling, and the maintenance time of serum HBsAg and HBV DNA in mice was longer. Therefore, we established a chronic HBV infection model in C3H / HeN mice by hydrodynamic high pressure tail vein injection in Shanghai Public Health Center.

[0032] And based on this model, the Fam176a-Flag plasmid and the pAAV / HBV1.2 plasmid were simultaneously injected into mice by hydrodynamic high-pressure tail vein injection. Serum was collected at 3d, 1w, 2w, 3w, 4w, 5w, 6w, 7w, and 8w after injection, and the HBsAg and HBeAg in the serum were detected by the Abbott kit, and the HBV DNA in the serum was detected by the Realtime PCR method (the detection limit was 500IU / ml)....

Embodiment 3

[0035] This example is the prediction of the secondary structure of the FAM176A protein, and the prediction steps are as follows:

[0036] Get the website of the predicted protein domain from the previous literature:

[0037] http: / / www.cbs.dtu.dk / services / TMHMM-2.0 /

[0038] http: / / bioinf.cs.ucl.ac.uk / psipred /

[0039] http: / / www.sbg.bio.ic.ac.uk / phyre2 / html / page.cgi? id=index

[0040]The protein sequence encoded by the FAM176A gene was obtained from Pubmed (http: / / www.ncbi.nlm.nih.gov / pubmed / ) website. According to the website of the predicted protein secondary structure verified in the previous literature, after inputting the amino acid sequence of the FAM176A protein, the prediction result of the secondary structure of the FAM176A protein was obtained.

[0041] Such as image 3 As shown, the FAM176A protein is a transmembrane protein, its 1-34 amino acid is located outside the membrane, the 35-57 amino acid is the transmembrane region, and the 58-152 amino acid is the...

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Abstract

The invention discloses application of a gene FAM176A of a liver tissue and / or protein FAM176A coded by the gene FAM176A in serving as a target for resisting a hepatitis b virus or application of the gene or the protein in preparation of an anti-hepatitis b virus drug. Membrane-spanning protein FAM176A coded by the gene FAM176A of the liver tissue can inhibit duplication of HBV. An in vitro cell experiment and an in vivo animal experiment show that the FAM176A can inhibit the duplication of the HBV, and the structure of the protein FAM176A is forecast; moreover, a membrane-spanning region of the protein FAM176A is a key functional region for inhibiting the duplication of the HBV, which provides a new target for further development of a new anti-HBV drug; the target has a very wide medical application prospect.

Description

technical field [0001] The present invention relates to the technical fields of molecular biology and biomedicine, in particular to an anti-hepatitis B virus target and its application. As a new anti-HBV target. Background technique [0002] Chronic hepatitis B virus (Hepatitis B Virus, HBV) infection is a major global health problem. The HBV infection rate is high in my country. The latest cross-sectional study shows that the positive rate of HBsAg among men aged 21-49 in rural my country is 6%. In the natural course of chronic HBV infection, according to the indicators of serum hepatitis B markers, HBV viral load (HBV DNA) and liver tissue inflammation, the natural history can be divided into four periods: immune tolerance period, immune clearance period, non Active or low (non-)replication periods and reactivity periods ( figure 1 ). The goal of treatment in patients with chronic HBV infection is to inhibit HBV replication before significant liver damage occurs. At p...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/68A61K48/00A61K38/17A61P31/20A61P1/16
CPCA61K38/1709A61K48/005C12Q1/6883C12Q2600/106G01N33/68
Inventor 张继明陆蒙吉刘红艳于洁沈忠良
Owner AFFILIATED HUSN HOSPITAL OF FUDAN UNIV
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