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Genetic markers of preeclampsia

A preeclampsia, gene technology, applied in the field of biomedicine, can solve problems such as abnormal expression of multiple genes

Active Publication Date: 2020-06-09
QINGDAO MEDINTELL BIOMEDICAL CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although many preeclampsia-related genes have been found, such as angiotensinogen (AGT) gene (Ward K, Hata A, Jeunemaitre X, et al. A molecular variant of angiotensiongen associated with preeclampsia. Nature Genet, 1993, 4:59 -61.), prothrombin gene [Kupferminc MJ, Eldor A, Steinman N, et al.Increased frequency of genetic thrombophilia in women with complications of pregnancy.N Engl J Ned,1999,340:9-13], tumor necrosis factor a(TNF a) gene [Rinehart BK, Terrone DA, Lagoo-Deenadayalan S, et al. Expression of the placental cytokines tumor necrosis factor alpha, interleukin 1beta, and interlukin 10 is increased in preeclampsia. Am J Obstet Gynecol, 1999, 181(4) :915-920.], etc., but because its occurrence and development involve abnormal expression of multiple genes, more in-depth research is still needed

Method used

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  • Genetic markers of preeclampsia
  • Genetic markers of preeclampsia
  • Genetic markers of preeclampsia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Screening for Abnormally Expressed Genes in the Placental Tissue of Patients with Preeclampsia

[0063] 1. Organization collection

[0064] The placental tissues of 5 patients with preeclampsia who were delivered in the obstetrics and gynecology department of the hospital were collected, and 5 cases of placental tissues of late-pregnancy pregnant women with normal blood pressure who underwent surgical termination of pregnancy due to social factors or pelvic abnormalities were selected as the normal control group. Multiple pregnancy, infectious diseases, chemical drug dependence, maternal smoking, fetal congenital malformations and other pregnancy complications and complications were all excluded, and the gestational age was strictly verified according to the B-ultrasound data in the first trimester, and the number of days of pregnancy was calculated. Sign the informed consent form before collecting the specimens. The diagnosis and grading standards of preecla...

Embodiment 2

[0101] Example 2 Large sample verification of expression of differentially expressed genes at the transcriptional level

[0102] 1. Organization collection

[0103] According to the standard of Example 1, 35 cases of placental tissues of preeclampsia patients and 40 cases of placental tissues of normal pregnant women were collected.

[0104] 2. Tissue RNA extraction and identification

[0105] Step is with embodiment 1.

[0106] 3. Design and preparation of primers

[0107] The primer sequences for qRT-PCR detection of PLEK gene expression and the primer sequences for qRT-PCR amplification of the internal reference GAPDH were designed and synthesized by Sangon Bioengineering (Shanghai) Co., Ltd., and their sequence information was entered into NCBI after searching the UCSC database The primers are designed by the software and confirmed by blast in the gene bank.

[0108] PLEK gene primers:

[0109] Upstream primer: 5'-ATTGACTTAGGTGCCTTAT-3' (SEQ ID NO.1);

[0110] Downst...

Embodiment 3

[0133] Embodiment 3 promotes PLEK gene expression

[0134] 1. Construction of PLEK gene expression vector

[0135] Amplification primers were designed according to the coding sequence of the PLEK gene (as shown in SEQ ID NO.5). The coding sequence of the full-length PLEK gene was amplified from the cDNA library of adult fetal brain (clontech company, catalog number: 638831). The obtained recombinant vector pcDNA3.1-PLEK was connected to the eukaryotic cell expression vector pcDNA3.1 double-digested with enzymes BamHI and XhoI for subsequent experiments.

[0136] 2. Culture of placental trophoblast cells

[0137] JEG-3 cells were placed in DMEM high pond medium containing double antibody and 10% fetal bovine serum 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.

[0138] 3. Cell transfection

[0139] Lipofect...

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Abstract

The invention discloses a diagnosis and treatment target-PLEK gene of preeclampsia. Whether a subject has the preeclampsia is judged or whether the subject has the risk of being suffered from preeclampsia is diagnosed through detecting the content of a PLEK gene and an expression product thereof in a placental tissue of the subject. Furthermore, a proliferation index of a preeclampsia cell, whichis cultured in vitro, is researched and proves that the PLEK gene is used as a medicine target for treating the preeclampsia.

Description

technical field [0001] The invention relates to the field of biomedicine, more specifically, the invention relates to the application of PLEK gene in the preparation of products for diagnosis and treatment of preeclampsia. Background technique [0002] Preeclampsia is one of the main diseases that cause increased maternal and child morbidity and mortality in the field of obstetrics. It has obvious genetic predisposition, and its pathophysiological changes involve multi-system and multi-organ lesions. The etiology and pathogenesis have not yet been fully elucidated. , has always been the focus of obstetrics research. The change of gene expression is the core mechanism to regulate the process of cell life activities. Abnormal metabolic processes or pathological changes, whether controlled by a single gene or a multi-gene system, are essentially caused by differential expression of genes, so they can regulate blood pressure, The genes of body fluid volume, placental growth, th...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/6883G01N33/68A61K45/00A61P25/08
CPCA61K45/00A61P25/08C12Q1/6883G01N33/68G01N2800/2857
Inventor 孙耀兰张冬梅
Owner QINGDAO MEDINTELL BIOMEDICAL CO LTD
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