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Long non-coding RNA (ribonucleic acid) and application thereof in diagnosing/treating preeclampsia

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

Active Publication Date: 2020-05-19
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 (ribonucleic acid) and application thereof in diagnosing/treating preeclampsia
  • Long non-coding RNA (ribonucleic acid) and application thereof in diagnosing/treating preeclampsia
  • Long non-coding RNA (ribonucleic acid) and application thereof in diagnosing/treating preeclampsia

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0077] Detect the expression of PDIA3P1 in tissues and cells

[0078] Take 0.1g tissue, grind it sufficiently with liquid nitrogen (into a powder form) or discard the culture medium for 1-5×107 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 of 75% ethanol (p...

Embodiment 2

[0100] To study the effect of PDIA3P1 on the phenotype of normal trophoblast HTR-8 / SVneo and JEG3 cells.

[0101] First, the normal trophoblast HTR-8 / SVneo cell line was selected as the research object of this experiment, using lip2000 as the carrier, transfected with PDIA3P1 interference sequence to knock down the expression of PDIA3P1 to simulate the pathogenesis of preeclampsia, MTT and clonal proliferation assay It was found that the interfering sequence of PDIA3P1 was transfected in HTR-8 / SVneo cells, and the cell growth was inhibited after knocking down the expression of PDIA3P1. Thus, these data indicate that PDIA3P1 can promote the proliferation ability of HTR-8 / SVneo cells.

Embodiment 3

[0103] Effect of PDIA3P1 on apoptosis of placental trophoblast cells HTR-8 / SVneo.

[0104] In order to investigate whether PDIA3P1 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 PDIA3P1 interference sequence to knock down the expression of PDIA3P1 Simulate the pathogenesis of preeclampsia. We performed flow cytometry to analyze whether apoptosis was involved in the inhibition of cell growth induced by PDIA3P1 knockdown. Such as figure 2 As shown in C, early apoptosis (UR) and late apoptosis rate (LR) were higher in PDIA3P1 knockdown HTR-8 / SVneo cells than in control cells. Thus, PDIA3P1 inhibits the apoptosis of trophoblasts.

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Abstract

The invention belongs to the field of gene engineering and in particular relates to application of PDIA3P1 in diagnosing preeclampsia and preparing medicines for target treatment. Reduction of PDIA3P1in placenta tissue of a pregnant woman suffering from preeclampsia is related to occurrence and development of preeclampsia, PDIA3P1 of a low expression level is closely related to pathogenesis of preeclampsia, by changing expression of PDIA3P1, proliferation, apoptosis, invasion, migration and the like of trophocyte of the pregnant woman suffering from preeclampsia can be affected, and by improving expression of PDIA3P1, proliferation, invasion and migration of the trophocyte can be promoted.

Description

technical field [0001] The invention belongs to the field of genetic engineering, and particularly relates to the application of long non-coding RNA-PDIA3P1 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 st...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/113C12Q1/6883A61K31/7088A61P15/00
CPCC12N15/113C12Q1/6883A61K31/7088A61P15/00C12N2310/14C12N2310/10C12Q2600/158C12Q2600/178Y02A50/30
Inventor 许叶涛夏曦张媛媛汤卫春
Owner JIANGSU PROVINCE HOSPITAL THE FIRST AFFILIATED HOSPITAL WITH NANJING MEDICAL UNIV
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