Marker of preeclampsia on gene level

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

Active Publication Date: 2018-10-26
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

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  • Marker of preeclampsia on gene level
  • Marker of preeclampsia on gene level
  • Marker of preeclampsia on gene level

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0062] Example 1 Screening of abnormally expressed genes in placental tissues of patients with preeclampsia

[0063] 1. Organize collection

[0064] A total of 5 cases of placental tissues from preeclampsia patients delivered in the hospital’s obstetrics and gynecology department were collected, and 5 cases of placental tissues from late-pregnancy women whose pregnancy was terminated due to social factors or abnormal pelvis during the same period were selected as normal control group. Multiple pregnancies, infectious diseases, chemical drug dependence, pregnant women’s smoking, fetal congenital malformations and other pregnancy complications and complications were excluded. The gestational week was strictly verified based on the B-ultrasound data of the first trimester, and the number of days of pregnancy was calculated. All the included subjects were Sign informed consent before collecting specimens. The diagnosis and classification criteria of preeclampsia refer to the seventh e...

Embodiment 2

[0101] Example 2 Large samples verify the expression of differentially expressed genes at the transcriptional level

[0102] 1. Organize collection

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

[0104] 2. Tissue RNA extraction and identification

[0105] The steps are the same as in Example 1.

[0106] 3. Design and preparation of primers

[0107] The primer sequences for qRT-PCR to detect PLEK gene expression and the primer sequences for qRT-PCR to amplify the internal reference GAPDH were designed and synthesized by Shenggong Bioengineering (Shanghai) Co., Ltd. After searching the UCSC database, their sequence information was entered into NCBI The software designs primers and confirms the correctness in the blast in the gene bank.

[0108] PLEK gene primer:

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

[0110] Downstream primer 5'-AACAGACTGGTTGGATAC-3...

Embodiment 3

[0133] Example 3 Promoting PLEK gene expression

[0134] 1. Construction of PLEK gene expression vector

[0135] According to the coding sequence of PLEK gene (as shown in SEQ ID NO. 5), the amplification primers are designed. 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 above cDNA sequence was double digested with restriction enzymes BamHI and XhoI and inserted into the restriction endonuclease. In the eukaryotic expression vector pcDNA3.1 digested with enzymes BamHI and XhoI, the recombinant vector pcDNA3.1-PLEK obtained by ligation was used in subsequent experiments.

[0136] 2. Cultivation of placental trophoblast cells

[0137] JEG-3 cells are placed in DMEM Gaotang medium containing double antibodies and 10% fetal bovine serum at 37℃, 5% CO 2 Cultured in the incubator, change the culture medium every 24h, and subculture once every 48h. Take the cells in the logarithmic ...

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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 present invention relates to the field of biomedicine. More specifically, the present invention relates to the application of PLEK gene in the preparation of products for the diagnosis and treatment of preeclampsia. Background technique [0002] Preeclampsia is one of the main diseases that cause maternal and child morbidity and mortality in the field of obstetrics. It has an obvious genetic tendency. Pathophysiological changes involve multiple systems and multiple organs. The etiology and pathogenesis are still not fully elucidated. , Has always been the focus of obstetric research. The change in gene expression is the core mechanism that regulates 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, Genes such as body fluid volume, placental growth, thrombosi...

Claims

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Application Information

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Patent Type & Authority Applications(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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