CN108977443A - A kind of movable circular rna of expression atherosclerosis and its application - Google Patents
A kind of movable circular rna of expression atherosclerosis and its application Download PDFInfo
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Abstract
The present invention relates to a kind of movable circular rna of expression atherosclerosis and its applications, it is characterized in that the circular rna is hsa_circ_0007478, and its nucleotide sequence is as shown in SEQ ID NO.1, expression of the application hsa_circ_0007478 in human atherosclerosis and its research of the adjustment effect to atherosclerosis biological function, provide the purposes of base hsa_circ_0007478 gene and its expression product in terms of the diagnosing and treating of atherosclerosis.
Description
Technical field
The present invention relates to a kind of circular rnas, and in particular to a kind of purposes of ring-type hsa_circ_0007478.
Background technique
Atherosclerosis (atherosclerosis, As) and its caused coronary atherosclerotic heart disease (coronary disease
Disease) it is cardiovascular most common disease, it is one of the main reason for mankind are lethal.Atherosclerosis is that coronary heart disease is formed and sent out
The main pathologic process of exhibition, main source of the macrophage as main composition and inflammatory factor in plaque,
Key effect has been played in the progression of atherosclerotic plaque.Macrophage energy synthesis and secretion various kinds of cell because
Son, and the stimulating expression of macrophage of these cell factors further secretes multiple inflammatory factors, lasting inflammatory conditions exacerbate
The unstability of patch, rupture easily occurs for unstable Coronary Atherosclerotic Plaque or ulceration, surface thrombosis are to make
At the main mechanism of acute coronary syndrome (including unstable angina, acute myocardial infarction AMI, sudden death), and acute coronary
Syndrome is then that coronary heart disease is lethal, the main reason for disabling.
In human genome, about 98% transcription not coding protein, and this non-coding RNA is in cell
Important role is play in growth, differentiation and human diseases.Wherein, circRNA is that one kind can be adjusted in post-transcriptional level
One group of non-protein coding RNA molecule of gene mRNA expression, it has the ring structure of closure, has a characteristic that 1, ring-type
RNA is more stable, and closed circular structure is not easy to be degraded by exonuclease RNaseR；2, highly conserved between kind；3, it is formed
Mode is mainly exon cyclisation；4, the wide expression in human body cell, even more than as many as 10 of its linear isomers times；
5, it is mostly present in the cytoplasm of eukaryocyte, there is tissue, timing and disease specific；6, there is the absorption of miRNA sponge
Function may act as competition endogenous endogenous RNA, in conjunction with miRNA, thus release miRNA to the inhibiting effect of its target gene,
Raise the expression of target gene.In recent years, with the gene expression regulation to circRNA in bion growth course
The understanding of effect so that the research of circRNA receives much attention, and becomes the hot spot of current life scientific theory and application study.
Therefore the variation and effect of circRNAs facilitate us into one during studying atherosclerotic course inflammatory activity
The pathophysiological process of the understanding atherosclerosis of step, for our clinical preventions and treatment atherosclerotic heart disease
New thinking is provided, also provides new action target spot for novel drug development and application.However there is which circRNAs on earth
Participate in not knowing during atherosclerosis is movable, and the function exercised of these circRNAs there are no from
Understand.
Summary of the invention
The object of the present invention is to provide to expression of the hsa_circ_0007478 in human atherosclerosis and its to dynamic
The research of the adjustment effect of pulse atherosclerosis biological function provides base hsa_circ_0007478 gene and its expression product
Purposes in terms of the diagnosing and treating of atherosclerosis.
The present invention provides one kind can express the movable circular rna of atherosclerosis, which is characterized in that described
Circular rna be hsa_circ_0007478, and its nucleotide sequence is as shown in SEQ ID NO.1.
The present invention provides a kind of atherosclerosis diagnosis kits, which is characterized in that the kit includes can
The primer of ring-type hsa_circ_0007478 as described in claim 1 is expanded, the amplimer is by such as SEQ ID NO:
The primer pair of the composition of DNA sequence dna shown in 7 and SEQ ID NO:8.
The present invention provides a kind of hsa_circ_0007478 gene as target gene in preparation treatment atherosclerosis
Drug in purposes, which is characterized in that the nucleotide sequence of the hsa_circ_0007478 gene such as SEQ ID NO.1
It is shown.
The present invention provides a kind of siRNA for inhibiting hsa_circ_0007478 gene expression, which is characterized in that described
SiRNA nucleotide sequence is as shown in SEQ ID NO.16 and can be used in inhibiting hsa_circ_0007478.
The present invention provides a kind of interference medicaments for atherosclerosis, which is characterized in that the interference medicament
Including such as above-mentioned siRNA.
Above-mentioned experimental result in order to obtain, itself uses following experimental program:
The screening technique of the movable correlation circRNAs of atherosclerosis a kind of and its verifying, specifically according to following step
It is rapid to implement:
The Macrophage Model of the outer ox-LDL stimulation of step 1, construct, using Arraystar Human circRNA core
Piece filters out the circRNAs of differential expression during the macrophage that ox-LDL is stimulated；
The Macrophage Model of the outer ox-LDL stimulation of step 2, construct, using the result of real-time quantitative PCR proofing chip.
The Macrophage Model of the outer ox-LDL stimulation of step 3, construct, interferes circRNA (hsa_ using siRNA
Circ_0007478), it was demonstrated that hsa_circ_0007478 can mitigate the inflammatory reaction of the macrophage through ox-LDL stimulation.
The features of the present invention also characterized in that
The Macrophage Model of the outer ox-LDL stimulation of step 1 construct, the pathology of simulation atherosclerosis reaction
Physiology course goes out difference during the macrophage of ox-LDL stimulation using Arraystar Human circRNA cDNA microarray
The specific steps of the circRNAs of expression are as follows:
Step 1.1, ox-LDL stimulation Macrophage Model building: be divided into two groups: macrophage group (control group) and
The macrophage group of ox-LDL stimulation.By person monocytic cell strain THP-1,48 hours formation macrophages are induced through PMA, are collected
The macrophage normally cultivated and through the post-stimulatory macrophage of ox-LDL, establishes the Macrophage Model of ox-LDL stimulation,
Cellular control unit is that person monocytic cell strain THP-1 is induced 43 hours formation macrophages through PMA；
1ml TRIZOL reagent lytic cell is added in step 1.2 in culture plate, repeatedly with rifle piping and druming more than 10 when cracking
It is secondary；0.2ml chloroform is added in TRIZOL, up and down oscillation 10 seconds, stands 5 minutes at room temperature, then at 4 DEG C, 12000rpm
High speed centrifugation 15 minutes, it is careful draw upper strata aqueous phase move into it is new without in RNA enzyme centrifuge tube；It is equal that upper water among the above is added
The isopropanol of volume after mixing, is stood overnight under the conditions of -20 DEG C, and then 12000rpm is centrifuged 10 minutes at 4 DEG C, is abandoned supernatant and is obtained
Precipitating；The ethyl alcohol 1ml that mass fraction is 75% is added in above-mentioned precipitating, overturns repeatedly for several times, 12000rpm centrifugation 5 at 4 DEG C
Minute, abandon supernatant；12000rpm is centrifuged 2 minutes at 4 DEG C again；Supernatant is abandoned, dries 1~2 minute at room temperature, adds 10 μ l without RNA
The dissolution of enzyme water, obtains RNA extracting solution, -80 DEG C save backup；Above-mentioned two groups of cell total rnas are extracted, it is fast with Agilent company
Fast Amp labelling kit labeled RNA, hybridizes with the probe on chip；
Step 1.3, the data that chip is collected using Agilent Scanner G2505C software, use Agilent
Feature Extraction software (version 11.0.1.1) software analysis data filters out variation multiple and is greater than
1.5 times, the circRNA of differential expression of the P value less than 0.05 (shown in Fig. 1)；
Step 1.4, using GO analysis and Pathway analysis pick out with atherosclerosis active procedure have it is close
Cut the circRNAs of relationship, i.e. hsa_circ_0007478, hsa_circ_0007085, hsa_circ_0092327 and hsa_
Circ_0082139, nucleotide sequence is respectively as shown in SEQ ID NO.1-SEQ ID NO.4.
The Macrophage Model of the outer ox-LDL stimulation of step 2 construct, using real-time quantitative PCR proofing chip as a result,
Specific step is as follows:
Step 2.1, experimental group are same as above: by person monocytic cell strain THP-1,48 hours formation macrophages are induced through PMA,
It collects the macrophage normally cultivated and through the post-stimulatory macrophage of ox-LDL, establishes the macrophage mould of ox-LDL stimulation
Type, cellular control unit are that person monocytic cell strain THP-1 is induced 48 hours formation macrophages through PMA；
The cracking of 1ml TRIZOL reagent will be added in step 2.2 in treated in step 2.1 cell, using above-mentioned step
Rapid 1.2 obtain RNA extracting solution, and -80 DEG C save backup；Reverse transcription reaction is carried out to RNA extracting solution with reverse transcription reagents to obtain
cDNA；
Step 2.3 carries out pcr amplification reaction to the cDNA solution in step 2.2, then is detected and expanded with fluorescence quantitative PCR instrument
Increase result, the results showed that the circRNAs with atherosclerosis process close relation, respectively hsa_circ_
0007478, hsa_circ_0007085, hsa_circ_0092327 and hsa_circ_0082139, using GAPDH as internal reference (figure
Shown in 2).
The primer of the hsa_circ_0007478 are as follows: F:5 ' ATACATCTGCTGCCTACATTCC3, R:5 '
AGTCCCCAAGTACCAAGTGC3'；The primer of the hsa_circ_0007085 are as follows: F:5 '
CTCCATCAGGAAAACAAGCC3 ', R:5 ' CGTAGGTCCCTGAGAAAATGTT3 '；The hsa_circ_0092327's draws
Object are as follows: F:5 ' GGCATTAGAAAGCCAGGGAC3 ', R:5 ' ACTGACCAGATAGCCATCACC3 '；The hsa_circ_
0082139 primer are as follows: F:5 ' CCACTGTCACCATTGGAGGA3 ', R:5 ' ACAGGGGACGCAGTTTCTTG3 ', it is described
The primer of GAPDH are as follows: F:5 ' GGGAAACTGTGGCGTGAT3 ', R:5 ' GAGTGGGTGTCGCTGTTGA3 '.
The Macrophage Model of the outer ox-LDL stimulation of step 3, construct, interferes circRNA (hsa_ using siRNA
Circ_0007478), it was demonstrated that hsa_circ_0007478 can mitigate the inflammatory reaction of the macrophage through ox-LDL stimulation,
This provides a kind of new thinking for the pathophysiological process of our studying atherosclerotic inflammatory reactions, provides for coronary heart disease
New molecule foundation or therapy target.
Second technical solution of the present invention is the relevant hsa_circ_ of atherosclerosis activity
0007478 in the application for predicting and preventing and treating myocardial infarction, and discovery is opposite with coronary angiography patients with negative, hsa_circ_
0007478 expresses in acute myocardial infarction patient blood in high, it was confirmed that hsa_circ_0007478 and coronary heart disease
The correlation of acute myocardial infarction AMI development, provides new target molecule for the early diagnosis and biological therapy of myocardial infarction.
The beneficial effects of the present invention are: present invention demonstrates 4 circRNAs in atherosclerosis reaction process
Expression quantity changed, the analysis by bioinformatics finds that wherein 1 hsa_circ_0007478 and artery are athero-
Hardening inflammatory reaction plays the role of close relationship and can play inhibition inflammatory reaction after lowering the circRNA, and
Compared with coronary angiography feminine gender group, in high expression, this is hsa_circ_0007478 level in coronary heart disease acute myocardial infarction group blood
The pathophysiological process of our studying atherosclerotics provides a kind of new thinking, and mentions for the prediction and prevention and treatment of coronary heart disease
For new molecule foundation or therapy target.
Detailed description of the invention
Fig. 1 is the chip hybridization detection figure (schematic diagram of circRNA chip hybridization) of two groups of cell samples of embodiment 1.
Fig. 2 is the real-time quantitative PCR proof diagram of 4 circRNAs of Macrophage Model of the ox-LDL stimulation of embodiment 2
(each circRNAs relative expression quantity schematic diagram).
Fig. 3 is Western blot schematic diagram (each group IL-1 β relative expression quantity signal of the group cell sample of embodiment 3
Figure).
Fig. 4 is Western blot schematic diagram (each group NLRP3, p-P65 and the P65 phase of the group cell sample of embodiment 3
To expression quantity schematic diagram).
Fig. 5 is the hsa_circ_0007478 of embodiment 4 in coronary angiography feminine gender and coronary heart disease acute myocardial infarction patient blood
Middle expression quantity schematic diagram.
Specific embodiment
The following describes the present invention in detail with reference to the accompanying drawings and specific embodiments.
Firstly, the Macrophage Model of the outer ox-LDL stimulation of construct, using Human circRNA Array (8x15K,
Arraystar) cDNA microarray goes out the circRNAs step of differential expression in the Macrophage Inflamatory reaction process of ox-LDL stimulation
It is as follows:
A. experiment is divided to two groups: control group and the post-stimulatory macrophage of cx-LDL are by person monocytic cell strain THP-1, through PMA
48 hours formation macrophages are induced, the macrophage normally cultivated is collected and through the post-stimulatory macrophage of ox-LDL, are built
The Macrophage Model of vertical ox-LDL stimulation, cellular control unit are that person monocytic cell strain THP-1 is induced 48 hours shapes through PMA
At macrophage.
B. it cultivates the homogenate of cell: the cell of 1ml TRIZOL cracking culture being added directly in culture dish, when cracking is used
Rifle suction is beaten blows and beats more than 10 times repeatedly.
C. 0.2ml chloroform is added in the TRIZOL of step b, up and down oscillation 10 seconds, at standing 5 minutes, 4 DEG C at room temperature,
12000rpm high speed centrifugation 15 minutes, draw upper strata aqueous phase move into it is new without in RNA enzyme centrifuge tube；
D, isopropanol precipitating: be added with the step c isometric isopropanol of water phase at the middle and upper levels, it is quiet under the conditions of -20 DEG C after mixing
It sets overnight, then 12000rpm is centrifuged 10 minutes at 4 DEG C, and abandoning supernatant must precipitate；
E, ethanol washing: the ethyl alcohol 1ml that mass fraction is 75% is added in the precipitating of step d, overturns for several times, at 4 DEG C
12000rpm is centrifuged 5 minutes, abandons supernatant；
F, dry RNA: after previous step abandons supernatant, 12000rpm is centrifuged 2 minutes at 4 DEG C again；Supernatant is abandoned, is done at room temperature
Dry 1~2 minute, adds 20 μ l to dissolve without RNA enzyme water, obtain RNA extracting solution, -80 DEG C save backup；
G. above-mentioned two groups of cell total rnas are extracted, with the quick Amp labelling kit labeled RNA of Agilent company, as a result
It is shown in Table 1:A1, A2, A3 (repetition of biology three) and represents Normal group, B1, B2, B3 (repetition of biology three) represent ox-LDL
The RNA of the macrophage group of stimulation, above-mentioned each group hybridizes with the probe on chip, and as a result A, B, C figure indicate just as shown in Figure 1:
With the schematic diagram of chip hybridization, D, E, F scheme to indicate that the macrophage group RNA of ox-LDL stimulation is miscellaneous with chip the cell RNA often organized
The schematic diagram of friendship.
H. the data for collecting chip, with Agilent Feature Extraction software (version
11.0.1.1) software analysis data filters out variation multiple and is greater than 1.5 times, differential expression of the P value less than 0.05
circRNAs。
The RNA information table of two groups of 1 embodiment 1 of table and gene chip hybridization
I. pick out may to the relevant circRNAs of atherosclerosis activity (hsa_circ_0007478,
Hsa_circ_0007085, hsa_circ_0092327 and hsa_circ_0082139) (its nucleotide sequence such as SEQ ID
NO.1-SEQ ID NO.4) it is tested to subsequent authentication.
The macrophage of above-described ox-LDL stimulation can simulate clinically coronary heart disease cardiovascular artery congee well
The pathologic process of sample hardening inflammatory reaction.Furthermore from table 1 it follows that rna stability is high in above-mentioned RNA extraction method
It is not degradable, the favorable repeatability of experiment, therefore the result credibility of circNRA cDNA microarray is high.
Secondly, the Macrophage Model of the outer ox-LDL stimulation of construct, using the result of real-time quantitative PCR proofing chip.
A. the Macrophage Model of the outer ox-LDL stimulation of construct, with described in a in embodiment 1.
B. cultivate the homogenate of cell: by step a through 1ml TRIZOL examination is added in ox-LDL treated foam cells
Agent cracking beats more than 10 times with rifle suction repeatedly when cracking, obtains RNA extracting solution according still further to the c-f step operation in embodiment 1 ,-
80 DEG C save backup；
C, PCR is detected: carrying out pcr amplification reaction to the cDNA solution of step g, amplification upstream and downstream primer is circRNA special
Specific amplification upstream and downstream primer or GAPDH amplification upstream and downstream primer (as shown in table 2) (SEQ ID NO.5-NO.14), then with glimmering
The detection of Fluorescent Quantitative PCR instrument.It is as shown in Figure 2: in the Macrophage Model of ox-LDL stimulation, 3 circRNAs (hsa_circ_
0007478, hsa_circ_0007085, hsa_circ_0092327) it is raised compared with control group, 1 circRNA (hsa_circ_
0082139) it is lowered compared with control group, it is as a result consistent with chip results.Further confirm that this 4 circRNAs are pierced in ox-LDL
Changed in sharp Macrophage Inflamatory reaction process, pathophysiological process among these may be participated in, is subsequent
Functional study provides clue and thinking.
2 real-time quantitative PCR of table uses list of primers
Furthermore hsa_circ_0007478 is interfered using siRNA, after the Macrophage Model for observing external ox-LDL stimulation
The variation of inflammation index, using the effect of real-time quantitative PCR observation interference, WB detects IL-1 β, NLRP3, p-P65 and P65
Situation of change
A.siRNA interferes intracellular hsa_circ_0007478, picks out a most suitable siRNA for subsequent reality
It tests: small interference fragment (3) (its target sequence such as SEQ ID NO.15-SEQ ID of synthesis needle hsa_circ_0007478
NO.17), sequence 1: positive-sense strand: TTGTGCTTCCCTAGACTCT；Sequence 2: positive-sense strand: TTCCCTAGACTCTCCTTTC；Sequence
Column 3: positive-sense strand: CCTAGACTCTCCTTTCCAA；THP-1 cell is laid in 6 orifice plates and PMA is added to stimulate 48 hours, when thin
Born of the same parents' density reaches 60% and starts to transfect siRNA.Each transfection sample standard deviation is prepared in accordance with the following steps:
1) it dilutes siRNA: diluting 1.25 μ l, 20 μM of siRNA storages with 50 μ l 1 × riboFECTTM CP Buffer
Liquid mixes gently, and is incubated at room temperature 5 minutes.
2) prepared by mixed liquor: 5 μ l riboFECTTM CP Reagent is added, gently piping and druming mixes, and is incubated at room temperature 15 points
Clock.
3) riboFECTTM CP mixed liquor is added in 443.75 μ l cell culture mediums, is mixed gently.
4) culture plate is placed in normal oxygen incubator and is cultivated 48 hours, detected described in b to the h step in Application Example 2
Optimal one group of hsa_circ_0007478 jamming effectiveness.According to the interference of the result siRNA interference sequence 2 of real-time quantitative PCR
Effect is best, therefore is used for subsequent experiment.
B. the macrophage after interfering is after ox-LDL is stimulated with the effect of real-time quantitative PCR detection interference: experiment point
Are as follows: Normal group；Ox-LDL stimulation group；Ox-LDL+siRNA NC group；Ox-LDL+siRNA group；Wherein ox-LDL+siRNA
Control group of the sequence of NC group transfection as transfection experiment.After transfecting siRNA according to step described in a in embodiment 3, then press
Ox-LDL stimulation is carried out according to described in a in embodiment 1 pairs of three groups of cell another in addition to control group, the b in last Application Example 2
To the expression quantity for detecting hsa_circ_0007478 described in h step, whether verifying interference succeeds.
The table of the relevant index IL-1 β of inflammation after c.Western blot method detection interference hsa_circ_0007478
It reaches: transfecting cell according to the step a applied in example 3, this 4 groups of cells are handled according still further to described in a in embodiment 1, finally
Use Western blot method observing effect.As shown in Figure 4: ox-LDL stimulation group is obvious compared with the expression quantity of control group IL-1 β
Increase, the IL-1 β expression quantity of ox-LDL+siRNANC group compared with ox-LDL stimulation group without significant change, however ox-LDL+siRNA
IL-1 β expression quantity in group has apparent attenuating compared with ox-LDL stimulation group, it was demonstrated that interference circRNA (hsa_circ_0007478)
The inflammatory reaction of the macrophage through ox-LDL stimulation can be mitigated.
D.Western blot method detection interference circRNA (hsa_circ_0007478) afterwards NLRP3 inflammation corpusculum and
The expression of associated signal paths: transfecting cell according to the step a that applies in example 3, according still further to described in a in embodiment 1 to this 4 groups
Cell is handled, and Western blot method observing effect is finally used.As shown in Figure 4: ox-LDL stimulation group is compared with control group
The expression quantity of NLPR3, p-P65 and P65 obviously increase, the table of NLPR3, p-P65 and P65 of ox-LDL+siRNA NC group
Up to amount compared with ox-LDL stimulation group without significant change, however the expression quantity of NLPR3, p-P65 in ox-LDL+siRNA group are compared with ox-
LDL stimulation group has apparent reduction, it was demonstrated that it is thin that interference circRNA (hsa_circ_0007478) mitigates ox-LDL stimulation macrophage
The inflammatory reaction of born of the same parents' cell is by inhibiting the relevant NLRP3 inflammation corpusculum approach of P65 signal path to realize.
Finally, being detected in coronary angiography patients with negative and coronary heart disease acute myocardial infarction patient blood using real-time quantitative PCR
The expression quantity of hsa_circ_0007478, the specific steps are as follows:
A. the blood specimen of coronary angiography feminine gender and coronary atherosclerotic heart disease patients of acute myocardial infarction is collected:
Coronary angiography feminine gender inclusion criteria: narrow and electrocardiogram is had no in the parallel coronary angiography prompt coronary artery blood vessel of our hospital's hospitalization
And Myocardial Enzymologic/troponin no abnormality seen patient；Coronary atherosclerotic heart disease patients of acute myocardial infarction:
It raises in our hospital's hospitalization parallel coronary angiography prompt coronary artery hemadostewnosis/occlusion, electrocardiogram prompt ST, Myocardial Enzymologic/flesh
The raised patient of calcium albumen progressive；Exclusion criteria: hemodynamic instability person or patients with cardiogenic shock；Serious liver function
Energy failure, glutamic-pyruvic transaminase >=3 times or more person；Obvious autoimmune disease；The thyroid dysfunction not controlled；
Malignant tumour；The general infection of active stage；Pregnant woman, lactation person；Other clinical test persons are being participated in nearly three months；Spirit
Disease；Refusal participates in patient.The age of two groups of patients, gender are identical；The matching of hypertensive diabetes situation.
The venous blood 5ml of above-mentioned two groups of patients is collected, is deposited in -80 DEG C of refrigerators for use after extracting mononuclearcell.
B. quantitative expression of the real-time quantitative PCR detection hsa_circ_0007478 in mononuclearcell；
The cracking of 1ml TRIZOL reagent is added in sample after thawing, more than 10 times are beaten with rifle suction repeatedly when cracking, according still further to implementation
C-f step operation in example 1 obtains RNA extracting solution, and -80 DEG C save backup；Biorad company C1000 (Biosystems,
USA the reaction of SYBR Green quantitative fluorescent PCR) is carried out on instrument, as a result as shown in figure 5, hsa_circ_0007478 is in coronary disease
Sick acute myocardial infarction patient expresses with respect in the blood of coronary angiography patients with negative in high.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
For member, without departing from the technical principles of the invention, several improvements and modifications can also be made, above-mentioned hypothesis this
A little improvement and modification also should be regarded as protection scope of the present invention.
Claims (6)
1. one kind can express the movable circular rna of atherosclerosis, which is characterized in that the circular rna is
Hsa_circ_0007478, and its nucleotide sequence is as shown in SEQ ID NO.1.
2. a kind of atherosclerosis diagnosis kit, which is characterized in that the kit includes that can expand such as claim 1
The primer of the hsa_circ_0007478.
3. atherosclerosis diagnosis kit according to claim 2, which is characterized in that the amplimer is by such as
The primer pair of the composition of DNA sequence dna shown in SEQ IDNO:7 and SEQ ID NO:8.
4.hsa_circ_0007478 purposes of the gene as target gene in the drug of preparation treatment atherosclerosis, special
Sign is that the nucleotide sequence of the hsa_circ_0007478 gene is as shown in SEQ ID NO.1.
5. a kind of siRNA for inhibiting hsa_circ_0007478 gene expression, which is characterized in that the siRNA nucleotides sequence
Column are as shown in SEQ ID NO.16 and can be used in inhibiting hsa_circ_0007478.
6. a kind of interference medicament for atherosclerosis, which is characterized in that the interference medicament includes that right such as is wanted in 5
The siRNA.
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| CN111455037A (en) * | 2020-04-02 | 2020-07-28 | 广东医科大学 | Coronary heart disease molecular diagnosis marker based on plasma exosome cyclic RNA and application thereof |
| CN112646873A (en) * | 2020-12-29 | 2021-04-13 | 上海市中医老年医学研究所 | Reagent for detecting atherosclerosis and application thereof |
| CN115896101A (en) * | 2022-10-26 | 2023-04-04 | 江苏省人民医院（南京医科大学第一附属医院） | Protein translated by circular RNA molecule and application thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111455037A (en) * | 2020-04-02 | 2020-07-28 | 广东医科大学 | Coronary heart disease molecular diagnosis marker based on plasma exosome cyclic RNA and application thereof |
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| CN112646873A (en) * | 2020-12-29 | 2021-04-13 | 上海市中医老年医学研究所 | Reagent for detecting atherosclerosis and application thereof |
| CN112646873B (en) * | 2020-12-29 | 2021-08-03 | 上海市中医老年医学研究所 | Reagent for detecting atherosclerosis and application thereof |
| CN115896101A (en) * | 2022-10-26 | 2023-04-04 | 江苏省人民医院（南京医科大学第一附属医院） | Protein translated by circular RNA molecule and application thereof |
| CN115896101B (en) * | 2022-10-26 | 2023-12-05 | 江苏省人民医院（南京医科大学第一附属医院） | Protein translated by circular RNA molecule and application thereof |
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