ACADL gene and application of expression product of ACADL gene to preparation of abdominal aortic aneurysm diagnosis and treatment product

A technology for abdominal aortic aneurysm and gene expression, which is applied in the field of tumor diagnosis by detecting ACADL abnormality, and can solve the problem that the pathogenesis of abdominal aortic aneurysm is not completely clear

Active Publication Date: 2017-08-08
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the specific pathogenesis of abdominal aortic aneurysm is not completely clear. Existing reports have shown that its pathogenesis is closely related to factors such as genetic factors, inflammation, protease degradation, and smooth muscle cell apoptosis. different mutations

Method used

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  • ACADL gene and application of expression product of ACADL gene to preparation of abdominal aortic aneurysm diagnosis and treatment product
  • ACADL gene and application of expression product of ACADL gene to preparation of abdominal aortic aneurysm diagnosis and treatment product
  • ACADL gene and application of expression product of ACADL gene to preparation of abdominal aortic aneurysm diagnosis and treatment product

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Example 1 Gene Chip Screening for Differentially Expressed Genes

[0061] 1. Tissue acquisition and processing: Collect 2 live specimens of the middle layer of the AAA artery wall and 2 copies of the normal infrarenal aortic control tissue. Under sterile and RNase-free conditions, cut the aortic wall at about 4 cm below the flat renal artery. Rinse with sterilized normal saline to remove blood stains, remove arterial intima and intima from the collected tissues, and quickly (<5 min) freeze them in liquid nitrogen for later use.

[0062] 2. RNA extraction and cDNA synthesis

[0063] The aneurysm tissues preserved in liquid nitrogen were placed in a ceramic mortar under the low temperature environment of liquid nitrogen and crushed into powder, added Trizol reagent to homogenate, centrifuged, and the supernatant was extracted twice with 1:1 acidic phenol-chloroform After extraction with sodium acetate and 5:1 acidic phenol-chloroform, an equal volume of isopropanol preci...

Embodiment 2

[0072] Example 2 Verification of Differentially Expressed Genes in Large Samples

[0073] The ACADL that was expressed in abdominal aortic aneurysm tissue was selected as the research target for reverse validation.

[0074] 1. Tissue acquisition and processing: According to the method in Example 1, 30 live specimens of the middle layer of the AAA artery wall and 40 normal infrarenal aortic control tissues were collected.

[0075] 2. RNA extraction

[0076] RNA extraction was carried out according to the method of Example 1.

[0077] 3. Reverse transcription

[0078] 1 μg of total RNA was reverse-transcribed to synthesize cDNA using reverse transcription buffer. Use 25μl reaction system, take 1μg total RNA for each sample as template RNA, add the following components to the PCR tube respectively: DEPC water, 5× reverse transcription buffer, 10mmol / L dNTP, 0.1mmol / l DTT, 30μmmol / l Oligo dT, 200 U / μl M-MLV, template RNA. Incubate at 42°C for 1h, then centrifuge briefly at 72...

Embodiment 3

[0089] Example 3 overexpression of ACADL gene expression

[0090] 1. ACADL gene recombinant plasmid construction

[0091] (1) amplifying the coding sequence of the ACADL gene;

[0092] (2) Design amplification primers;

[0093] (3) The amplified ACADL gene was connected to the expression vector pcDNA3.0 to construct the pcDNA3.0-ACADL recombinant expression vector.

[0094] 2. Culture of human aortic smooth muscle cells

[0095] T / G HA-VSMC cells (referred to as HVSMC cells) were treated with DMEM high-glucose medium containing 10% fetal bovine serum (FBS) plus penicillin 100 units / ml and streptomycin 100 μg / ml, placed at 37°C, 5% CO 2 Cultured in an incubator, the culture medium was changed every 24 hours, and the cells were passaged once every 48 hours. Cells in the logarithmic growth phase were used for subsequent experiments.

[0096] 3. Cell transfection

[0097] Lipofectamine2000 was used as transfection reagent. The experiment was divided into two groups: negativ...

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Abstract

The invention discloses an ACADL gene and application of an expression product of the ACADL gene to preparation of an abdominal aortic aneurysm diagnosis and treatment product. As proved by gene chip and QPCR (Quantitative Polymerase Chain Reaction) experiments, compared with normal tissues, ACADL gene expression in abdominal aortic aneurysm tissues is different. A product for diagnosing abdominal aortic aneurysm is provided according to the relevance, and can be used for assisting in diagnosing the abdominal aortic aneurysm by detecting the level of the ACADL gene or the expression product of the ACADL gene in tissues. Moreover, the invention further provides an ACADL gene and an expression product of the ACADL gene, which can be taken as a target for developing an abdominal aortic aneurysm treating medicament in order to assist a clinician in making a personal treatment scheme.

Description

technical field [0001] The invention relates to the field of tumor diagnosis and treatment, more specifically, the invention relates to a tumor diagnosis method by means of detecting ACADL abnormality; and a tumor therapeutic agent for activating ACADL gene or protein. Background technique [0002] Abdominal aortic aneurysm (AAA) refers to a disease characterized by localized expansion and bulging of the abdominal aorta due to lesion or loss of the abdominal aortic wall, with a pulsating mass as the main symptom. AAA is usually defined as the continuous expansion of the three-layer structure of the abdominal aortic arterial wall to more than 1.5 times the diameter of the abdominal aorta at the renal artery. It is a common arterial degenerative disease with a high mortality rate. At present, the specific pathogenesis of abdominal aortic aneurysm is not completely clear. Existing reports have shown that its pathogenesis is closely related to factors such as genetic factors, in...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68G01N33/574A61K45/00A61P35/00
CPCA61K45/00C12Q1/6886C12Q2600/118C12Q2600/158G01N33/57407G01N33/57484G01N2800/52G01N2800/54
Inventor 李曙光孙锦云
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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