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Long non-coding RNA and application thereof in diagnosis/treatment of preeclampsia

A preeclampsia, non-coding technology, applied in the field of genetic engineering, can solve the problem of incomplete function research of lncRNAs

Active Publication Date: 2020-05-15
JIANGSU PROVINCE HOSPITAL THE FIRST AFFILIATED HOSPITAL WITH NANJING MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Recently, the role of miRNAs in various aspects of cellular processes has been confirmed, however, the function of lncRNAs is not very thorough

Method used

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  • Long non-coding RNA and application thereof in diagnosis/treatment of preeclampsia
  • Long non-coding RNA and application thereof in diagnosis/treatment of preeclampsia
  • Long non-coding RNA and application thereof in diagnosis/treatment of preeclampsia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0092] Example 1 Detection of the expression of AGAP2-AS1 in tissues and cells

[0093] Take 0.1g tissue, grind it sufficiently with liquid nitrogen (into powder) or 1-5×10 7 Discard the culture medium from the cells, and rinse twice with pre-cooled PBS. Add 1ml of Trizol lysate, pipette and mix with an enzyme-free pipette tip, let stand for 5min, and transfer the lysate into a pre-labeled 1.5ml enzyme-free centrifuge tube. Centrifuge at 7500 g at 4°C for 5 minutes, take the supernatant and add 1 / 5 volume of chloroform, invert and mix for 30 seconds, and let stand for 2 minutes. Centrifuge at 12000g for 15min at 4°C. The solution is divided into three layers (aqueous phase-white precipitate-red organic matter), transfer the aqueous phase layer to a new 1.5ml centrifuge tube, try not to absorb the white precipitate. Add an equal volume of isopropanol, mix gently by inversion, and let stand for 5-10min. Centrifuge at 12000g for 10min at 4°C. Discard the supernatant, add 1ml...

Embodiment 2

[0116] To study the effect of AGAP2-AS1 on the phenotype of normal trophoblast HTR-8 / SVneo and JEG3 cells.

[0117] First, the normal trophoblast HTR-8 / SVneo cell line was selected as the research object of this experiment, and lip2000 was used as the carrier to transfect the AGAP2-AS1 interference sequence to knock down the expression of AGAP2-AS1 to simulate the pathogenesis of preeclampsia, MTT and The BrdU proliferation assay found that the interfering sequence of AGAP2-AS1 was transfected in HTR-8 / SVneo cells, and the cell growth was inhibited after knocking down the expression of AGAP2-AS1. In contrast, the AGAP2-AS1 plasmid was transfected in normal trophoblast cells to further verify the functional role of AGAP2-AS1, and found that the overexpression of AGAP2-AS1 promoted the proliferation of HTR-8 / SVneo cells ( figure 2 A-D). Thus, these data suggest that AGAP2-AS1 can promote the proliferation ability of HTR-8 / SVneo cells.

Embodiment 3

[0119] Effects of AGAP2-AS1 on cycle and apoptosis of placental trophoblast cells HTR-8 / SVneo and JEG3

[0120] In order to investigate whether AGAP2-AS1 affects cell cycle transition on the proliferation of HTR-8 / SVneo cells, the normal trophoblast HTR-8 / SVneo cell line was used as the research object, and lip2000 was used as the vector to transfect the AGAP2-AS1 interference sequence to knock out Low AGAP2-AS1 expression mimics the pathogenesis of preeclampsia. Cell cycle analysis by flow cytometry. The results showed that after HTR-8 / SVne o cells were transfected with the AGAP2-AS1 interference sequence, it was found that the cell cycle was arrested in the G1 / G0 phase after knocking down the expression of AGAP2-AS1. The proportion of stagnation in G1 / G0 phase was lower than that of the control group ( figure 2 E-F, P figure 2 As shown in G and 2H, early apoptosis (UR) and late apoptosis rate (LR) were higher in AGAP2-AS1 knockdown HTR-8 / SVneo cells than control cells. T...

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Abstract

The invention belongs to the genetic engineering field, and particularly relates to application of AGAP2-AS1 in the preparation of target drugs for diagnosing and treating preeclampsia. Down regulation of AGAP2-AS1 in placenta tissues of a pregnant woman in the preeclampsia is related with the occurrence and development of the preeclampsia, AGAP2-AS1 in a low expression level is in a close relationship with pathogenesis of the preeclampsia, the proliferation, apoptosis, invasion, migration and the like of trophoblastic cells of the pregnant woman in the preeclampsia are influenced through changing the expression of AGAP2-AS1, and the proliferation, invasion and migration of the trophoblastic cells can be promoted through the enhancement of the expression of AGAP2-AS1.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and particularly relates to the application of long non-coding RNA-AGAP2-AS1 in diagnosis and preparation of drugs for treating preeclampsia. Background technique [0002] Preeclampsia is one of the most common pregnancy complications in the world, and it is the main cause of death related to pregnancy complications. According to the statistics of the World Health Organization, preeclampsia or eclampsia directly leads to about 14% (about 8500) of the world's annual Maternal death, which is one of the most common causes of preterm birth and fetal growth restriction and maternal death. Although the current medical treatment has been greatly developed, but the overall incidence of patients is still high. With the rapid development of sequencing technology and molecular biology, gene diagnosis and molecular targeted therapy have become a hot issue in the treatment of preeclampsia. Therefore, the ...

Claims

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Application Information

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IPC IPC(8): C12Q1/6883C12N15/11A61K31/7088A61P15/00
CPCC12Q1/6883A61K31/7088A61P15/00C12Q2600/158C12Q2600/178
Inventor 杨娜娜许叶涛张媛媛吴丹
Owner JIANGSU PROVINCE HOSPITAL THE FIRST AFFILIATED HOSPITAL WITH NANJING MEDICAL UNIV
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