CN105521490B - Applications of the miRNA-888-3p in Degenerative disc disease diagnosis and treatment - Google Patents
Applications of the miRNA-888-3p in Degenerative disc disease diagnosis and treatment Download PDFInfo
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- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
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Abstract
The present invention relates to applications of 888 3p of miRNA in Degenerative disc disease diagnosis and treatment.The experiment proved that 888 3p of miRNA can efficiently differentiate the sample of Degenerative disc disease sample and Healthy People.The invention also discloses the applications in the drug that 888 3p of miRNA prepare treatment Degenerative disc disease.The present invention provides new method for clinical diagnosis Degenerative disc disease on a molecular scale, while new target spot is provided for the gene therapy of Degenerative disc disease.
Description
Technical field
The invention belongs to biomedicine field, diagnosed in Degenerative disc disease more particularly to miRNA-888-3p,
Purposes in treatment.
Background technology
Microrna (miRNA or microRNA) is the endogenous non-coding that a kind of length is 18~25 nucleotide single-chains
Property tiny RNA, is to be generated under the processing of enzyme by precursor, the sequence of precursor is generally 70~120 nucleotide.By with target gene
Sequence-specific interaction adjusts gene expression in transcriptional level, participates in various biological process, and evolutionary process is conservative.It is right
The Systematic Analysis of miRNA space expressions shows that many miRNAs are expressed with tissue specific way.Each miRNA can be with
The complex for being positioned at a variety of mRNA, miRNA and RISC can be with target gene mRNA5 '-UTR or 3 '-by base pairing
Complementary series in UTR is combined, and inhibits protein translation, or causes mRNA degradations, thus the expression of negative regulation target gene,
Therefore a variety of transcriptons may be fine-tuned by miRNA, so that transcription-translation system is effectively operated, and then cell is able to
The rapidly and efficiently cell event of control threshold dependence.Generally believe that the function of miRNA is some for participating in life at present
Basic process such as growth and development, orga- nogenesis, hematopoiesis, Cell apoptosis and proliferation, stress reaction and tumour generation etc..
Degenerative disc disease refers to that cell-mediated biology occurs under many reasons comprehensive function for disc tissue
Chemical change causes aging acceleration, interverbebral disc mechanical characteristic to change, and makes to close on Bones and joints, ligament generation respective change, causes ridge
Column is unstable, and compressing spinal cord, nerve root, artery cause the syndrome of corresponding clinical symptoms and sign.It includes discogenic waist
Bitterly, the protrusion of the intervertebral disc, degeneration spinal instability disease, degenerative stenosis, degeneration Spondylolisthesis etc., are clinically common
One of disease.Its cause of disease is unclear, generally believes that Degenerative disc disease is that h and E factor is total at present
The result of same-action.With the development of technique of gene detection, in recent years, people gradually recognize genetic predisposition i.e. inherent cause
For disclosing the important function of Degenerative disc disease morbidity.
Degenerative disc disease generally takes expectant treatment at present, i.e., from expectant treatment successively to minimally invasive, routine operation
Treatment, non-fused fixation for treatment, the ladder for merging fixation for treatment.The treatment of advanced ladder, wound is bigger, to body nature
The intervention of anatomic status is bigger, and the treatment of each advanced ladder can be used as the remedial measure to relatively low level stepped care.To suffering from
The therapeutic scheme that person takes will be according to the stage residing for conditions of patients, while being accounted for individual patients factor, such as wound, wind
The wish of danger, expense and patient.But no matter which kind of modus operandi, cannot be carried out effectively early stage intervertebral disc degeneration
Intervene, cannot delay or the development of reverse disease.Open surgery (discectomy fusion internal fixation) though can more thoroughly
Degeneration intervertebral disc is cut off, but operation wound is big, costly, interior fixation limits the mobility of backbone, accelerates neighbouring section intervertebral
The regression of disk.Minimally Invasive Surgery cannot thoroughly cut off degeneration intervertebral disc tissue, postsurgery symptoms may not be able to complete incidence graph, postoperative recurrence
Possibility it is larger.
Gene therapy is that the treatment of intervertebral disc degeneration opened up a new way, and achieve inspirer achievement.At present
Through reporting some miRNA such as miRNA-623, miRNA-625 related with Degenerative disc disease, but current gene therapy
Research remain in the infrastest stage, if also having many problems urgently to be resolved hurrily applied to clinical.
Invention content
In order to make up for the deficiencies of the prior art, the purpose of the present invention is to provide a kind of miRNA-888-3p moves back in interverbebral disc
Application in row diagnosis of disease.
To achieve the goals above, present invention employs following technical solutions：
The present invention provides miRNA-888-3p to prepare the application in treating Degenerative disc disease drug, described
The nucleotide sequence of miRNA-888-3p is as shown in SEQ ID NO.1.The experiment proves that miRNA-888-3p and intervertebral
Disk degenerative disease is related,
Further, the drug includes miRNA-888-3p inhibitor.
Further, the miRNA-888-3p inhibitor can inhibit the expression of miRNA-888-3p or can inhibit
The function of miRNA-888-3p.
Further, miRNA-888-3p inhibitor includes albumen, oligonucleotides, micromolecular compound.
Preferably, the miRNA-888-3p inhibitor is the antisense oligonucleotides or antagonist of miRNA-888-3p.
The present invention provides a kind of drug for treating Degenerative disc disease, the drug includes above-mentioned miRNA-
888-3p inhibitor.
The specific antisense oligo for going out it according to miRNA-888-3p sequence designs, antisense oligonucleotides is transferred to
After in human body, they can obviously lower the expression of miRNA-888-3p." antisense oligonucleotides (antisense-
Oligonucleotides, AS-Ons or ASO) " be also known as " GEM 132 ", refer to length be about 18-26nt (more particularly
About 19-22nt) DNA molecular or RNA molecule or its analog.
In the present invention, " antisense oligonucleotides " further includes using as based on nucleic acid lock or nucleic acid chains backbone modification
The modified GEM 132 that the means such as technology obtain, the modification do not change the activity of antisense oligonucleotides substantially, more
Goodly, described to modify the stability, activity or therapeutic effect that antisense oligonucleotides can be improved.Nucleic acid locks (locked nucleic
Acid, LNA) typically refer to the modification skill for 2 ' oxygen atoms of ribose and 4 ' carbon atoms being connected by a methylene bridge
Art.In solubility, nuclease-resistant degradation etc. has greatly to be changed the antisense drug that modification technique based on nucleic acid chain backbone develops
It is kind, and be easy to largely synthesize.There are many backbone modification methods of oligonucleotides, including thio method, such as by deoxynucleotide chain
Thio-modification is thio deoxynucleotide chain.This method is to substitute the oxygen atom of the phosphate bond on DNA skeletons with sulphur atom, can
Resist nuclease degradation.It should be understood that any largely or entirely active modification that can keep the antisense oligonucleotides is all
Including in the present invention.
The drug of the treatment Degenerative disc disease of the present invention is also comprising pharmaceutically acceptable carrier, the load
Body includes but not limited to：Diluent, buffer, suspension, emulsion, granule, encapsulation agents, excipient, filler, adhesive,
Spray, cutaneous permeable agent, wetting agent, disintegrant, sorbefacient, surfactant, colorant, corrigent or absorption carry
Body.
Including but not limited to microinjection agent, the dosage form suitable for transfection, injection, tablet, powder can be made in the drug
Agent, granula, capsule.The drug of above-mentioned various dosage forms can be prepared according to the conventional method of pharmaceutical field.
The drug can be administered alone or the drug of Degenerative disc disease can be inhibited to be combined with other
Using.
The drug can be applied in vitro：By the antisense oligonucleotides of miRNA-888-3p or antagonist, miRNA-
The expression vector of 888-3p antisense oligonucleotides imports or transfects in vitro human body itself or variant cell (or heterogenous cell), warp
After vitro cell expansion, defeated the Huis' body.
The drug can be applied in vivo：By the antisense oligonucleotides of miRNA-888-3p or antagonist, miRNA-
The expression vector of 888-3p antisense oligonucleotides is introduced directly into vivo.This carrier can be virus type or non-viral, even
It is naked DNA or RNA.
The subject can be the mankind or other mammals.More specifically, subject be organ, it is tissue, thin
Born of the same parents.
The nucleic acid molecules of the present invention can be the form of RNA, DNA, PNA, LNA.
The present invention provides applications of the miRNA-888-3p in preparing Degenerative disc disease early diagnosis kit.
The present invention also provides a kind of kits of diagnosis early stage Degenerative disc disease, and the kit includes being used for
Detect the reagent of the expression of miRNA-888-3p.Compared with the expression of the miRNA-888-3p in Healthy People tissue,
If the expression for detecting miRNA-888-3p in subject's tissue by kit significantly increases, judge that the subject suffers from
Degenerative disc disease.
Further, above-mentioned reagent includes the primer and/or probe for miRNA-888-3p.The reagent further includes needle
To in the prior art it has been reported that can be used for diagnose Degenerative disc disease miRNA primer and/or probe.It will be a variety of
The detection primer and/or probe of miRNA is placed in same reagent box judges interverbebral disc by detecting a variety of miRNA indexs joints
The case where degenerative disease, is also contained within protection scope of the present invention.
The present invention provides a kind of chip of diagnosis Degenerative disc disease, the chip includes solid phase carrier and consolidates
The oligonucleotide probe being scheduled on the solid phase carrier, the oligonucleotide probe include specifically corresponding to miRNA-888-
Some or all of 3p sequences.The oligonucleotide probe may also include in the prior art it has been reported that can be used for diagnosing
The oligonucleotide probe of the miRNA of Degenerative disc disease leads to the detection probe placement of a variety of miRNA on the same chip
Cross detect the case where a variety of miRNA indexs joint judges Degenerative disc disease be also contained in protection scope of the present invention it
It is interior.
Further, the solid phase carrier includes the various common used materials that genetic chip field can be used in the solid phase carrier,
Such as, but not limited to nylon membrane, slide or silicon chip through active group (such as aldehyde radical, amino) modification, unmodified slide, modeling
Tablet etc..
The conventional manufacturing method of biochip known in the art can be used in the preparation of the miRNA chips, for example, such as
Fruit solid phase carrier is gone here and there using modification slide or silicon chip, 5 ' ends of probe containing amido modified poly- dT, can be by oligonucleotides
Probe is configured to solution, then uses point sample instrument that its point on modification slide or silicon chip, is arranged in scheduled sequence or array,
Then it is fixed by standing overnight, so that it may obtain the miRNA chips of the present invention.If nucleic acid is without amido modified, system
Preparation Method can also refer to：Wang Shenwu chief editors'《Gene diagnosis technology-on-radiation operation manual》；J.L.erisi,
V.R.Iyer, P.O.BROWN.Exploring the metabolic and genetic control of gene
Expression on a genomic scale.Science, 1997；278：680 and Ma Li people, Jiang Zhonghua edit biology cores
The Beijing piece：Chemical Industry Press, 2000,1-130.
The miRNA-888-3p of the present invention can be natural or artificial synthesized, or use can express miRNA-
The carrier transfectional cell of 888-3p obtains.The carrier includes viral vectors, non-viral carrier, eukaryotic vector.
Viral vectors can be any carrier appropriate, including but not limited to retroviral vector, adenovirus vector, gland
Viral related viral vectors, herpesviral (such as herpes simplex virus, vaccinia virus and Epstein-Barr virus) carrier, alphavirus vectors.
Non-virus carrier includes naked plasmid dna carrier, as pCMV-Myc expression vectors, pcDNA3.0 expression vectors,
PcDNA3.1 expression vectors, pEGFP expression vectors, pEF Bos expression vectors, pTet expression vectors, pTRE expression vectors or
Modified carrier, such as pBin438, pCAMBIA1301 etc. on the basis of known expression vector；Cationic polymer, it is main
To include polyethyleneimine, poly-L-lysine, arginine-rich protein, polyethylene glycol and cationic copolymerization body etc.；Based on anti-
The target gene transmission system Histones of body；Liposome or lipid complex are current most commonly used non-viral loads
Body.
It should be known that the miRNA-888-3p of the present invention includes the functional equivalent of composing type nucleic acid molecules, i.e. variant.
Those skilled in the art are known, in order to ensure the stability of miRNA, can increase in the one or both ends of miRNA and protect
Shield property base, such as TT, can also modify miRNA bases, but not influence the function of miRNA.Therefore, people in the art
Member is known, under conditions of not influencing miRNA-888-3p functions, carries out base modification to miRNA-888-3p or at both ends
Increase the sequence that base obtains to be also contained within protection scope of the present invention.
The advantages of the present invention：
Present invention firstly discovers that miRNA-888-3p expression is related to Degenerative disc disease, pass through detection subject
The expression of miRNA-888-3p in tissue, it can be determined that whether subject suffers from Degenerative disc disease, to instruct
Clinician provides prevention scheme or therapeutic scheme to subject.
Description of the drawings
Fig. 1 shows the expression in Degenerative disc disease tissue using QPCR detections miRNA-888-3p；
Fig. 2 shows the expression in Degenerative disc disease cell line using QPCR detections miRNA-888-3p；
Fig. 3 shows the inhibiting effect that anti-miRNA-888-3p expresses miRNA-888-3p.
Specific implementation mode
It being further illustrated the present invention with reference to specific embodiment, the embodiment of the present invention is only used for explaining the present invention,
It is not intended to limit protection scope of the present invention.
Experimental method used in following embodiments is conventional method unless otherwise specified.
The materials, reagents and the like used in the following examples is commercially available unless otherwise specified.
The screening of embodiment 1 and the relevant miRNA of Degenerative disc disease
1, sample acquisition：Respectively collect 10 Healthy People tissue samples and Degenerative disc disease patient's nucleus pulposus sample
This.Agreement of the acquirement of above-mentioned all samples by the committee of organizational ethics.
2, the extraction of sample total serum IgE
Shift to an earlier date total serum IgE using the tissue RNA extracts kits of QIAGEN companies.It is as follows：
1) in the clear area of less RNase interference, tissue samples about 20mg is weighed using the mortar containing appropriate liquid nitrogen, uses pestle
Stick is ground to powdered；
2) sample is transferred in the centrifuge tube of a 2ml without RNA enzyme；
3) 300 μ l Lysis solution are added, is placed in homogenizer, is fully ground 1-5min；
4) 12000g 4 DEG C, centrifuges 10min, shifts in supernatant to the centrifuge tube of new 1.5ml；
5) 600 μ l RNase-Free Water are added, with vortex device mixing；
6) 20 μ l Proteinase Ks are added, the warm bath 15min in 55 DEG C of water-baths, be constantly vortexed mixing；
7) 14000g, room temperature centrifuge 1min, make pellet cell debris in centrifugation bottom of the tube, supernatant is taken to be transferred to other one
In a centrifuge tube without RNA enzyme 1.5ml；
8) 95% ethyl alcohol of 450 μ l, vortex mixing is added；
9) 650 lysates of the μ l containing ethyl alcohol are taken to be added in centrifugal column, 14000g centrifuges 1min；Lower layer is abandoned, again by pillar
It is placed in collecting pipe；
10) according to the capacity of lysate, step 9) is repeated；
11) 400 μ l Wash solution, 14000g are added and centrifuge 2min；Lower layer is abandoned, column is placed in a new collecting pipe
In；
12) 100 μ l Enzyme Incubation Buffer are added and 15 μ l DNase I, 14000g centrifuge 1min, it will
Solution in collecting pipe moves into column again, is placed at room temperature for 15min；
13) 400 μ l Wash solution, 14000g centrifugation 1min are added, abandons lower layer, pillar is replaced in collecting pipe
In；
14) 400 μ l Wash solution, 14000g centrifugation 2min are added, abandons collecting pipe, pillar is put into 1.7ml
In Elution pipes；
15) 30 μ l Elution Buffer, 200g centrifugation 2min are added, solution is made fully to be combined with column；
16) 14000g centrifuges 1min, by RNA uses without RNA deionized water dissolvings, for use.
3, the quality analysis (NanoDrop1000 spectrophotometers) of RNA sample
NanoDrop1000 spectrophotometers detect RNA sample, the sample requirement of RNA-seq sequencings：OD260/OD280 is
1.8-2.2。
By the RNA of said extracted into row agarose gel electrophoresis, Agilent Technologies
2100Bioanalyzer detects RNA sample quality, observes, takes pictures in gel imager, preserves image, it is considered that 28S:
It 18S >=2 can be preferable with preliminary judgement total serum IgE quality.
4, the extraction of miRNA and label
1) the miRNAs extraction agent boxes extracting of Ambion companies is used to obtain miRNA, concrete operations are according to corresponding instructions.
Sample is with T4RNA ligases markers step according to the method for Thomson.MiRNA labeling methods approximately as：1.4μg miRNA
With 500ng 5 '-phosphate-cytimidine-uracil cy3-3 ' (Dharmacon, Chicago, USA) and 2 unit T4RNA
Ligase (NEB, Ipswich, USA) is incubated 2 hours in 4 DEG C.Every part of miRNA sample is all provided with the corresponding negative control of equivalent.
2) mark RNA precipitated with the ethyl alcohol of 0.3M sodium acetates and 2.5 times of volumes, then with 15 μ l contain 3 × SSC,
The hybridization solution of 0.2%SDS and 15% formamide is resuspended, and all hybridization is repeated twice, hybridization LifterSlipTM (Erie,
PA USA) to ensure hybridization solution Uniform Flow between chip and cover plate.
3) hybridization chamber is placed on hybridization instrument BioMixerTMII (CapitalBio Corp, Beijing, China) in 42
DEG C water-bath is stayed overnight, and is washed twice with washing lotion.
5, miRNA chip operations：
MiRNA chips, using the miRNA chip of expression spectrum (single channel chip) of Boao Biological Co., Ltd, according to explanation
The instruction of book carries out the detection of miRNA express spectras.
6, result：
Analyze the testing result of miRNA chip expressions spectrum, it is known that miRNA-888-3p is in Degenerative disc disease patient
Tissue neutralizes the expression in the tissue of Healthy People, and there are significant differences, compared with the nucleus pulposus of Healthy People, in interverbebral disc regression
Property disease nucleus pulposus in miRNA-888-3p level significantly increase.
The miRNA-888-3p of embodiment 2QPCR verification differential expressions
1, miRNA-888-3p is selected to carry out large sample QPCR verifications according to the testing result of miRNA chips.According to implementation
Sample collection mode in example 1 selects Degenerative disc disease patient's nucleus pulposus sample and Healthy People nucleus pulposus sample each
60.
2, RNA extraction process is the same as embodiment 1.
3, reverse transcription：
1) 2 μM of miRNA reverse transcriptase primers of the total serum IgE template of 2 μ g and 1 μ l, 1 μ l, 2 μM of U6snRNA reverse transcriptions are drawn
Object, 1 μ l dNTP (10mM) mixtures and the deionized water mixing without RNase, final volume are 20 μ l, mixing.
2) 65 DEG C of incubation 5min.
3) it cools down on ice immediately.
4) following component is sequentially added：45 × buffer solutions of μ l, 2 μ l DTT (0.1M), the suppression of 1 μ l ribalgilases (RNase)
Preparation, by solution mixing.
5) 37 DEG C of incubation 5min.
6) 1 μ l M-MLV reverse transcriptases, mixing is added.
7) after 37 DEG C of reaction 1h, in 70 DEG C, 15min terminates reaction, -20 DEG C freeze it is spare.
In miRNA reverse transcriptase primers sequence such as sequence table shown in SEQ ID NO.2, U6snRNA reverse transcriptase primers sequence is such as
In sequence table shown in SEQ ID NO.3.
4, QPCR reacts：Using 25 μ l reaction systems, 3 parallel pipes are arranged in each sample, and all amplified reactions repeat
It is above to ensure the reliability of result three times.
1) following reaction system is prepared：
12.5 μ l of SYBR Green PCRs system, 1 μ l of forward primer (5 μM), 1 μ l of reverse primer (5 μM),
Template cDNA 2.0 μ l, 8.5 μ l of no enzyme water.Operations are carried out on ice.
2) amplification program is set：
95 DEG C of 10min, (95 DEG C of 20s, 60 DEG C of 55s) × 50 cycles.Using SYBR Green as fluorescent marker,
PCR reactions are carried out on Light Cycler fluorescence real-time quantitative PCR instrument.Expand the forward primer sequence of miRNA-888-3p such as
Shown in SEQ ID NO.4, reverse primer is as shown in SEQ ID NO.5.Using U6snRNA as reference gene, sense primer
Sequence is shown in SEQ ID NO.6；Downstream primer sequence is shown in SEQ ID NO.7.It is true by melt curve analysis analysis and electrophoresis
Determine purpose band, Δ Δ CT methods carry out relative quantification.
5, result
As shown in Figure 1, compared with the nucleus pulposus of Healthy People, miRNA- in Degenerative disc disease patient's nucleus pulposus
The expression of 888-3p significantly increases, consistent with miRNA chip results.
Expression of the embodiment 3miRNA-888-3p in Degenerative disc disease cell
1, cell culture
1) aseptically D-HANKS liquid is used to rinse 3 times the nucleus pulposus of acquisition.
2) nucleus pulposus is shredded into (about 1mm with scissors3Size), it is placed in sterile centrifugation tube.
3) it uses 0.25% trypsase to digest 20min in 37 DEG C, is shaken per 5min primary.
4) 800rpm centrifuges 5min, discards supernatant liquid.
5) 0.2% II Collagenase Type, 37 DEG C of digestion 4h, are filtered with 200 mesh screens.
6) filtrate centrifuges 5min with 800rpm, discards supernatant liquid.
7) DMEM/F12 culture mediums rinse, and 800rpm centrifuges 5min, is repeated 3 times.
8) culture bottle is inoculated in after cell count, the DMEM/F12 culture mediums containing 15% fetal calf serum and 1%P/S are as training
Nutrient solution, in 37 DEG C, 5%CO2Incubator in cultivate.
2、QPCR
2.1 cell total rnas extract：The extraction that cell total rna is carried out using the RNA extracts kits of QIAGEN companies, is pressed
Book instruction as directed carries out.
2.2QPCR：Step is the same as embodiment 2.
3, result
As shown in Fig. 2, compared with Healthy People nucleus pulposus cell, the nucleus pulposus cell miRNA-888-3p of Degenerative disc disease
Expression apparent increase (P<0.05).
Embodiment 4 studies influences of the miRNA-888-3p to Degenerative disc disease ability of cell proliferation
1, antisense oligonucleotides (anti-miRNA-888-3p) of the design synthesis for miRNA-888-3p
Anti- is synthesized by the design of Dalian treasured biotinylated biomolecule Technology Co., Ltd. according to the sequence information of miRNA-888-3p
MiRNA-888-3p and random controls sequence.
2, cell culture：The nucleus pulposus cell cultural method of Degenerative disc disease patient is the same as embodiment 3.
3, cell transfecting
Nucleus pulposus cell is divided into two groups, respectively inhibits negative control group (anti-NC), miRNA-888-3p inhibition groups
(anti-miRNA-888-3p).By negative control group and constituents for suppressing not Zhuan Ran anti-NC and anti-miRNA-888-3p, fortune
It is transfected with transfection reagent LipofectamineTM 2000, transfection method is with reference to specification.Anti-NC and anti-
The working concentration of miRNA-888-3p is 5 μM.48h collects each group cell and is used for subsequent experimental after transfection.
4, QPCR is tested
Cell total rna extracts and PCR step is the same as embodiment 3.
The results are shown in Figure 3, compared with inhibiting negative control group (anti-NC), miRNA-888-3p inhibition groups (anti-
MiRNA-888-3p the level of miRNA-888-3p) is remarkably decreased, and shows that anti-miRNA-888-3p can effectively inhibit
The expression of miRNA-888-3p.
5, cell proliferation experiment
The nucleus pulposus cell for transfecting 48h is digested to cell suspension using 0.25% pancreatin, with 5 × 104A/ml is inoculated in 96
Porocyte culture plates, per hole 0.1ml, after 60min, mtt assay surveys each hole 490nm wavelength absorbance values.With absorbance value size generation
The relative populations of table living cells.
6, result
MiRNA-888-3p inhibition group (anti-miRNA-888-3p) relative optical density number is 1.512 ± 0.213, is inhibited
Negative control group (anti-NC) relative optical density number is 0.215 ± 0.033.Compared with inhibiting negative control group (anti-NC),
MiRNA-888-3p inhibition group (anti-miRNA-888-3p) absorbance value significantly rises (P<0.05).Above-mentioned experimental result table
Bright miRNA-888-3p inhibits nucleus pulposus cell proliferation.
The explanation of above-described embodiment is only intended to understand the method and its core concept of the present invention.It should be pointed out that for this
For the those of ordinary skill in field, without departing from the principle of the present invention, several improvement can also be carried out to the present invention
And modification, these improvement and modification will also be fallen into the protection domain of the claims in the present invention.
Claims (7)
1.miRNA-888-3p inhibitor is preparing the application in treating Degenerative disc disease drug, which is characterized in that institute
The nucleotide sequence of miRNA-888-3p is stated as shown in SEQ ID NO.1.
2. application according to claim 1, which is characterized in that the miRNA-888-3p inhibitor can inhibit miRNA-
The expression of 888-3p or the function that miRNA-888-3p can be inhibited.
3. application according to claim 2, which is characterized in that the miRNA-888-3p inhibitor is miRNA-888-3p
Antisense oligonucleotides or antagonist.
4. according to claim 1-3 any one of them applications, which is characterized in that the drug further includes pharmaceutically acceptable
Carrier.
5. detecting the reagent of miRNA-888-3p expressions in preparing Degenerative disc disease early diagnosis kit
Using.
6. application according to claim 5, which is characterized in that the reagent includes the primer for miRNA-888-3p
And/or probe.
7. application according to claim 5, which is characterized in that the kit includes chip, and the chip includes solid phase
Carrier and the oligonucleotide probe being fixed on solid phase carrier, the oligonucleotide probe include specifically corresponding to
Some or all of miRNA-888-3p sequences.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102727909A (en) * | 2012-06-28 | 2012-10-17 | 中国人民解放军总医院 | Novel method for inhibiting primary liver cancer growth and metastasis |
| CN104704122A (en) * | 2012-04-20 | 2015-06-10 | 艾珀特玛治疗公司 | miRNA modulators of thermogenesis |
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| CN104704122A (en) * | 2012-04-20 | 2015-06-10 | 艾珀特玛治疗公司 | miRNA modulators of thermogenesis |
| CN102727909A (en) * | 2012-06-28 | 2012-10-17 | 中国人民解放军总医院 | Novel method for inhibiting primary liver cancer growth and metastasis |
Non-Patent Citations (1)
| Title |
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| miRNA-223,miRNA-181a在急性白血病中的表达及临床意义;刘佳等;《辽宁医学院学报》;20140430;第35卷(第2期);文章摘要 * |
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