CN109609652A - A kind of biomarker for diagnosing cancer of liver - Google Patents
A kind of biomarker for diagnosing cancer of liver Download PDFInfo
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Abstract
The invention discloses a kind of biomarkers relevant to hepatocellular carcinoma, the biomarker is AC239809.3, the present invention has found that AC239809.3 is related to hepatocellular carcinoma by experiment for the first time, it expresses up-regulation in patients with hepatocellular carcinoma, by the expression for detecting AC239809.3, it can be determined that whether subject suffers from hepatocellular carcinoma.
Description
Technical field
The invention belongs to biomedicine fields, are related to a kind of biomarker for diagnosing cancer of liver, are specifically related to give birth to
Object marker AC239809.3.
Background technique
Hepatocellular carcinoma (hepatocellular carcinoma, HCC) is one of most common malignant tumour, has and invades
The feature that attacking property is strong, the death rate is high ranks second (Esther, the Cidon.Systemic of cancer related mortality reason
treatment of hepatocellular carcinoma:Past,present and future[J].World
Journal of Hepatology,2017,9(18):797-807.).It is now recognized that hepatitis type B virus (HBV), hepatitis C
Viral (HCV), Alcoholic and evaluation of non-alcoholic cirrhotic patients, aflatoxin, obesity etc. are related to the generation of liver cancer.The hair of liver cancer
Interpretation of the cause, onset and process of an illness system is not yet completely clear, and research is thought with gene mutation, liver-cancer stem cell, microenvironment, non-coding RNA, metabolic disorder etc.
Related (Farazi P A, Depinho R A.Hepatocellular environment. [J] .Nature Reviews
Cancer,2006,carcinoma pathogenesis:from genes to 6(9):674-687.).Due to China's liver cancer
Low developed area is mostly occurred in, so that many patients are diagnosed Shi Duoyi and are in progressive stage or advanced stage, misses optimal operation
Opportunity.Liver transfer operation is considered as treatment liver cancer and potential liver disease most efficient method, but due to its surgery cost valuableness, and
There is a stringent indication, liver source relative shortage, although Technology of Liver Transplantation in China is highly developed in recent years, the overall mortality rate of liver cancer
It is not greatly improved yet.Targeted orally-administered chemotherapeutics Sorafenib is considered as the unique selection of advanced liver cancer patient, but its
Expensive, not all patient can bear, and late result is limited, so the prognosis of mid and late liver cancer patient is still very
Difference.In past decades, with science and technology progress, immunization therapy, gene therapy of tumour etc. achieve it is huge into
Therefore exhibition furthers investigate the gene profile of liver cancer cells and to find new therapy target most important.
The morbidity of liver cancer is one and is related to the complex process of a variety of regulations and molecular pathway, still not fully aware of at present.With
It is concentrated mainly on protein coding gene toward the Study on Molecular Mechanism to onset of liver cancer, because protein is considered all the time
It is the center of various biological processes.In contrast, RNA is considered as the intermediate link (mRNA) or platform of protein synthesis
(tRNA and rRNA).However as the development of high throughput sequencing technologies in recent years, it is considered as turning that people, which are gradually recognized over,
The non-coding RNA of record group " noise ", it is no matter quantitatively or functionally all abundanter and important than protein coding gene.It is long
Chain non-coding RNA (long non-coding RNA, lncRNA) is the RNA molecule that a kind of transcript length is more than 200nt, by
In lacking effective open reading frame, without the function of encoding albumen.Scientists have had realized that non-coding RNA,
Especially lncRNA is for cracking the significances of various sciences problems.Having more and more research reports at present, it has complexity
Biological function, lncRNA can epigenetics, transcription and transcription after etc. multiple level modulation gene expressions, also with disease
The development of disease is closely bound up.LncRNA differential expression in kinds cancer, multiple researchs confirm its generation that may participate in tumour, hair
Each stage of exhibition, invasion and transfer is the key factor that tumour occurs and is in progress.
Currently, molecular marker analyte detection has become standard in the diagnosis of breast cancer and colorectal cancer, it is therapeutic process
Important step.However, there is presently no unified hepatocellular carcinoma molecular marker standards.It is related to find hepatocellular carcinoma occurrence and development
LncRNA have great importance to the diagnosing and treating of tumour.
Summary of the invention
In order to make up for the deficiencies of the prior art, the purpose of the present invention is to provide a kind of biological markers for diagnosing cancer of liver
Object, by the expression for detecting biomarker, it can be determined that whether subject suffers from liver cancer, while the biomarker
It can also be used as the personalized treatment that molecular target is applied to liver cancer.
To achieve the goals above, the present invention adopts the following technical scheme:
The present invention provides application of the reagent of detection AC239809.3 in the product for preparing diagnosing hepatocellular carcinoma.
Further, the product includes being detected in sample by sequencing technologies, nucleic acid hybridization technique, nucleic acid amplification technologies
The reagent of the expression of AC239809.3.
Further, the reagent is selected from:
The probe of specific recognition AC239809.3；
Or the primer of specific amplification AC239809.3.
Further, the primer sequence of the specific amplification AC239809.3 is as shown in NO.1~2 SEQ ID.
The present invention provides a kind of product of vitro detection AC239809.3 expression, the product includes chip, examination
Agent box, nucleic acid film item.
Further, the chip includes the oligonucleotide probe of specific recognition AC239809.3；The kit includes
The primer of specific amplification AC239809.3 or the oligonucleotide probe of specific recognition AC239809.3；The nucleic acid film item
Oligonucleotide probe including specific recognition AC239809.3.
Further, the primer sequence of the specific amplification AC239809.3 is as shown in NO.1~2 SEQ ID.
The present invention provides the products of vitro detection AC239809.3 expression in the tool for preparing diagnosing hepatocellular carcinoma
In application.
The present invention provides application of the AC239809.3 in the computation model of building prediction hepatocellular carcinoma.
The present invention provides application of the AC239809.3 in the pharmaceutical composition of preparation treatment hepatocellular carcinoma.
Further, described pharmaceutical composition includes the inhibitor of AC239809.3, and the inhibitor can reduce
The expression of AC239809.3.
Detailed description of the invention
Fig. 1 is the expression figure using QPCR detection AC239809.3 gene in Tissues of Hepatocellular Carcinoma.
Specific embodiment
The method that the present invention is combined by using high-flux sequence and bioinformatics, screening in Tissues of Hepatocellular Carcinoma and
The lncRNA that differential expression is presented in corresponding cancer beside organism, then further verifies the differential expression by large sample
The relationship of lncRNA and hepatocellular carcinoma, and then determine a possibility that lncRNA is applied to diagnosis of hepatoma and treatment.This hair
It is bright to have found that AC239809.3 is raised in Expression In Hepatocellular Carcinoma for the first time by experiment.
AC239809.3 gene is located on No. 1 chromosome of people, and ensembl ID is in the ENSG00000227733 present invention
AC239809.3 includes wild type, saltant type or its segment.The AC239809.3 having disclosed at present there are 18 transcripts,
AC239809.3-201 (transcription ID:ENST00000599640.5), AC239809.3-214 (transcription ID:
ENST00000496508.1), AC239809.3-209 (transcription ID:ENST00000629702.3), AC239809.3-211 (turn
Record ID:ENST00000464343.6), AC239809.3-213 (transcription ID:ENST00000634551.1), AC239809.3-
215 (transcriptions ID:ENST00000611617.5), AC239809.3-203 (transcription ID:ENST00000611286.4),
AC239809.3-218 (transcription ID:ENST00000616018.1), AC239809.3-212 (transcription ID:
ENST00000628063.2), AC239809.3-217 (transcription ID:ENST00000620213.4), AC239809.3-207 (turn
Record ID:ENST00000599387.5), AC239809.3-206 (transcription ID:ENST00000596091.5), AC239809.3-
202 (transcriptions ID:ENST00000628902.1), AC239809.3-216 (transcription ID:ENST00000622642.4),
AC239809.3-210 (transcription ID:ENST00000596220.1), AC239809.3-208 (transcription ID:
ENST00000611452.4), AC239809.3-204 (transcription ID:ENST00000614606.4), AC239809.3-205 (turn
Record ID:ENST00000482381.2).Wherein, a kind of representative AC239809.3 gene such as AC239809.3-201 (transcription
ID:ENST00000599640.5 shown in sequence).
One skilled in the art will appreciate that when carrying out bioinformatic analysis to primitive sequencer result, it will usually will be sequenced
As a result it is compared with known gene, as long as sequencing fragment can compare on related gene, so that it may regard the gene as
Expression, therefore, in the gene for referring to differential expression, the different transcripts of the gene include simultaneously in the present invention.
It will be appreciated by those skilled in the art that the means of measurement gene expression are not importances of the invention.It can be
The expression of biomarker is detected on transcriptional level.The present invention can use any method known in the art measurement base
Because of expression.
Detection technique
LncRNA of the invention is detected using multiple nucleic acids technology known to persons of ordinary skill in the art, these skills
Art includes but is not limited to: nucleic acid sequencing, nucleic acid hybridization and nucleic acid amplification technologies.
The exemplary, non-limitative example of Nucleic acid sequencing techniques includes but is not limited to chain terminator (Sanger) sequencing and dye
Expect terminator sequencing.Those skilled in the art it will be recognized that due to RNA in cell less stable and in an experiment
It is more vulnerable to nuclease attack, therefore usually by RNA reverse transcription at DNA before sequencing.
The another exemplary non-limiting example of Nucleic acid sequencing techniques includes that (deep sequencing/high pass measures for next-generation sequencing
Sequence), high throughput sequencing technologies are a kind of sequencing technologies in synthesis based on unimolecule cluster, based on proprietary reversible termination chemistry
Reaction principle.The random fragment of the DNA of genome is attached to optically transparent glass surface when sequencing, these DNA fragmentations warp
After crossing extension and bridge amplification, hundreds of millions of clusters is formed in glass surface, each cluster is the list with thousands of parts of same templates
Then molecular cluster utilizes four kinds of special deoxyribonucleotides with fluorophor, skill is sequenced in synthesis by reversible
Template DNA to be measured is sequenced in art.
The exemplary, non-limitative example of nucleic acid hybridization technique include but is not limited in situ hybridization (ISH), microarray and
Southern or Northern trace.In situ hybridization (ISH) be it is a kind of use label complementary DNA or RNA chain as probe with
Position tissue a part or slice (original position) are the spy in entire tissue (full organization embedding ISH) if tissue is sufficiently small
The hybridization of anisotropic DNA or RNA sequence.DNA ISH can be used for determining the structure of chromosome.RNA ISH is for measuring and positioning group
Knit the mRNA and other transcripts (for example, ncRNA) in slice or full organization embedding.Usually to sample cell and tissue at
Reason increases the entrance of probe with fixation in situ target transcript.Probe hybridizes with target sequence at high temperature, then by extra spy
Needle is washed off.Use autoradiograph, fluorescence microscopy or immunohistochemistry respectively, in tissue with radiation, fluorescence or antigen
The probe of the kilobase marker of label is positioned and is quantified.Two or more can also be used by radioactivity by ISH or other are non-
The probe of radioactive label substance markers, to detect two or more transcripts simultaneously.
Southern and Northern trace is respectively used to detection specific DNA or RNA sequence.Make to extract from sample
DNA or RNA fracture, it is separated by electrophoresis on matrix gel, be then transferred on molecular filter.Make filter combine DNA or
RNA with and the complementary label probe of sequence of interest hybridize.Detection is integrated to the hybridization probe of filter.A kind of change of the program
Change form is reverse northern trace, wherein the substrate nucleic acid for being fixed to film is the set of isolated DNA fragmentation, and probe is
From tissue extraction and the RNA that is marked.
The exemplary, non-limitative example of nucleic acid amplification technologies includes but is not limited to: polymerase chain reaction (PCR) reverses
Record polymerase chain reaction (RT-PCR), the amplification (TMA) of transcriptive intermediate, ligase chain reaction (LCR), strand displacement amplification
(SDA) and the amplification based on nucleic acid sequence (NASBA).Those skilled in the art are it will be recognized that certain amplification technique (examples
Such as, PCR) it needs RNA reverse transcription before amplification at DNA (for example, RT-PCR), and other amplification techniques then direct cloning RNA
(for example, TMA and NASBA).
The polymerase chain reaction of commonly referred to as PCR uses denaturation, the annealing and primer extend of primer pair and opposite strand
Multiple circulations, exponentially increase target nucleic acid sequence copy number；The amplification of the transcriptive intermediate of TMA is (substantial constant
Temperature, multiple copies of target nucleic acid sequence are autocatalytically synthesized under conditions of ionic strength and pH, wherein target sequence is more
A RNA copy autocatalytically generates other copy；The ligase chain reaction of LCR uses miscellaneous with the adjacent area of target nucleic acid
The two groups of complementary DNA oligonucleotides handed over；Other amplification methods include for example: the commonly referred to as expansion based on nucleic acid sequence of NASBA
Increase；Use the amplification of rna replicon enzyme (commonly referred to as Q β replicase) amplification probe molecule itself；Amplification method based on transcription；
And the sequence amplification of self―sustaining.
The nucleic acid of non-amplification or amplification can be detected by any conventional means in the present invention.
Chip, nucleic acid film item, kit
The present invention provides the product of the expression of AC239809.3 gene in detection, the product includes (but unlimited
In) chip, nucleic acid film item or kit.Wherein chip includes: solid phase carrier；And it is orderly fixed on the solid phase carrier
Oligonucleotide probe, the oligonucleotide probe some or all of specifically correspond to shown in AC239809.3 sequence.
The solid phase carrier includes inorganic carrier and organic carrier, the inorganic carrier include but is not limited to have silicon carrier,
Glass carrier, ceramic monolith etc.；The organic carrier includes polypropylene film, nylon membrane etc..
The present invention provides the few cores for AC239809.3 that nucleic acid film item includes substrate and is fixed in the substrate
Thuja acid probe；The substrate can be any substrate suitable for immobilized oligonucleotide probe, such as nylon membrane, nitrocellulose
Film, polypropylene screen, sheet glass, silica gel chip, miniature magnetic bead etc.
The present invention provides a kind of kit, the kit includes the reagent for detecting AC239809.3 expression, Yi Jixuan
From one or more substances of the following group: container, operation instructions, positive control, negative control object, buffer, auxiliary agent or molten
Agent.
In kit of the invention can also have kit operation instructions, be described how using kit into
Row detection, and how tumor development to be judged using testing result, therapeutic scheme is selected.The component of kit can
To be packed in the form of aqueous medium or in the form of freeze-drying.In kit container appropriate typically at least include a kind of bottle,
Test tube, flask, PET bottle, syringe or other containers can carry out suitably wherein a kind of component can be placed, and preferably
Equal part.There are more than one group timesharing in kit, in kit generally also will comprising second, third or it is other additional
Container, wherein being positioned separately additional component.However, the component of various combination can be comprised in a bottle.The present invention
Kit generally also will include a kind of container for being used to accommodate reactant, sealing be used for commercial distribution.This container can wrap
The plastic containers of injection molding or blowing mould are included, wherein required bottle can be retained.
As a preferred embodiment, the kit is fluorescent quantificationally PCR detecting kit, the primer is applicable in
In SYBR Green, the detection of TaqMan probe, molecular beacon, double cross probe, combined probe.
In a further preferred embodiment, the PCR reaction solution in the kit is fluorescence quantitative PCR reaction solution,
And one step include fluorescent dye.
The present invention provides application of the AC239809.3 in the computation model of preparation prediction hepatocellular carcinoma.As skilled skill
Art personnel can be appreciated that, the measurement of two or more markers can be used to improve the diagnosis problem in investigation.It is biochemical
Marker can measure individually, or in one embodiment of the invention, they can be measured simultaneously, such as use chip
Or the array technique based on pearl.Then the independent concentration for interpreting biomarker, such as using individual retentions of every kind of marker,
Or they combine and are interpreted.
In the present invention, it can be implemented in various ways marker levels and certain possibility or risk association and realize
The step of getting up.Preferably, the mathematically measurement concentration of combination gene and one or more other markers, and by combined value
It associates with basic diagnosis problem.It can be by any suitable prior art mathematical method by the measurement group of marker levels
It closes.
Preferably, the mathematical algorithm applied in marker combination is a kind of logarithmic function.Preferably, using such mathematics
Algorithm or such logarithmic function the result is that single value.According to basic diagnosis problem, can easily by such value with it is for example a
Body associates about the risk of hepatocellular carcinoma or with the other intentional diagnostic uses for helping to assess hepatocellular carcinoma pine patient.With
A kind of preferred mode, such logarithmic function obtain as follows: individual segregation a) being entered group, such as normal person, has liver cell
The individual of cancer risk, the patient with hepatocellular carcinoma etc., b) identify that difference is aobvious between these groups by univariate analysis
The marker of work, c) logarithmic regressions analysis is to assess the independent difference values that can be used for assessing these difference groups of marker, and d)
Building logarithmic function carrys out composition independency difference value.In such analysis, marker is no longer independent, but represents one
A marker combination.
The present invention provides application of the AC239809.3 in the pharmaceutical composition of preparation treatment hepatocellular carcinoma, the drugs
Composition includes the inhibitor and pharmaceutically acceptable carrier of AC239809.3, and the inhibitor is selected from AC239809.3
Or its transcript is target sequence and the disturbing molecule for being able to suppress AC239809.3 gene expression or genetic transcription, comprising:
ShRNA (children purpura nephritis), siRNA (siRNA), dsRNA, Microrna, antisense nucleic acid, or can express or be formed and is described
ShRNA, siRNA, dsRNA, Microrna, antisense nucleic acid construction.Institute's pharmaceutically acceptable carrier includes (but simultaneously
It is not limited to) diluent, adhesive, surfactant, Humectant, absorption carrier, lubricant, filler, disintegrating agent.
The present invention is described in further detail with reference to the accompanying drawings and examples.Following embodiment is merely to illustrate this
It invents rather than limits the scope of the invention.Test method without specific conditions in embodiment, usually according to conventional strip
Part, or according to the normal condition proposed by manufacturer.
Embodiment 1 screens gene marker relevant to hepatocellular carcinoma
1, sample collection
The cancerous tissue and corresponding cancer beside organism's sample for collecting 35 hepatocellular carcinoma patients, therefrom randomly select
The acquirement of 5 progress high-flux sequences, the equal informed consent of patient, above-mentioned all samples passes through the same of the committee, organizational ethics
Meaning.
2, the preparation and quality analysis of RNA sample
It is shredded with scissors tissue, 1ml Trizol is added, shakes 1min on oscillator；Room temperature 10min, makes nucleoprotein
Body decomposes completely；Then 200 μ l chloroforms (chloroform) are added, cover tightly pipe lid, acutely shake 15s, after room temperature stands 10min, 4
DEG C, 11000rpm is centrifuged 15min；Water sample layer is transferred in a new centrifuge tube, 500 μ l isopropanols are added；It is mixed by inversion
Afterwards, room temperature stands after 10min 4 DEG C, and 11000rpm is centrifuged 15min；Liquid is carefully siphoned away with rifle, stays and is deposited in tube bottom, 1ml is added
75% ethyl alcohol shakes 5s on the oscillator, and washing precipitating is primary, and 4 DEG C, 8000rpm is centrifuged 5min；Then supernatant is carefully gone
Fall, drying precipitated 10min, suitable water dissolution precipitating 10min is added.
3, total serum IgE is quantitative and purity analysis
The RNA of extraction is subjected to agarose gel electrophoresis, using Nanodrop2000 to the concentration of mentioned RNA and purity into
Row detection, agarose gel electrophoresis detect RNA integrality, and Agilent2100 measures RIN value.5 μ of single requirement for construction data base RNA total amount
G, concentration >=200ng/ μ L, OD260/280 is between 1.8~2.2.
4, construction cDNA library
The rRNA in total serum IgE is removed using the Ribo-Zero kit of Epicentre；It will be complete using metal ion
Whole RNA is broken into the small fragment of 200bp or so at random；Utilize the TruseqTM RNA sample Prep Kit of Illumina
Carry out the building of cDNA library.
5, it is sequenced
CDNA library is sequenced using Illumina X-Ten microarray dataset.
6, high-throughput transcript profile sequencing data analysis
The lncRNA for being not easy to detect is deleted, carries out raw letter analysis using the DESeq2 in tool R-3.3.3.With
Cutadapt falls the base of quality < 20 to the 5 ' of reads and 3 ' Duan Jinhang trim, trim, and deletes N and be greater than 10%
reads；Then sequencing result is compared onto reference genome using tophat.Reference genome version used is hg38；
Cuffquant quantifies the expression quantity and normalization output of lncRNA, and cuffdiff compares control group with the table of disease group lncRNA
Up to difference, the screening criteria of differential expression lncRNA: FDR < 0.05, abs (log2FC)>1。
7, result
High-flux sequence is the results show that expression quantity of the AC239809.3 gene in Tissues of Hepatocellular Carcinoma is significantly higher than by cancer
It organizes (P < 0.001), AC239809.3 is prompted to can be used as the early diagnosis that possible detection target is applied to hepatocellular carcinoma.
The differential expression of embodiment 2QPCR sequence verification AC239809.3 gene
1, the 35 patient's liver cancer tissue samples and cancer beside organism's sample collected using front are poor to AC239809.3 gene
Different expression carries out large sample QPCR verifying.
2, RNA extraction step is referring to embodiment 1
3, QPCR is detected
1) reverse transcription reaction
A. by the total serum IgE template of 1 μ g and 2 10 × buffers of μ l, 2 μ l dATP (10mM), 0.5 μ l polyA polymerase,
0.5 μ l ribalgilase (RNase) inhibitor and deoxyribonuclease water (RNase free water) mixing, volume are finally
20 μ l, 37 DEG C of incubation 1h；
B. it is added 1 μ l 0.5 μ g/ μ l Oligo (dT) specific RT primer into reaction tube, 70 DEG C of incubation 5min, immediately
It sets and is incubated at least 2min on ice；
C. by reaction mixture and 45 × buffers of μ l, 1 μ l dNTP (10mM), 0.5 μ l M-MLV reverse transcriptase, 0.5 μ
L ribalgilase (RNase) inhibitor, 10 μ l polyA reaction mixtures and 4 μ l deoxyribonuclease water (RNase free
Water it) mixes, 42 DEG C of incubation 1h.
2) design of primers
QPCR amplimer, specific primer sequence are designed according to the coded sequence of AC239809.3 gene and GAPDH gene
It is as follows:
AC239809.3 gene:
Forward primer is 5 '-TCTTCTATCCTGGTGTTC-3 ' (SEQ ID NO.1)；
Reverse primer is 5 '-AGATGATTCAGATGATGTTAC-3 ' (SEQ ID NO.2).
GAPDH gene:
Forward primer is 5 '-AATCCCATCACCATCTTCCAG-3 ' (SEQ ID NO.3)；
Reverse primer is 5 '-GAGCCCCAGCCTTCTCCAT-3 ' (SEQ ID NO.4).
3) QPCR amplification is examined
25 μ l reaction systems: 12.5 μ l of SYBR Green polymerase chain reaction system are prepared, forward and reverse primer (5 μM) is each
1 μ l, template cDNA 2.0 μ l, ddH2O 8.5μl.Operations are carried out on ice.
Reaction condition: 95 DEG C of 10min, (95 DEG C of 15s, 60 DEG C of 60s) × 45 circulations.Using SYBR Green as fluorescence mark
Remember object, on Light Cycler fluorescence quantitative PCR instrument carry out PCR reaction, using GAPDH as reference gene, 60-95 DEG C into
The analysis of row solubility curve determines that purpose band, Δ Δ CT method carry out relative quantification by melt curve analysis analysis and electrophoresis.
4, result
QPCR result is as shown in Figure 1, compared with cancer beside organism, and AC239809.3 is raised in Expression In Hepatocellular Carcinoma, difference
With statistical significance (P < 0.05)；AC239809.3 expression up-regulation in all samples of detection, prompts AC239809.3
The application value with higher in the diagnosis of liver cancer.
The explanation of above-described embodiment is only intended to understand method and its core concept of the 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 scope of the claims in the present invention.
Sequence table
<110>No.2 Hospital, Hebei Medical Univ.
<120>a kind of biomarker for diagnosing cancer of liver
<160> 4
<170> SIPOSequenceListing 1.0
<210> 1
<211> 18
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 1
tcttctatcc tggtgttc 18
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<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 2
agatgattca gatgatgtta c 21
<210> 3
<211> 21
<212> DNA
<213>artificial sequence (Artificial Sequence)
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aatcccatca ccatcttcca g 21
<210> 4
<211> 19
<212> DNA
<213>artificial sequence (Artificial Sequence)
<400> 4
gagccccagc cttctccat 19
Claims (10)
1. detecting application of the reagent of AC239809.3 in the product for preparing diagnosing hepatocellular carcinoma.
2. application according to claim 1, which is characterized in that the product includes hybridizing skill by sequencing technologies, nucleic acid
The reagent of the expression of AC239809.3 gene in art, nucleic acid amplification technologies detection sample.
3. application according to claim 1, which is characterized in that the reagent is selected from:
The probe of specific recognition AC239809.3；Or
The primer of specific amplification AC239809.3.
4. application according to claim 3, which is characterized in that the primer sequence of the specific amplification AC239809.3 is such as
Shown in NO.1~2 SEQ ID.
5. a kind of product of vitro detection AC239809.3 expression, which is characterized in that the product includes chip, reagent
Box, nucleic acid film item.
6. product according to claim 5, which is characterized in that the chip includes the widow of specific recognition AC239809.3
Nucleotide probe；The kit includes the primer or specific recognition AC239809.3 of specific amplification AC239809.3
Oligonucleotide probe；The nucleic acid film item includes the oligonucleotide probe of specific recognition AC239809.3.
7. product according to claim 6, which is characterized in that the primer sequence of the specific amplification AC239809.3 is such as
Shown in NO.1~2 SEQ ID.
8. application of the described in any item products of claim 5-7 in the tool for preparing diagnosing hepatocellular carcinoma.
Application of the 9.AC239809.3 in the computation model of building prediction hepatocellular carcinoma.
10.AC239809.3 the application in the pharmaceutical composition of preparation treatment hepatocellular carcinoma.
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| CN103160580A (en) * | 2013-02-26 | 2013-06-19 | 中南大学 | Application method of long-chain non-coding ribonucleic acid (RNA) gene |
| CN106995858A (en) * | 2017-06-01 | 2017-08-01 | 北京泱深生物信息技术有限公司 | A kind of lncRNA related to liver cancer diagnosis and treatment |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103160580A (en) * | 2013-02-26 | 2013-06-19 | 中南大学 | Application method of long-chain non-coding ribonucleic acid (RNA) gene |
| CN106995858A (en) * | 2017-06-01 | 2017-08-01 | 北京泱深生物信息技术有限公司 | A kind of lncRNA related to liver cancer diagnosis and treatment |
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Application publication date: 20190412 |