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Novel application of HIP-55 (hematopoietic progenitor kinase 1 [HPK1]-interacting protein of 55kDa)
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A kind of HIP-55, 1. HIP-55 technology, applied in the field of new application of HIP-55, can solve the problems of less understanding of HIP-55 functions, HIP-55 research reports, etc.
Inactive Publication Date: 2013-05-22
PEKING UNIV THIRD HOSPITAL
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Problems solved by technology
At present, the function of HIP-55 is poorly understood, and there is no research report on the role of HIP-55 in the cardiovascular system.
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Embodiment 1
[0040] Example 1. Obtaining of HIP-55shRNA, recombinant adenovirus overexpressing HIP-55 and recombinant adenovirus expressing RNA interference HIP-55
[0041] 1. Obtaining HIP-55shRNA
[0042] The following HIP-55 shRNA and control shRNA were designed and synthesized for the mouse HIP-55 protein coding gene as shown in Table 1:
[0043] HIP-55shRNA, its nucleotide sequence is sequence 3 in the sequence listing, and the nucleotide sequence of its coding gene is sequence 4 in the sequence listing;
[0044] Design Control shRNA: its nucleotide sequence is sequence 5 in the sequence listing, and the nucleotide sequence of its coding gene is sequence 6 in the sequence listing;
[0045] Table 1 is shRNA and its control sequence
[0046]
[0047] 2. Construction and preparation of recombinant adenovirus overexpressing HIP-55
[0048] 1. Construction of recombinant adenoviral vector for overexpressing HIP-55
[0049] Amplification of attB1-HIP-55-Flag-IRES / DesRed-attB2 by Overla...
Embodiment 2
[0185] Example 2, Functional Research of HIP-55
[0186] 1. The effect of HIP-55 on the proliferation of cardiac fibroblasts
[0187] 1) Brdu ELISA detection
[0188] The isolated cardiac fibroblasts were subcultured in a 96-well plate with a cell density of 3.5*10 4 cells / mL, 100 μl / well, when the cell confluency reaches 60-70%, replace with serum-free culture, and infect with adenovirus; the above-mentioned adenoviruses are the overexpressed HIP-55 recombinant adenovirus prepared in Example 1 respectively ( Dsred-HIP55), Des1d adenovirus (Dsred), shRNA interference HIP-55 expression recombinant adenovirus (KD1), control shRNA interference recombinant adenovirus (scramble), pAd / PL-Dest adenovirus. After culturing for 24 hours, give isoproterenol (ISO, the final concentration is 10 -5 mol / L) were collected after 24 hours of treatment. Six hours before sample collection, 10 μl / well of BrdU was added (final concentration 10 μmol / L). Discard the supernatant when collecting s...
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Abstract
The invention discloses a novel application of HIP-55 (hematopoietic progenitorkinase 1 [HPK1]-interacting protein of 55kDa). The invention provides an application of an HIP-55 protein, an HIP-55 protein coding gene or a recombinant vector containing the HIP-55 protein codinggene or adenovirus containing the HIP-55 protein codinggene to preparation of products with the function (1) or (2): (1) suppression of cardiac fibroblasts proliferation; and (2) suppression of P38MAPK (P38 mitogen activated proteinkinase) activity; and an amino acid sequence of the HIP-55 protein is a sequence 2 in a sequence table. Experiments prove that the HIP-55 has expressions in different organs of a rat, especially has a high-level expression in the heart and further has an overexpression in cardiac fibroblasts of a small rat under the induction of ISO (isoprenaline), and therefore the HIP-55 can be used for remarkably suppressing the cardiac fibroblast proliferation; and the HIP-55 can be used for suppressing P38MAPK activation induced by ISO and the kinase activity in the overexpression of HIP-55 cardiac fibroblasts.
Description
technical field [0001] The invention relates to the field of biotechnology, in particular to a new application of HIP-55. Background technique [0002] HIP-55 (hematopoietic progenitorkinase 1 [HPK1]-interacting protein of 55kDa; also known as drebrin-like protein, mAbp1 or SH3P7) is a protein containing a multifunctional domain. Its structure mainly includes the N-terminal F-actin protein binding domain and the C-terminal SH3 domain. HIP-55 is involved in synaptogenesis, receptorendocytosis, cytoskeleton rearrangement and signal transduction. At present, little is known about the function of HIP-55, and there is no research report on the role of HIP-55 in the cardiovascular system. [0003] Cardiovascular disease is the "number one killer" of Chinese residents' health, and the annual direct medical expenses mainly for cardiovascular disease have reached 130 billion yuan. Myocardial fibrosis is an important pathological change of cardiac remodeling. It refers to the exc...
Claims
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