CN112210599B - Long non-coding RNA for preeclampsia clinical risk assessment - Google Patents
Long non-coding RNA for preeclampsia clinical risk assessment Download PDFInfo
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Abstract
A long non-coding RNA for use in preeclampsia clinical risk assessment comprising the steps of: looking up and analyzing medical record data of the patient, and collecting placenta tissues of normal pregnant women and preeclamptic pregnant women; preparing a placenta tissue paraffin section, and staining a placenta tissue masson pine to observe the change of a placenta tissue structure; bioinformatics analysis of the structure of the damage-induced non-coding RNA; extracting long-chain non-coding RNA of placenta tissue, and detecting the change of the expression of the non-coding RNA induced by the injury; culturing human placenta trophoblasts, and screening damage-induced non-coding RNA interference fragments; detecting a change in matrix metalloproteinase 2 (MMP-2) protein expression following interference with the injury-induced non-coding RNA; after interfering with the non-coding RNA induced by the injury, a scratch test is used for detecting the migration capability of the cells and an invasion test is used for detecting the invasion change of the cells. The invention obtains materials from placenta tissues, is simple and convenient to obtain samples, does not cause any wound of patients and has stronger feasibility.
Description
Technical Field
The invention belongs to the technical field of medicine, and relates to a long-chain non-coding RNA for preeclampsia clinical risk assessment.
Background
Preeclampsia (PE) is a common complication in pregnancy causing multiple organ and multiple system injuries of pregnant and lying-in women, belongs to one of special diseases in pregnancy, usually presents hypertension and proteinuria after 20 weeks of pregnancy as main clinical manifestations, can generate general system dysfunction in severe cases, and is an important reason for death of pregnant and lying-in women and perinatals. Blood, urine, eye ground examination and the like are common diagnosis methods at the present stage of preeclampsia, but the methods are still mainly used for diagnosing clinical manifestations of diseases by means of characteristics traditionally, but generally, the gestational week is large when the diagnosis is clear. In the aspect of treatment, symptoms can be only alleviated by symptomatic treatment at present, and due to the particularity of the medication of pregnant women, few medicines can ensure the safety and the curative effect; in the absence of an effective treatment regime, termination of pregnancy is the only means of a complete cure, and the delivery of preterm and low birth weight infants will place a heavy economic burden on the family. Based on the above, a molecular diagnostic marker for preeclampsia clinical risk assessment is found, a prevention and treatment strategy for preeclampsia is discussed, and a national 'good birth and good care' policy is responded comprehensively, so that the molecular diagnostic marker has long-term and profound significance for improving human reproductive health level and optimizing birth population quality.
The etiology of preeclampsia may involve a variety of factors, including abnormal trophoblast infiltration, immunoregulatory dysfunction, genetic factors, insulin resistance, and malnutrition, among others. At present, the uterine spiral artery remodeling disorder caused by too shallow infiltration of trophoblasts and the shallow implantation of placenta are widely considered to be the main reasons for the occurrence and development of preeclampsia. The development of preeclampsia is mainly divided into two stages, the early gestation stage: under the action of multiple factors such as insufficient blood perfusion, immune tolerance and the like in the formation period of the placenta, the invasion of trophoblasts is insufficient, and the recasting of spiral arteries is obstructed, so that the blood and oxygen supply of the placenta are insufficient; in the late stage of pregnancy: because of oxidative stress, inflammatory reaction and the like, the placenta synthesizes and releases a large amount of factors, thereby causing dysfunction of maternal vascular endothelium, dysfunction of endothelial cells of various organs and systemic small vasospasm, and further causing various clinical symptoms. The placenta is an important temporary organ specific to gestation, bears important functions of maternal nutrition transmission, maternal-fetal barrier and the like, is also an important link for adaptive regulation between pregnancies in gestation, and abnormal development of the placenta is a key for poor pregnancy fate such as preeclampsia and the like. The trophoblast is used as an important functional cell of placenta tissue, the original trophoblast is gradually differentiated into an extravillous trophoblast during normal pregnancy, the cell is proliferated and invaded to promote helical artery remodeling, and the cell penetrates through a maternal-fetal interface through the infiltration capacity similar to that of a tumor cell to promote the formation of a placenta, so that the normal placenta function is maintained. Therefore, the proliferation and invasion capacity of the trophoblasts outside the villi is critical to the development of the placenta.
Long non-coding RNAs (lncrnas) are a class of endogenous RNA molecules with a length greater than 200 nucleotides, located in cytoplasm or nucleus, and have no function of coding proteins due to lack of significant Open Reading Frames (ORFs), and in recent years, with the development of high-throughput sequencing and bioinformatics technologies, detection finds that more and more long non-coding RNAs can influence the occurrence of diseases by regulating and controlling processes such as chromatin remodeling, mRNA degradation and translation inhibition. The literature reports that long-chain non-coding RNA shows different expression modes in the occurrence and development of preeclampsia, plays different roles, and can influence the proliferation, migration, invasion, apoptosis and the like of trophoblasts so as to influence the occurrence and development of preeclampsia. Damage-Induced non-coding RNA (DINO) is a member of long-chain non-coding RNA molecules, can promote DNA Damage and influence biological processes such as apoptosis, and has the possibility of participating in the occurrence and development of preeclampsia through regulating and controlling cell invasion and migration. According to the application, the expression of lncRNA DINO (damage-induced non-coding RNA) in the placenta tissue of a preeclamptic patient is increased through RT-qPCR detection in the placenta tissue of the preeclamptic patient, lncRNA DINO interference fragments are transfected at a cell level, cell migration and invasion capacity are enhanced through scratch experiments and Transwell invasion experiments, and MMP-2 protein expression is increased, so that the increase of lncRNA DINO expression can reduce the migration and invasion capacity of trophoblasts.
Disclosure of Invention
The invention aims to provide a long-chain non-coding RNA for preeclampsia clinical risk assessment, the long-chain non-coding RNA and action verification material selection are respectively from placenta tissues and cells, the sample acquisition mode and the sample acquisition way are relatively simple and quick, no wound is caused, the long-chain non-coding RNA is generally accepted by patients, and the long-chain non-coding RNA has high feasibility.
The specific technical scheme is as follows:
a long non-coding RNA for use in preeclampsia clinical risk assessment comprising the steps of:
(1) Looking up and analyzing medical record data of the patient, and collecting placenta tissues of normal pregnant women and preeclamptic pregnant women;
(2) Preparing a placenta tissue paraffin section, and staining a placenta tissue masson pine to observe the change of a placenta tissue structure;
(3) Bioinformatics analysis of the structure of the lesion-induced non-coding RNA;
(4) Extracting long-chain non-coding RNA of the placenta tissue, and detecting the change of the expression of the non-coding RNA induced by the injury;
(5) Culturing human placenta trophoblasts, and screening damage-induced non-coding RNA interference fragments;
(6) Detecting a change in MMP-2 (matrix metalloproteinase 2) protein expression following interference with injury-induced non-coding RNA;
(7) After interfering with the non-coding RNA induced by the injury, a scratch test is used for detecting the migration capacity of the cells and an invasion test is used for detecting the invasion change of the cells.
Compared with the prior art, the invention has the beneficial effects that:
1. the invention defines that long-chain non-coding RNA (lncRNA DINO) is used as a key molecule for early warning and diagnosis of preeclampsia.
2. The sample material is obtained from placenta tissue, the sample is easy and convenient to obtain, no wound is caused to the patient, the understanding of the patient can be generally realized, and the feasibility is high.
Drawings
FIG. 1 is a chart of clinical data of pregnant patients.
FIG. 2 is a schematic illustration of the staining of placental tissue by Masson.
FIG. 3 is a diagram illustrating the structure and position of a long non-coding RNA in bioinformatics analysis.
FIG. 4 is a schematic representation of the expression of long non-coding RNA from placental tissue.
FIG. 5 is a schematic diagram of the selection of injury-induced interfering fragments of non-coding RNA.
FIG. 6 is a schematic diagram showing the change in expression of MMP-2 and mRNA after disruption of injury-induced non-coding RNA.
FIG. 7 is a schematic representation of the change in MMP-2 protein expression following interference with injury-induced non-coding RNA.
FIG. 8 is a schematic diagram showing the detection of cell migration ability by scratch test and the detection of cell invasion change by invasion test after interfering with injury-induced non-coding RNA.
In FIG. 1, the collected clinical history data of preeclamptic patients includes pregnancy week, height, weight, BMI, blood pressure, urine protein, etc.; note: * P<0.05, compared to the normal pregnancy group.
In FIG. 2, the NC group was the normal pregnancy group and the PE group was the preeclampsia group.
FIG. 3, wherein (A) the position of DINO on the chromosome; (B) Gene source and related information for DINO (damage-induced non-coding RNA); (C) transcript length of DINO.
In fig. 4, ordinate: lncRNA DINO expression levels are the expression of lncRNA DINO (DNA damage induced non-coding RNA); the abscissa: NC represents a normal pregnancy group, and PE represents a preeclampsia group; note: * P<0.01, compared to the normal pregnancy group.
In fig. 5, ordinate: lncRNA DINO expression levels are the expression of non-coding RNA induced by DNA damage; the abscissa: 0 represents a normal cell group, si-NC represents a transfection negative control interference fragment cell group, and si-DINO represents non-coding RNA induced by transfected DNA damageA set of interfering fragment cells; note: * P<0.05, compared to a blank control group.
In fig. 6, ordinate: MMP-2 (matrix metalloproteinase 2) mRNA (messenger single-stranded ribonucleic acid) expression level represents the expression level of the messenger single-stranded ribonucleic acid of the matrix metalloproteinase 2; the abscissa: si-NC represents the group of interference fragments of transfection negative control, si-DINO represents the group of interference fragments of transfection DNA damage-induced non-coding RNA; note: * P<0.05, compared to the blank control group.
In fig. 7, ordinate: MMP-2 mRNA expression level represents the expression level of the mRNAs of the matrix metalloproteinase 2; the abscissa: si-NC represents the group of interference fragments of transfection negative control, si-DINO represents the group of interference fragments of transfection DNA damage-induced non-coding RNA; note: * P<0.05, compared to the blank control group.
In fig. 8, a. Cell invasion experiment, b. Cell scratch experiment; ordinate: invaded cells represents the number of invading cells; the si-NC represents the interference fragment group of the transfection negative control, and the si-DINO represents the interference fragment group of the transfection DINO; note: * P<0.05, compared to the blank control group.
Detailed Description
The technical scheme of the invention is explained in detail by combining the attached drawings and the specific embodiment.
The long-chain non-coding RNA for preeclampsia clinical risk assessment specifically comprises the following steps:
1. material preparation
1.1 Materials and reagents: human placental trophoblast line HTR-8/SVneo (human choriotrophoblast cells) (China, shanghai Rick Biotech Co., ltd.); fetal bovine serum, RPMI1640 medium (australia, gibco); streptomycin, trypsin digestive juice, NP-40 (China, biyuntian biotechnological research institute); masson staining kit, PMSF (china, beijing solibao science and technology ltd); MMP-2 (matrix metalloproteinase 2) antibody (Abcam, uk), β -actin (β -actin) antibody, horseradish peroxidase-labeled secondary antibody (china, beijing bridge ltd); skimmed milk powder (us, BD company); a cDNA kit, RT-qPCR (real-time quantitative fluorescent PCR) kit (Japan, takara Co.); lipo2000 (Thermo Fisher, usa); the primers were synthesized by Shanghai Biotech, inc.; lncRNA DINO siRNA (injury-induced interfering RNA fragment of non-coding RNA) was constructed by hippo, lakezhou.
1.2 Instruments and equipment clean bench (suzhou, santai); CO 2 2 Incubators (Heraeus, germany); model 5415D micro bench centrifuge (Eppendorf, germany); ultramicrospectrophotometers (simplina, usa); vertical electrophoresis apparatus, membrane transfer apparatus, exposure apparatus (Bio-Rad, USA); BS110S precision balance (Sartorius, germany); horizontal shaker (Thermo Fisher, usa) gradient RCR instrument (Biometra Tone, germany); fluorescent quantitative PCR instrument (Shanghai, feng Ling); laser confocal (japan, nikon).
2. Method of implementation
2.1 selection of clinical specimens
In the study, 20 preeclamptic pregnant women born by caesarean section in the Ningxia Longfu hospital from 2019 and 6 to 2019 are selected as a PE (preeclampsia) group; in addition, 20 normal pregnant women who are born by caesarean section at the same time are randomly selected as an NC group. All pregnant women are in single birth and initial delivery, have no bad taste such as smoke and alcohol, and the exclusion criteria comprise multiple deliveries, the age of the delivery person is less than 20 years old or more than 40 years old, diabetes, pre-pregnancy hypertension, chronic liver and kidney diseases, thyroid and other endocrine diseases, etc. The PE diagnosis standard refers to the guidelines for diagnosis and treatment of hypertensive disorders in gestational period (2015) published by the pregnant hypertensive disorders group of the obstetrics and gynecology department of the Chinese medical society. The project is approved by the general hospital ethics committee of Ningxia medical university, and the puerpera and the family members agree and sign an informed consent. Placenta tissues of patients are collected, and the height, the weight, the gestational period, the systolic pressure, the diastolic pressure, the birth weight of the newborn and other related clinical data of the two groups of pregnant women are collected. Within 10min after delivery of placenta, placenta tissue with umbilical cord as center in the central area of placenta is collected and kept away from calcification or bleeding focus. And (3) fixing the collected placenta tissues in 4% paraformaldehyde, and storing at-80 ℃ for later use, wherein the placenta tissues are used for preparing subsequent paraffin sections and detecting other indexes.
2.2 culture of human placental feeder cells
Culturing human extravillous trophoblasts (HTR-8/Svneo cells) in RPMI1640 medium containing 7% fetal bovine serum at 37 deg.C and 5% CO 2 Culturing in a cell culture box, allowing cells to grow in an adherent state, and carrying out passage for 1 time for 2-3 days.
2.3 Placental tissue Masson (Masson) staining
Recovering placenta tissue slice to room temperature, respectively soaking in xylene I for 15min and xylene II for 5min, gradient alcohol dehydration (100%, 95%, 85%, 75%) for 3min each time, and washing with distilled water for 3 times, 2min each time; dyeing Bouin liquid coal, incubating for 1h in an oven (water is added into a box to avoid dry slices) at 56 ℃; staining nuclei with hematoxylin solution for 20min, and washing with running water for 5min (tap water); acid reddening dye solution of ponceau rubra for 10min; washing with distilled water for 3 times (each time for 2min), dyeing with 1% phosphomolybdic acid for 7min, and directly dyeing with aniline blue for 15min without washing; soaking in 1% glacial acetic acid water solution for 1min, and washing with running water; 95% alcohol, 100% alcohol, xylene transparent, neutral gum sealing, and observing placenta tissue structure change with common microscope.
2.4 Immunoblotting for detecting MMP-2 (matrix metalloproteinase 2) expression change
Taking placenta tissue and HTR-8/SVneo cells (human chorionic trophoblast cells), preparing a protein lysate by PMSF (phenylmethylsulfonyl fluoride) and NP-40 (ethylphenyl polyethylene glycol) lysate according to a ratio of 1. Protein samples (300 μ g per well) were subjected to SDS-PAGE (sodium dodecyl sulfate-polyacrylamide gel) electrophoresis, the gel was transferred to 0.22 μm PVDF (polyvinylidene fluoride) membrane, 5% skim milk was blocked for 2 h at room temperature, PBST was washed for 10min for 3 times, β -actin antibody (1. The ratio of the optical density values of MMP-2 (matrix metalloproteinase 2) and beta-actin (beta-actin) is taken as the relative amount of MMP-2 (matrix metalloproteinase 2) protein expression.
2.4 RT-qPCR (real-time quantitative fluorescence PCR) for detecting lncRNA DINO (injury-induced non-coding RNA) expression
Total RNA (ribonucleic acid) from human placental tissue and HTR-8/SVneo cells (human chorionic trophoblast cells) was extracted according to the procedures of the Takara RNA kit and reverse transcribed according to the procedures of the reverse transcription kit. RT-qPCR (Real-Time quantitative fluorescent PCR) the expression level of lncRNA DINO (injury-induced non-coding RNA) was detected and analyzed by a Jena PCR instrument using Takara Real-Time PCR Master Mix (Green fluorescent dye-labeled Real-Time PCR mixture) kit. Beta-actin was used as a control. The experimental results were calculated according to the following formula: detection of relative expression level of target Gene =2 －△△Ct Wherein Δ Δ Ct = [ CtGI (detection sample) ]ctgapdh (detection sample)]- [ CtGI (calibration sample) -CtGAPDH (calibration sample)]. GI refers to the measured target gene, ct refers to the detected fluorescence signal intensity in the reaction system, and the calibration samples refer to all samples selected to represent 1 time of the expression level of the measured target gene.
2.5 Transfection of lncRNA DINO (Damage-induced non-coding RNA) interference fragment
Cell transfection was divided into 3 groups: blank control group (group 0), negative control transfection group (siNC group), DINO siRNA group (injury-induced interference fragment of non-coding RNA) (siDINO group). siNC (negative control transfection group) group: transfecting si RNA of a negative control; lncRNA DINO siRNA (injury-induced interference fragment of non-coding RNA) group: siRNA interfering fragments transfected with DINO (lesion-induced non-coding RNA). Respectively adding 5 μ l of negative control siNC and lncRNA DINO siRNA (injury-induced interference fragment of non-coding RNA) into 250 μ l of serum-free 1640 medium, gently mixing, mixing 6 μ l of Lipofectamine TM 2000 liposome and 250 μ l of serum-free medium, and standing at room temperature for 5min; mixing the two solutions, and standing at room temperature for 20min. Taking out the six-hole plate from the incubator, discarding the culture medium, adding the mixed solution into the corresponding holes respectively, metering the volume to 2 ml/hole by using the pure culture medium, putting the 6-hole plate into a constant-temperature incubator, and replacing the complete culture medium after 6 hours of transfection.
2.6 Cell scratching test for detecting migration capacity of trophoblast
A new 6-well plate was removed and two parallel lines were drawn at its bottom with a marker pen. Taking cells in logarithmic growth phase, according to 2X 10 5 Inoculating into 6-well plate at density of one cell/l, scribing two parallel scratches perpendicular to six-well plate with 200 μ l gun head when cell fusion degree reaches 90%, washing off detached cells with PBS, adding 2ml basal medium into each well, taking pictures under inverted microscope, determining scratch width as 0h migration distance, placing at 37 deg.C and 5% CO respectively 2 The constant-temperature incubator and the anoxic incubator continue to culture for 24h, the migration condition is observed under a microscope, the picture is taken, the migration distance of the 24h cells is obtained, and the experiment is repeated for 3 times to obtain an average value.
1.3.8 cell invasion assay trophoblast invasion alterations
Trypsinizing the cells to adjust the cell density to 1X 10 5 Cell/ml, matrigel (Matrigel) coated Transwell chamber basement membrane, upper layer cells diluted in fetal bovine serum free medium, transwell plate lower chamber added 7% fetal bovine serum containing RPMI1640 culture medium, cell culture plate placed in incubator 37 ℃,5% CO 2 After 24h of culture, the non-migrated cells on the upper surface were wiped clean with a cotton swab, fixed with 4% paraformaldehyde, stained with 0.1% crystal violet, incubated at 37 ℃ for 30min, cells were washed with PBS, the area of the image was randomly selected, and the experiment was repeated 3 times.
3. Statistical treatment
Statistical analysis was performed using GraphPad Prism 7.0 (biomedical mapping software), each bar graph being represented as mean ± Standard Deviation (SD). In selected experiments, one-way analysis of variance was used as appropriate, followed by Student-Newman-Kaul's test (Newman-Kelly's test: for comparison between groups) or unpaired Student's test (unpaired t test: for comparison between groups). p values less than or equal to 0.05 are considered statistically significant.
4. Results
4.1 Patient clinical data collection
Clinical data of patients are collected, and the analysis finds that: the difference between the clinical characteristics of height, weight, BMI, gestational period, age, etc. of the two groups of patients is not statistically significant (P > 0.05), while the difference between the clinical characteristics of systolic pressure, diastolic pressure, urinary protein, etc. is statistically significant (see FIG. 1).
4.2 Pre-eclampsia Patients (PE) and Normal pregnant women (PC) placental tissue architecture alterations
The placenta histology was observed by Masson (Masson) staining and the results showed: compared with the NC group, significant fibrin-like necrosis, myxoid changes, reduction of chorionic villi, and a large number of erythrocytes in the intervillous space were observed in the PE group (see fig. 2).
4.3 bioinformatics analysis of the Structure and position of Long non-coding RNAs (lncRNA DINO)
The position and structure information of DINO was analyzed using bioinformatics software (http:// asia. Ensemblel. Org /), showing: DINO is a LncRNA transcribed from CDKN1A, which is located at the position of chromosome 6 antisense strand 36677609-36678559, and this gene has only 1 transcript (splicing variation) with 1 exon, and is associated with 204 variant alleles, and the full length is 951bp. (see FIG. 3).
4.4 placental tissue DINO (Damage-induced non-coding RNA) protein expression
In order to clarify that the expression of DINO (injury-induced non-coding RNA) is changed in the placenta tissue, RT-qPCR is used for detecting the expression of LncRNA DINO (injury-induced non-coding RNA) in placenta trophoblasts of preeclamptic patients, and compared with a normal placenta group (NC group), the expression of LncRNA DINO (injury-induced non-coding RNA) in placenta tissue trophoblasts of the preeclamptic group (PE group) is increased and has significance (see figure 4).
4.5 detection of IncRNA DINO expression after transfection of interfering fragments
At the cellular level, after transfection of the interfering fragment si-DINO (interfering fragment of injury-induced non-coding RNA), RT-qPCR (real-time quantitative fluorescent PCR) detects the expression of lncRNA DINO (injury-induced non-coding RNA), and the results show that: the expression of lncRNA DINO (lesion-induced non-coding RNA) was significantly reduced (P < 0.05) compared to si-NC (negative control for interfering fragments) (see fig. 5).
4.6 modification of trophoblast MMP-2 (matrix Metalloproteinase 2) mRNA (messenger Single stranded ribonucleic acid) following interference with LncRNA DINO
The LncRNA DINO (damage-induced non-coding RNA) is transfected by the trophoblasts, the MMP-2 (matrix metalloproteinase 2) mRNA (messenger single-stranded ribonucleic acid) change is detected by RT-qPCR (real-time fluorescent quantitative PCR), and compared with the control group, the level of the DINO (damage-induced non-coding RNA) mRNA (messenger single-stranded ribonucleic acid) in the siDINO (damage-induced non-coding RNA) group is obviously increased compared with the control group, and the difference has statistical significance (see fig. 6).
4.7 altered protein expression of trophoblast MMP-2 (matrix Metalloproteinase 2) following interference with LncRNALnNCRNA DINO
The trophoblasts are transfected with LncRNAlencRNA DINO (injury-induced non-coding RNA), western blot detects that the protein expression of MMP-2 (matrix metalloproteinase 2) is changed, and compared with a control group, the protein level of the DINO (injury-induced non-coding RNA) in a siDINO (injury-induced non-coding RNA) group is obviously increased compared with that of the control group, and the difference has statistical significance (see figure 7).
4.8 interference of LncRNA DINO (Damage-induced non-coding RNA), scratch test for cell migration and invasion test for cell invasion changes
As expected, after interfering lncRNA DINO, the scratch test measures the cell migration and invasion ability, and siDINO (injury-induced non-coding RNA interference fragment) group has increased cell migration and invasion ability compared with the control group, and DINO (injury-induced non-coding RNA) can inhibit the migration and invasion of trophoblasts (see fig. 8).
5. Conclusion
The invention aims to provide a long-chain non-coding RNA marker for preeclampsia clinical risk assessment, the sample is obtained from placenta, the sample is convenient to obtain, no wound is caused to a patient, the long-chain non-coding RNA marker is generally accepted by the patient and family members of the patient, and the long-chain non-coding RNA marker has high feasibility.
The dysfunction of the invasion and migration of the trophoblast is an initiation link for regulating the occurrence and the development of preeclampsia and is also an important reason for the occurrence of the disease. Because of the specificity of pregnant women, the disease cannot be treated completely and effectively, and at present, clinical symptoms gradually disappear only after the fetus and the appendages (placenta) are delivered. DINO (injury-induced non-coding RNA) is used as a key target for regulating and controlling the invasion and migration of trophoblasts in placenta, can obviously improve the invasion and migration change of the trophoblasts, can be used as a long-chain non-coding RNA marker for preeclampsia clinical risk assessment, and has better feasibility and popularization.
The above description is only a preferred embodiment of the present invention, and the scope of the present invention is not limited thereto, and any simple modifications or equivalent substitutions of the technical solutions that can be obviously obtained by those skilled in the art within the technical scope of the present invention are within the scope of the present invention.
Sequence listing
<110> Ningxia medical university
<120> long-chain non-coding RNA for preeclampsia clinical risk assessment and application thereof
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acaaatctgg ctttttttac ttggagaatg agttggcact ctccaggagg acacagcact 780
gttagaatga gccccctttc tggctcaccg ctgacccact ctggcaggca aggatttacc 840
caatgcagct gaaaagatca ggaggatgac attaatacat aaaaattcat aaattataaa 900
aacgatgcac ctctctgcaa tttccagaaa agccccacaa tatcacctct attcccactg 960
atccctcact aggtcacctc tcccagaagc acctggagca cctagacacc ccaacaaagc 1020
atcttgaggc cagaatgatt tttcagtccc gtttatttca cagatgagga aattgaggtc 1080
cactgaactt aagtatacaa agttgttgat tgtcacatgc ttccgggaag gagggaattg 1140
gagagactac caaaaaaggg caacctgatc tccagggaaa cagaagaatt ggacattgaa 1200
ccaatctccc tacaccctac actcacctga acagaagaaa tccctgtggt tgcagcagct 1260
ttgttggcca ggaaggggag gatttgacga gtgagttgtc tgtctcctga atactcccca 1320
catagcccgt atacactgct ggggaaactg gggctcagag aagtctggtg acctaactca 1380
gatcatgcag caaagaaatg actatagttg gaacacagga cttttgcctc ctgcccgggg 1440
ctctctgctt gtcatccttt atttctgtgg ctccaaaatg acaaaaatgc caaataaccc 1500
tcatttgcag atggtttatg gagatgacat aaataaagga caattctgga agtgtctact 1560
ggttcttctg acagacacag aaatgagtga tgtgtctatc cgctcccatc tacctcacac 1620
ccctgacctc ccctggactt cacctttgcc tcctttctgt gcctgaaaca tttgcagttt 1680
tgcttttaaa aaattgcaga ggatggattg ttcatctgaa cagaaatccc actaaaaaac 1740
agaacccagg cttggagcag ctacaattac tgacatctca ggctgctcag agtctggaaa 1800
tctctgccca gacatgccca gtcttcttcc tctaacgcag ctgacctcgg ggaggacagg 1860
cttctttctc ctctgctgtg gggatgggag cccaggtagg ggtgggagga cacagtagca 1920
gacccccttg gcctgcctcg ccctggagtc aggccaggat tgtggctaaa ccccagaaag 1980
gccacggcac agcctcagga ccccactcta agccccactg ctgtctgcac cttcgctcct 2040
atacatccaa accatccaaa gggctggttg tcaaatgtcc agcagaggac aggcaccttc 2100
tcccactgcc ttgaagcccc tctgctttca ggcatttcaa atagactaga c 2151
<210> 2
<211> 16
<212> DNA/RNA
<213> primer sequence: upstream primer (lncRNA DINO)
<400> 2
ggtgcgctgg acacat 16
<210> 3
<211> 22
<212> DNA/RNA
<213> primer sequence: downstream primer (lncRNA DINO)
<400> 3
gtaaatcctt gcctgccaga gt 22
Claims (1)
1. The application of a reagent for detecting damage-induced non-coding RNA in preparing a product for diagnosing preeclampsia is characterized in that the nucleotide sequence of the damage-induced non-coding RNA is as follows: SEQ ID NO. 1.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN106480037A (en) * | 2016-12-20 | 2017-03-08 | 江苏省人民医院 | A kind of long non-coding RNA and the application in diagnosis preeclampsia and target drug treatment is prepared |
| CN109750038A (en) * | 2018-12-29 | 2019-05-14 | 烟台毓璜顶医院 | A kind of long non-coding RNA and the application in preparation diagnosis preeclampsia and target drug treatment |
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|---|---|---|---|---|
| CN106480037A (en) * | 2016-12-20 | 2017-03-08 | 江苏省人民医院 | A kind of long non-coding RNA and the application in diagnosis preeclampsia and target drug treatment is prepared |
| CN109750038A (en) * | 2018-12-29 | 2019-05-14 | 烟台毓璜顶医院 | A kind of long non-coding RNA and the application in preparation diagnosis preeclampsia and target drug treatment |
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| Homo sapiens cyclin dependent kinase inhibitor 1A (CDKN1A), RefSeqGene on chromosome 6;Kim KM;《NCBI》;20201005;第1-2页，ORIGIN * |
| lncRNA-SNHG5在重度子痫前期胎盘组织中的表达及其对滋养细胞增殖能力影响的机制;彭迎春等;《广西医学》;20200930(第18期);全文 * |
| 妊娠及胎盘疾病相关lncRNA的研究进展;唐昌青等;《实用妇产科杂志》;20180215(第02期);全文 * |
| 子痫前期胎盘组织miRNA差异表达谱生物学分析及miR-365a-3p参与发病的机制探讨;李晖等;《山东医药》;20200415(第11期);全文 * |
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