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Use of human homology cassette gene LHX4 in preparing medicine for treating phaeochromocytoma

A homeobox gene, pheochromocytoma technology, applied in the field of gene use, can solve the problems such as the inability to further study the function and the reduction of the expression level.

Inactive Publication Date: 2007-09-05
INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Since the expression level of the LHX4 gene was extremely reduced or disappeared after birth, and the mice after knockout of the LHX4 gene died, it was impossible to further study the functional role of the gene outside the regulation of neurodevelopment (1. Li, H., Witte, D.P., Banford, W. W., et al. Gsh-4 encodes a LIM-type homeodomain, is expressed in the developing central nervous system and is required for early postnatal survival. The EMBO J, 1994, 13: 2876-2885; 2. SinghG, Kaur S, Stock JL et al. Identification of 10 murine homeobox genes. Proc. Natl Acad. Sci. USA, 1991, 88: 10706-10710)

Method used

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  • Use of human homology cassette gene LHX4 in preparing medicine for treating phaeochromocytoma
  • Use of human homology cassette gene LHX4 in preparing medicine for treating phaeochromocytoma
  • Use of human homology cassette gene LHX4 in preparing medicine for treating phaeochromocytoma

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0042] Example 1 Expression of Human Homeobox Gene LHX4 in Different Adult Tissues

[0043] 1. Materials

[0044] Main instruments: HEAR hybrid hybridization oven (Heraeus Company), scanner (Kodak Company).

[0045] Main reagents: Huamn Ubiquitin Control cDNA Probes (Clontech, 5ng / μl), ExpressHyb hybridization solution (Clontech), salmon sperm DNA (10mg / ml, ssDNA, GIBCO-BRL), C 0 t-1 DNA (1mg / ml, Gibrco Company), 20×SSC (pH 7.0), 20% SDS (Sigma Company), washing solution 1: 2×SSC, 1% SDS, washing solution 2: 0.1×SSC , 0.5% SDS, Glass Milk Purification Kit (Yuanping Company), PCR Product Purification Kit (Promega Company), EX Taq PCR System (TaKaRa Company).

[0046] Hybridization membrane: MTE Array membrane (Multiple Tissue Expression Array, Clontech Company)

[0047] Autoradiography: α- 32 P dCTP (Beijing Yahui Company), X-ray film (Kodak Company), X-ray film press box (Shantou Medical Experimental Instrument Factory), D-72 developer, fixer (Zhejiang Nanhu Photographic I...

Embodiment 3

[0087] Example 3 Effect of Human Homeobox LHX4 Gene Transfection on PC12 Pheochromocytoma Cells

[0088] Effects on proliferation and differentiation in vitro

[0089] 1. Materials and methods

[0090] 1. Plasmid and cell line: pLXIN retrovirus expression vector, 6.1Kb, resistant to ampicillin and neomycin, purchased from Biological Reagent Company. SC712 plasmid, the plasmid containing the full-length CDS of the human homeobox gene LHX4 cloned for the first time in our laboratory, NIH Genbank accession number AF179849. PC12 cell line, a cell line cloned from male rat adrenal pheochromocytoma. There are NGF receptors on the membrane, and they can differentiate into mature dopaminergic sympathetic neurons after being induced by 50ng / ml NGF for 9 days.

[0091] 2. Construction of positive and antisense LHX4 retroviral plasmids (pLXIN-sense LHX4 and pLXIN-antisense LHX4): use restriction endonuclease site analysis software to analyze the restriction endonucleas...

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Abstract

The present invention discloses the application of human homology cassette gene LHX4 in preparing medicine for treating phaeochromocytoma. Through retroviral vector, human LHX4 gene is transduced into phaeochromocytoma cell PC12. Human LHX4 gene has functions of inhibiting the extracorporeal multiplication of PC12 and promoting the differentiation toward neure and axon growth of PC12, so that it may be used in preparing medicine for treating phaeochromocytoma.

Description

technical field [0001] The present invention relates to the use of a gene, in particular to the use of human homeobox gene LHX4 in preparing pheochromocytoma therapeutic drugs. Background technique [0002] Homeobox gene (homeobox gene) is a class of genes discovered in Gehring Laboratory in Switzerland in 1983. They share a 180-base-pair, highly conserved homeobox sequence at the 3′ end of the gene exon. Hence the name, the expression product of homeobox gene is an important type of transcription factor, the main control gene of development, and plays an important role in the regulation of embryonic development. The protein translated from this sequence is more conservative and rich in basic amino acids, so it is called homeodomain. LIM homeobox gene (LIM homeobox gene) belongs to the homeobox gene family, it not only has a homeobox structure, but also contains two LIM domains rich in cysteine ​​and histidine. The name of the LIM domain is derived from the acronym of the ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K48/00C12N15/12A61P35/00
Inventor 葛学铭范明吕艺刘淑红吴燕刘涛
Owner INST OF BASIC MEDICAL SCI ACAD OF MILITARY MEDICAL SCI OF PLA
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