CN106834528B

CN106834528B - A kind of biomarker for liver cancer diagnosis and treatment

Info

Publication number: CN106834528B
Application number: CN201710216076.1A
Authority: CN
Inventors: 王冬国; 陈佳玉
Original assignee: Taizhou Municipal Hospital
Current assignee: Taizhou Municipal Hospital
Priority date: 2017-04-01
Filing date: 2017-04-01
Publication date: 2019-10-29
Anticipated expiration: 2037-04-01
Also published as: CN106834528A

Abstract

The invention discloses a kind of biomarker for liver cancer diagnosis and treatment, the specific marker is LINC00511.Present invention firstly discovers that the differential expression of LINC00511 is related to the occurrence and development of liver cancer, in liver cancer patient, the expression of LINC00511 is raised, and prompts to intervene the new way that LINC00511 gene expression is likely to become liver cancer treatment.In order to further verify, the proliferation of liver cancer cells can be significantly reduced by the expression discovery that inhibitor reduces LINC00511 and cell forms clone's colony number.

Description

A kind of biomarker for liver cancer diagnosis and treatment

Technical field

The invention belongs to biomedicine fields, are related to a kind of biomarker for liver cancer diagnosis and treatment, are specifically related to give birth to Object marker LINC00511.

Background technique

Hepatocellular carcinoma (hepatocellular carcinoma, HCC) is the most common disease in adult human liver malignant tumour Disease, although the treatment technology of liver cancer is constantly promoted, such as perform the operation excision, liver transfer operation, chemicotherapy are survived for liver cancer patient overall 5 years Rate is not obviously improved yet so far.Liver cancer case fatality rate is high, poor prognosis one of the major reasons are exactly the molecule machine of liver cancer genesis and development System is still known little about it at present.However, having been demonstrated that how adjoint the formation of liver cancer is with the progress of INVENTIONModern cell molecular biology A series of exception of molecules and signal path.Therefore, the mechanism of liver cancer genesis and development is explored from molecular level, and characteristic is divided Son is used as anti-tumor target, can open up new approach for clinical prevention and treatment liver cancer.

With the rapid development of microarray technology and Nucleic acid sequencing techniques, scientist is had found only 2% in human genome Sequential coding generate protein, protein coding gene number deficiency 30,000, and remaining nearly 98% genome sequence turns Record produces a large amount of and miscellaneous non-coding RNA, these ncRNA are the important sets of complicated regulated and control network in body At part.

Long-chain non-coding RNA (long noncoding RNA, lncRNA) is that one kind can be sent out during various biological Wave the macromolecular ncRNA of regulating and controlling effect.Its is widely distributed, and length is typically greater than 200 bases, due to lacking effective open reading The ability of frame (ORF) and nothing or few coding albumen.As a uncharted field in molecular biology, lncRNA be with The form of RNA played on many levels controlling gene expression effect, mainly epigenetic regulation, transcriptional control and Three levels of post-transcriptional control are regulated and controled.As the important component of mammalian transcription group, the function of lncRNA is current It need to be furtherd investigate.Initial lncRNA is considered as the by-product of rna plymerase ii transcription, is that subgenomic transcription " is made an uproar Sound ", " dark matter " do not have biological function.But result of study in recent years is shown, lncRNA is in cell normal physiological activity Important function and participate in the occurrence and development of kinds of tumors and other diseases.LncRNA not only participates in transport in core, turns Record in the cellular physiological events such as regulation, protein degradation, genomic imprinting and X chromosome silencing, also with breast cancer, prostate cancer, The pathogenesis of the diseases such as non-small cell lung cancer, lymphatic leukemia is related.LncRNA can pass through Genomic Imprinting, chromatin weight Structure shears the mechanism such as regulation, mRNA degradation and translational control to implement its function.Although lncRNA correlative study takes in recent years Progress was obtained, but its function and mechanism of action still need further to be explored.Research lncRNA main method and tool include chip, RNA sequencing, real-time quantitative PCR, Northern blotting, in situ hybridization, RNAi, RIP and Bioinformatics Prediction etc..It grinds The development and utilization for studying carefully technology is very crucial for the research of biological mechanism.

LncRNA is widely present in Various Tissues, and expression has tissue specificity and Space-time speciality.Organizing specific Property refer to that the lncRNA expression quantity between different tissues is different；Space-time speciality refers to the different hairs in identical tissue or organ It educates the stage, lncRNA expression quantity therein can also change.Someone to the expression specificity of 1,300 lncRNA of mouse brain into Row research finds there is different expression patterns in the different parts of tissue development process mesencephalic tissue, further proves The Space-time speciality of lncRNA expression.Meanwhile lncRNA also has the expression pattern of specificity in tumour and other diseases.Mesh It is preceding to be still in the starting stage for research of the lncRNA in cancer field, effect of the lncRNA in liver cancer is sought with important Meaning.

Summary of the invention

In order to make up for the deficiencies of the prior art, an object of the present invention provides a kind of product for diagnosing early liver cancer, makes Patient is just treated in early days, and then improves survival rate and quality of life.

The second object of the present invention provides a kind of molecular marker, the detection or treatment index as clinical application.

The third object of the present invention provides a kind for the treatment of means and pharmaceutical composition, realizes the accurate molecular therapy of liver cancer.

To achieve the goals above, the present invention adopts the following technical scheme:

The present invention provides a kind of purposes of LINC00511 gene, are used to prepare the product of diagnosing liver cancer.

Further, the LINC00511 expresses up-regulation in liver cancer patient.

Further, the product can by RT-PCR, real-time quantitative PCR, in situ hybridization, northern blotting, The expression of chip or high-flux sequence detection of platform LINC00511.

Wherein, the product with RT-PCR diagnosing liver cancer includes at least drawing for a pair of of specific amplified LINC00511 gene Object；The product with real-time quantitative PCR diagnosing liver cancer includes at least the primer of a pair of of specific amplified LINC00511 gene；Institute Stating with the product of in situ hybridization diagnosing liver cancer includes: probe with the nucleic acid array hybridizing of LINC00511 gene；It is described to use chip The product of diagnosing liver cancer includes the probe with the nucleic acid array hybridizing of LINC00511 gene.

Further, the product with real-time quantitative PCR diagnosing liver cancer includes at least a pair of of specific amplification LINC00511 The primer of gene, the primer is as shown in SEQ ID NO.4 and SEQ ID NO.5 in a specific embodiment of the invention.

The present invention provides a kind of product of diagnosing liver cancer, the product can pass through LINC00511 in detection sample Expression carrys out diagnosing liver cancer." sample " includes but is not limited to cell, tissue, internal organs, body fluid (blood, lymph Deng), digestive juice, expectoration, alveole bronchus cleaning solution, urine, excrement etc..Preferably, the sample is tissue, blood.

Further, the product includes chip, preparation or kit；Wherein, the chip includes solid phase carrier and consolidates It is scheduled on the oligonucleotide probe of solid phase carrier, the oligonucleotide probe includes the needle for detecting LINC00511 transcriptional level To the oligonucleotide probe of LINC00511；The kit includes the primer or core for detecting LINC00511 transcriptional level Piece.

The various common used materials in genetic chip field, such as, but not limited to nylon membrane can be used in solid phase carrier, through active group Slide or silicon wafer, unmodified slide, plastic sheet of group's (such as aldehyde radical, amino) modification etc..

" probe " refers to can be with the molecule in conjunction with the particular sequence of another molecule or subsequence or other parts.Unless otherwise finger Out, term " probe " is often referred to match by complementary base and combine with another polynucleotides (often referred to as " target polynucleotide ") Polynucleotide probes.According to the preciseness of hybridization conditions, probe energy and with the probe lack sufficient sequence complementarity target it is more Nucleotide combines.Probe can make direct or indirect label, and range includes primer.Crossing system includes, but are not limited to: molten Liquid phase, solid phase, mixed phase or in situ hybridization measuring method.

The probe has the base sequence with the specific base sequence complementary of target gene.Here, so-called " complementation ", As long as hybridization, can not be complete complementary.These polynucleotides have usually relative to the specific base sequence 80% or more, preferably 90% or more, more preferable 95% or more, particularly preferred 100% homology.These probes can be DNA, It is also possible to RNA, furthermore it is possible to pass through PNA (Polyamide nucleic in part of it or whole nucleotides Acid, peptide nucleic acid), LNA (registered trademark, locked nucleic acid, Bridged Nucleic Acid, Cross-linked core Acid), ENA (registered trademark, 2 '-O, 4 '-C-Ethylene-bridged nucleic acids), GNA (Glycerol Nucleic acid, glycerol nucleic acid), the artificial replacement nucleic acid such as TNA (Threose nucleic acid, threose nucleic acid) obtains Polynucleotides.

Gene detecting kit or genetic chip can be used for detecting multiple including LINC00511 gene in the present invention The expression of gene (for example, multiple genes relevant to liver cancer) detects multiple markers of liver cancer simultaneously, can be big The big accuracy rate for improving diagnosing cancer of liver.

The present invention provides a kind of purposes of LINC00511 gene, it is used to prepare prevention or treats the pharmaceutical composition of liver cancer Object.

Further, described pharmaceutical composition includes the inhibitor of LINC00511 functional expression, and the inhibitor can press down LINC00511 processed or be related to the upstream LINC00511 or downstream pathway substance expression.

Further, the inhibitor is the siRNA for LINC00511.

The present invention provides a kind of pharmaceutical composition, described pharmaceutical composition includes the suppression of LINC00511 functional expression Preparation, the inhibitor can work on DNA level or RNA (i.e. gene product) level.

Further, described pharmaceutical composition further includes its other medicine with the inhibitor compatibility of LINC00511 functional expression Class and pharmaceutically acceptable carrier and/or auxiliary material.

Further, the carrier includes (but being not limited to): diluent, excipient such as lactose, sodium chloride, glucose, urine Element, starch, water etc., filler such as starch, sucrose etc.；Adhesive such as simple syrup, glucose solution, starch solution, cellulose spread out Biology, alginates, gelatin and polyvinylpyrrolidone；Wetting agent such as glycerol；Disintegrating agent such as dried starch, sodium alginate, kelp are more Icing Sugar end, agar powder, calcium carbonate and sodium bicarbonate；Sorbefacient quaternary ammonium compound, lauryl sodium sulfate etc.；Surface is living Property agent such as polyoxyethylene sorbitan aliphatic ester, lauryl sodium sulfate, glyceryl monostearate, hexadecanol etc.；It causes Humectant such as glycerol, starch etc.；Absorption carrier such as starch, lactose, bentonite, silica gel, kaolin and soap clay etc.；Lubricant is such as sliding Mountain flour, calcium stearate and magnesium, polyethylene glycol, boric acid powder etc..

The present invention provides a kind of purposes of LINC00511 gene, for screening the potential substance of prevention or treatment liver cancer.

Further, screening prevention or the step for the treatment of liver cancer potential substance, include:

Candidate substances processing expression or the system containing LINC00511 gene；With

Detect the expression of LINC00511 gene in the system；

Wherein, if the candidate substances can reduce LINC00511 gene expression or activity (preferably significantly reduce, such as it is low 20% or more, preferably low 50% or more, more preferably low 80% or more) then shows that the candidate substances are prevention or treatment liver cancer Potential substance.

Further, system recited above includes but is not limited to: cell system, subcellular system, solution system, tissue System, organ systems or animal system.

In the present invention, siRNA may include that partially purified RNA, quite pure RNA, the RNA of synthesis or recombination produce Raw RNA and by addition, delete, replacement and/or to change one or more nucleotide different from naturally occurring RNA The RNA through changing.Such change may include addition non-nucleotide material to the end of such as siRNA (one or more It is a) or to nucleotide inside the one or more of siRNA, including making siRNA resist the modification of nuclease digestion.

The present invention is when screening effective siRNA sequence, by largely comparing analysis, to find out optimal effective piece Section.In a specific embodiment of the invention, the present inventor's design has synthesized a variety of siRNA sequences, and they are passed through respectively Transfection reagent transfecting hepatoma cells system is verified, and as a result detects the preferable disturbing molecule of interference effect, they are respectively provided with Sequence shown in SEQ ID NO.12, SEQ ID NO.13 is further tested in cellular level, is as a result proved for cell reality Inhibit efficiency very high for testing.

Nucleic acid inhibitor of the invention such as siRNA can be with chemical synthesis, can also be by a recombinant nucleic acid structure Expression cassette is prepared after being transcribed into single stranded RNA.The nucleic acid inhibitors such as siRNA, can be by using transfection reagent quilt appropriate It is transported into the cell, or multiple technologies known in the art also can be used and be transported into the cell.

In the present invention, pharmaceutical composition can be used different additives and be prepared, for example, buffer, stabilizer, Bacteriostatic agent, isotonic agent, chelating agent, pH controlling agent and surfactant.

Buffer may include boric acid, phosphoric acid, acetic acid, citric acid, glutamic acid and corresponding salt (their alkali metal or alkali Property rare earth metal salt, such as sodium salt, sylvite, calcium salt and magnesium salts).Isotonic agent includes potassium chloride, sodium chloride, sugar and glycerol.Chelating Agent includes sodium ethylene diamine tetracetate and citric acid.Bacteriostatic agent includes but is not limited to the benzylalcohol of effective concentration (such as < 1%w/v), benzene Phenol, metacresol, methaform, methyl p-hydroxybenzoate and/or propylparaben.Stabilizer includes human serum egg White, l-amino acid, sugar and cellulose derivative.L-amino acid can also include appointing in glycine, cysteine and glutamic acid Meaning one.Carbohydrate includes monosaccharide, such as glucose, mannose, galactolipin, fructose etc.；Sugar alcohol, such as mannitol, inositol, wood Sugar alcohol etc.；Disaccharides, such as sucrose, maltose, lactose etc.；Polysaccharide, for example, glucan, hydroxypropul starch, vulcanization chondroitin, thoroughly Bright matter acid etc. and their derivative.Cellulose derivative includes methylcellulose, ethyl cellulose, hydroxyethyl cellulose, hydroxyl Propyl cellulose, hypromellose and sodium cellulose glycolate.Surfactant includes ion or non-ionic surface active Agent, such as polyethylene glycol oxide Arrcostab, sorbitan monoacyl ester, fatty glyceride.

Drug of the invention may also include pharmaceutically acceptable coating material, fast decoupled coating Material, coloring agent, enteric polymer, plasticizer, water-soluble polymer, insoluble polymer, dyestuff, pigment, other collapse Powder.Common fast decoupled coating material includes OPADRY；Enteric polymer includes methylacrylic acid polymer, phosphorus hydroxypropyl first Base cellulose phthalate, hypromellose acetic acid esters, hypromellose succinate, hydroxyl first ethyl cellulose Element, cellulose acetophthalate；Plasticizer includes polyethylene glycol (PEG), propylene glycol etc..

Pharmaceutical composition Orally-administrable of the present invention, parenteral administration are given by sucking spray delivery, part Medicine, rectally, nasal administration, cheek administration, vagina administration are administered by the storage medicine device of implantation.It is preferred that oral administration or injection Administration.Pharmaceutical composition of the invention contains any commonly employed nontoxic pharmaceutical acceptable carrier, auxiliary material or excipient.In certain situations Under, medicinal acid, alkali or buffer can be used to adjust the pH of preparation to improve the stabilization of prepared compound or its form of administration Property.Terms used herein parenteral route include subcutaneous, intradermal, intravenous, intramuscular, intra-articular, intra-arterial, intrasynovial, in breastbone, In bringing up, in damage location and intracranial injection or infusion techniques.As long as destination organization can be reached, pharmaceutical composition of the present invention Receptor can be given by any approach.

Pharmaceutical composition of the present invention can be administered orally in the form of any peroral dosage form, including but not limited to capsule, piece Agent, emulsion and water slurry, dispersing agent and solution.For oral tablet, common carrier includes lactose and cornstarch.It is general to go back Lubricant such as magnesium stearate is added.In order to be administered with capsules per os, applicable diluent includes lactose and anhydrous corn Starch.When water slurry and/or lotion is administered orally, active component can be suspended or dissolved in oily phase, and and emulsifier And/or suspending agent merges.If necessary, some sweeteners and/or corrigent and/or colorant can be added.Where appropriate, can The dosage unit preparations packet micro-capsule that will be used to be administered orally.For example, by the way that particulate matter is coated or is wrapped in polymer, wax etc. It buries, the preparation can also be prepared and extended or maintained release.Pharmaceutical composition of the invention can be used for reducing endogenic LINC00511 is overexpressed, by reducing the expression of LINC00511, thus liver caused by treatment is raised because of LINC00511 expression Cancer.

In the present invention, the compound of LINC00511 expression can will be inhibited as naked RNA and delivery of agents together as core Sour (such as recombinant plasmid or viral vectors) is applied to subject, and the nucleic acid includes the sequence for inhibiting LINC00511 expression.Delivering Reagent can be lipophilic agent, polycation, liposome etc..

Drug of the invention can also with the drug combination of other treatment liver cancer, other therapeutic compound can with it is main Active constituent is administered simultaneously, or even is administered simultaneously in same composition.Can also with individual composition or with main work The property different dosage form of ingredient individually gives other therapeutic compounds.The Fractional of main component can be with other treatments Property compound is administered simultaneously, and other dosage can be administered alone.Over the course for the treatment of, can according to the severity of symptom, The frequency of recurrence and the physiologic response of therapeutic scheme adjust the dosage of pharmaceutical composition of the present invention.

Pharmaceutical composition of the present invention can also be administered in the form of Liposomal delivery systems, such as small monolayer vesicle, big list Layer vesica and multi-layer vesicles.Liposome can be formed there are many phosphatide, such as cholesterine, stearic amine or phosphatidyl choline.Liposome The blood halflife of gene product or nucleic acid can be increased.It can (it generally includes neutrality from the lipid of the formation vesicles of standard Or negatively charged phosphatide and sterol such as cholesterol) formed for suitable liposome of the invention.In general, by consider into The factors such as half-life period are directly chased after in the size and blood flow of purpose liposome, instruct the selection of lipid.

Liposome for this method may include by the cytotropic ligand molecular of liposome target.It is generally deposited in conjunction in cancer cell The ligand of receptor be for example preferred in conjunction with the monoclonal antibody of tumor-cell antigen.It can also be to for lipid of the invention Body is modified, to avoid by mononuclear phagocyte system and reticuloendothelial system removing.The liposome of such modification, which has, to be deposited The opsonic action for being on surface or being integrated into liposome structure inhibits part.Preferably, liposome may include opsonic action Inhibit both part and ligand.

Pharmaceutical composition of the present invention can be configured to ointment, cream, suspension with the pharmaceutical composition of local administration Agent, lotion, powder, solution, paste, gelling agent, spray, aerosol or finish.

It would be recognized by those skilled in the art that practicability of the invention is not limited to marker gene of the invention The gene expression of any specific variants is quantified.In the mankind, which has 17 annotation transcripts (or splice variant): LINC00511-018 (the transcript ID in Ensembl is ENST00000581549.1), the long 1716bp of long 2265bp The LINC00511-009 of LINC00511-002 (the transcript ID in Ensembl is ENST00000453722.6), long 1696bp The LINC00511-011 of (the transcript ID in Ensembl is ENST00000457958.6), long 908bp (turn in Ensembl Recording this ID is ENST00000578206.1), (the transcript ID in Ensembl is by the LINC00511-013 of long 757bp ENST00000583460.1), (the transcript ID in Ensembl is the LINC00511-016 of long 657bp ENST00000581183.1), (the transcript ID in Ensembl is the LINC00511-005 of long 561bp ENST00000580500.5), (the transcript ID in Ensembl is the LINC00511-007 of long 538bp ENST00000577828.5), (the transcript ID in Ensembl is the LINC00511-017 of long 536bp ENST00000580199.5), (the transcript ID in Ensembl is the LINC00511-012 of long 520bp ENST00000577773.1), (the transcript ID in Ensembl is the LINC00511-010 of long 475bp ENST00000580948.1), (the transcript ID in Ensembl is the LINC00511-003 of long 467bp ENST00000580861.1), (the transcript ID in Ensembl is the LINC00511-004 of long 436bp ENST00000580175.1), (the transcript ID in Ensembl is the LINC00511-006 of long 391bp ENST00000584749.1), (the transcript ID in Ensembl is the LINC00511-014 of long 379bp ENST00000581801.5), (the transcript ID in Ensembl is the LINC00511-015 of long 355bp ENST00000579631.1), (the transcript ID in Ensembl is the LINC00511-008 of long 141bp ENST00000582712.1).In specific embodiments, LINC00511 gene product refers to long transcript.As unrestricted Property example, the cDNA sequence that marker gene can have SEQ ID NO.1, SEQ ID NO.2 or SEQ ID NO.3 specified. In some embodiments, have with the same or similar cDNA sequence of listed sequence at least 70%, it is such as above-mentioned listed Sequence at least 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or at least 99% the same or similar cDNA sequence.

In the present invention, about " functional expression " of LINC00511, it is intended that the transcription of functional gene product and/or Translation.For the non-protein encoding gene as LINC00511, " functional expression " can lack of proper care at least two levels.The One, on DNA level, such as through the missing or destruction of gene, or there is no transcription (all to prevent in both cases The synthesis of relevant gene product).The missing of transcription can be for example by the variation of epigenetic (such as DNA methylation) or by function Delete mutation causes." afunction " or " LOF " mutation as used herein is enhanced relative to protein is assigned Or for new active function gain mutation, prevent, reduce or eliminate the mutation of the function of gene product.Functionality lacks Mistake can be caused by extensive mutation type, including but not limited to the deletion of whole gene or Gene Partial, splice site mutation, by Small insertion and frameshift mutation caused by deleting, nonsense mutation, instead of essential amino acid missense mutation and prevent product The mutation of correct cellular localization.If this definition also includes the mutation of the promoter or regulatory region of LINC00511 gene --- These mutation disturbances function of gene.Null mutation is the complete LOF mutation for destroying gene product function.One allele In null mutation will usually make expression reduce by 50%, but may to the function of gene product have seriously affect.It is worth It is noted that functional expression can also lack of proper care because of function gain mutation: by assigning protein new activity, albumen The normal function of matter is lacked of proper care, and the functional activity albumen of expression is reduced.Vice versa, and functional expression can be for example multiple by gene System is increased by lacking DNA methylation.Functional expression can also lack of proper care because of function gain mutation: pass through imparting The new activity of protein, the normal function imbalance of protein, the functional activity albumen of expression are reduced.Vice versa, functional table Up to can for example by gene duplication or by lack DNA methylation increase.

Second, on rna level, such as by lacking effective translation-for example because mRNA's is unstable (such as logical Cross UTR variant), it can cause to be degraded in the premessenger RNA of translation of transcript.Or effectively transcribed by lacking, such as because Mutation is induction of new splice variant.

Pharmaceutical composition of the invention can be by effectively amount be administered in pharmacy, term " pharmacy of the invention Upper effective amount " is referred to be applicable to reasonably the receiving benefits of therapeutic treatment or prevention/risk-benefit risks and be enough to treat or prevent The amount of disease, can be according to including the severity of disease, the activity of drug, the age of patient, weight, health, gender, patient couple The susceptibility of drug, the administration time of the used present composition, administration route and discharge ratio, treatment time and institute Well known element is determined in the element of the composition cooperation of the invention or the drug used simultaneously that use and other medical domains Determine effective dose level.Pharmaceutical composition of the invention can be used as individual therapeutic agent and be administered, or with other therapeutic agents And land used is administered, and can in turn or be simultaneously administered with previous therapeutic agent.In addition, can single or multiple times into Row administration.Importantly, needing to take in above-mentioned element, and maximum effect can be obtained with least amount without side-effects The amount of fruit is administered.

Term " treatment " refers to not for the purpose of curing, but slows down pathological condition or illness or prevent that (reduction) targets Recurrence.If after the therapeutic agent for receiving therapeutically effective amount, successfully by " treatment ", patient shows can observe patient And/or the mensurable sign of one or more specified diseases and the reduction of symptom or disappearance.For example, cancer cell count significantly subtracts Less or cancer cell disappears, and reduces tumor size；Inhibit and (that is, slowing down to a certain extent, and preferably stop) metastases；Certain Inhibit tumour growth in kind degree；Make to reduce and/or mitigate to a certain extent associated with particular cancers one or more The time of symptom increases；The morbidity and mortality of reduction, and quality of making the life better.The sign of disease or the mitigation energy of symptom It is enough patient perceivable.Treatment may be implemented to react-be defined as all signs disappearance of cancer or part reaction-tumour completely Size reduces, and the ratio preferably reduced is more than 50%, more preferable 75%.If patient perceptions, to stable disease, patient is also regarded To obtain medical treatment.

The advantages of the present invention:

Present invention firstly discovers that a kind of lncRNA relevant to liver cancer genesis and development, by the expression for detecting the lncRNA Level can detect early liver cancer, to treat to early liver cancer patient, improve the life quality of patient.

The present invention provides a kind of accurate medical procedures, by inhibiting the expression of LINC00511 in patient, to control Treat liver cancer patient.

The present invention provides a kind of therapeutic agent of liver cancer, which can treat liver caused by LINC00511 is overexpressed Cancer.

Detailed description of the invention

Fig. 1 shows the expression figure using QPCR detection LINC00511 in liver cancer patient；

Fig. 2 shows the expression figure using QPCR detection LINC00511 in liver cancer cells；

Fig. 3 is the expression influence diagram using QPCR detection transfection siRNA to the LINC00511 in liver cancer cells；

Fig. 4 is the influence diagram using CCK8 detection LINC00511 cell proliferation；

Fig. 5 shows LINC00511 to the influence diagram of the Clone formation colony of cell.

Specific embodiment

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, such as Sambrook et al., molecular cloning: laboratory manual (New York:Cold Spring HarborLaboratory Press, 1989) condition described in, or according to the normal condition proposed by manufacturer.

Embodiment 1 screens gene marker relevant to liver cancer

1, sample collection

Each cancerous tissue for collecting 10 liver cancer patients and cancer beside organism, the equal informed consent of patient, above-mentioned all samples Obtain the agreement for passing through the committee, organizational ethics.

2, the preparation of RNA sample

The extraction of tissue RNA is carried out using the tissue RNA extracts kit of QIAGEN, the specific steps of by specification carry out Operation.

3, reverse transcription and label

With Low RNA Input Linear Amplification Kit by mRNA reverse transcription at cDNA, while using Cy3 Labelling experiment group and control group respectively.

4, hybridize

Genetic chip uses Kang Cheng biology-Human lncRNA Array, carries out by the step of chip operation instructions miscellaneous It hands over.

5, data processing

Chip Agilent scanner scanning after hybridization, resolution ratio are 5 μm, and scanner is automatically with 100% and 10%PMT Each scanning 1 time, 2 times result Agilent software merges automatically.Scan image data is using at Feature Extraction Reason analysis, obtained initial data application Bioconductor program bag carry out follow-up data processing.Last Ratio value is experiment Group and control group.Differential gene screening criteria: ratio >=4 are up-regulation gene, and ratio≤0.25 is down-regulated gene.

6, result

Compared with cancer beside organism, expression of the LINC00511 in liver cancer tissue is significantly raised.

The differential expression of 2 QPCR sequence verification LINC00511 gene of embodiment

1, large sample QPCR verifying is carried out to LINC00511 gene differential expression.According to the sample collection side in embodiment 1 Formula collects liver cancer tissue and each 60, cancer beside organism's sample.

2, RNA extraction step is the same as embodiment 1.

3, reverse transcription:

Using 25 μ l reaction systems, each sample takes 1 μ g total serum IgE as template ribonucleic acid, is separately added into PCR pipe following Component: DEPC water, 5 × RT Buffer, 10mM dNTP, 0.1mM DTT, 30 μM of Oligo dT, 200U/ μ l M-MLV, Template ribonucleic acid.42 DEG C of incubation 1h, 72 DEG C of 10min, of short duration centrifugation.

(3) QPCR amplification is examined

Design of primers:

The primer sequence of LINC00511 gene are as follows:

Forward primer: 5 '-GCTCAAGTTCCTGACATAA-3 ' (SEQ ID NO.4)

Reverse primer: 5 '-GCGTTGTGTTAGGCATTA-3 ' (SEQ ID NO.5)

The primer sequence of house-keeping gene GAPDH are as follows:

Forward primer: 5 '-CCGGGAAACTGTGGCGTGATGG-3 ' (SEQ ID NO.6)

Reverse primer: 5 '-AGGTGGAGGAGTGGGTGTCGCTGTT-3 ' (SEQ ID NO.7)

Using 25 μ l reaction systems, 3 parallel pipes are arranged in each sample, and all amplified reactions are repeated three times above to protect Demonstrate,prove the reliability of result.

Prepare following reaction system: 12.5 μ l of SYBR Green polymerase chain reaction system, forward and reverse primer (5 μM) Each 1 μ l, template cDNA 2.0 μ l, no 8.5 μ l of enzyme water.Operations are carried out on ice.Amplification program are as follows: 95 DEG C of 60s, (95 DEG C 15s, 60 DEG C of 15s, 72 DEG C of 45s) × 35 circulations.It is glimmering in Light Cycler using SYBR Green as fluorescent marker PCR reaction is carried out on light real-time PCR, determines that purpose band, Δ Δ CT method carry out by melt curve analysis analysis and electrophoresis Relative quantification.

3, statistical method

Experiment is completed according to being repeated 3 times, and result data is indicated in a manner of mean+SD, Using SPSS18.0 statistical software come for statistical analysis, difference between the two is examined using t, it is believed that is had as P < 0.05 It is statistically significant.

4, result

As a result as shown in Figure 1, compared with cancer beside organism, LINC00511 gene is raised in Expression In Hepatocellular Carcinoma, difference tool Statistically significant (P < 0.05), it is consistent with RNA-sep result.

Differential expression of the 3 LINC00511 gene of embodiment in liver cancer cell lines

1, cell culture

HepG2 cell lines, Huh7 and normal liver cell system HL-7702, to contain 10% fetal calf serum and 1%P/S Culture medium DMEM in 37 DEG C, 5%CO₂, relative humidity be 90% incubator in cultivate.It changes within 2-3 days liquid 1 time, uses The 0.25% trypsase conventional digestion passage containing EDTA.

2, the extraction of RNA

1) culture is terminated when cell is merged up to 80-90%, 0.25% trypsin digestion is collected cell and managed in 1.5m1EP In, lm1Trizol is added in every pipe and slowly shakes smudge cells, places 10min on ice.

2) deproteinized removes DNA: 0.2ml chloroform is added in each 1.5m1EP pipe, rocks 15s, is placed at room temperature for 10min.4 DEG C, 12000rpm is centrifuged 15min.

Remaining operation step is the same as RNA extraction process in tissue.

3, reverse transcription

Specific steps are the same as embodiment 2.

4, statistical method

5, result

As a result as shown in Fig. 2, compared with normal liver cell system, LINC00511 gene is in hepatocellular carcinoma H22, Huh7 Expression is raised, and difference has statistical significance (P < 0.05), consistent with RNA-sep result.

The silencing of 4 LINC00511 gene of embodiment

1, cell culture

HepG2 cell lines, with the culture medium DMEM containing 10% fetal calf serum and 1%P/S in 37 DEG C, 5%CO₂、 It is cultivated in the incubator that relative humidity is 90%.It changes within 2-3 days liquid 1 time, trypsase conventional digestion of the use 0.25% containing EDTA Passage.

2, siRNA is designed

For the siRNA sequence of LINC00511 gene:

Negative control siRNA sequence (siRNA-NC):

Positive-sense strand is 5 '-UUCUCCGAACGUGUCACGU-3 ' (SEQ ID NO.8),

Antisense strand is 5 '-ACGUGACACGUUCGGAGAA-3 ' (SEQ ID NO.9)；

SiRNA1-LINC00511:

Positive-sense strand is 5 '-AAAGAGAAGACUCAGAUUCAA-3 ' (SEQ ID NO.10),

Antisense strand is 5 '-GAAUCUGAGUCUUCUCUUUCU-3 ' (SEQ ID NO.11)；

SiRNA2-LINC00511:

Positive-sense strand is 5 '-AUAACUCAAAAACAAACAGUC-3 ' (SEQ ID NO.12),

Antisense strand is 5 '-CUGUUUGUUUUUGAGUUAUUU-3 ' (SEQ ID NO.13)；

SiRNA3-LINC00511:

Positive-sense strand is 5 '-AAUUAGCAAGUCAAAACCCCC-3 ' (SEQ ID NO.14),

Antisense strand is 5 '-GGGUUUUGACUUGCUAAUUUA-3 ' (SEQ ID NO.15)

Cell is pressed 2 × 10⁵/ hole is inoculated into six porocyte culture plates, in 37 DEG C, 5%CO₂Cell culture in incubator 24h；

In DMEM culture medium without double antibody, containing 10%FBS, transfection (is purchased from according to lipofectamine 2000 Invitrogen company) specification transfection.

Experiment is divided into blank control group (HepG2) negative control group (siRNA-NC) and experimental group (20nM) (siRNA1- LINC00511, siRNA2-LINC00511, siRNA3-LINC00511), wherein negative control group siRNA and LINC00511 base For the sequence of cause without homology, concentration is the hole 20nM/, while being transfected respectively.

3, QPCR detects the transcriptional level of LINC00511 gene

The extraction of 3.1 cell total rnas

Specific steps are the same as embodiment 3.

3.2 reverse transcription steps are the same as embodiment 2.

3.3QPCR amplification step is the same as embodiment 2.

4, statistical method

Experiment is completed according to being repeated 3 times, and result data is indicated in a manner of mean+SD, Using SPSS18.0 statistical software come difference for statistical analysis, between interference LINC00511 gene expression panel and control group It is different to be examined using t, it is believed that there is statistical significance as P < 0.05.

5, result

As a result such as Fig. 3 is shown, compared to HepG2, transfection zero load siRNA-NC, siRNA1-LINC00511, siRNA3- LINC00511 group, siRNA2-LINC00511 group can significantly reduce the expression of LINC00511, and difference has statistical significance (P<0.05)。

5 CCK8 of embodiment detects cell proliferation experiment

1, cell culture and transfection procedure are the same as embodiment 4

2, CCK8 detects cell Proliferation

1) by the HepG2 cell inoculation of logarithmic proliferation phase in 96 orifice plates, every hole 2 × 10³A cell；

2) three groups of experiment point is blank control group, transfection siRNA-NC group respectively and transfects siRNA2-LINC00511, often Group sets 6 multiple holes；

3) respectively transfection 0h, for 24 hours, 10 hole μ l/ CCK8 reagents are added after 48h, 72h；

4) light absorption value of microplate reader detection A450 is used after 2h.

3, statistical method

Experiment is completed according to being repeated 3 times, using SPSS13.0 statistical software come for statistical analysis, the two it Between difference using t examine, it is believed that as P < 0.05 have statistical significance.

4, result

It is shown in Fig. 4 as the result is shown: blank control group transfects siRNA2-LINC00511 with unloaded group no significant difference The vitro growth rates of the obvious low control group of vitro growth rates of group, difference have statistical significance (P < 0.05), above-mentioned knot Fruit shows that the expression of LINC00511 can promote the growth of liver cancer cells.

6 soft-agar cloning of embodiment forms experiment

1, the cell of logarithmic growth phase is in 0.25% trypsin digestion, gently piping and druming makes unicellular outstanding Cell precipitation is collected by centrifugation in liquid.

2, it is resuspended with the DMEM complete medium containing 20% fetal calf serum, is counted after appropriate dilution, adjustment cell concentration is 5 ×10³A/ml.

3, the low melting point agar liquid glucose that two concentration are respectively 1.2% and 0.7% is prepared, after high pressure sterilization, maintains 40 In DEG C water-bath.

4,1.2% agarose and 2 × DMEM culture medium 1:1 mixing, are added the calf serum of 2 × antibiotic and 20%, It takes in 3ml mixed liquor injection diameter 6cm plate and places 5min cooled and solidified, be placed in CO as bottom-layer agar₂It is spare in incubator.

5, in sterile test tube 1:1 mixing 0.7% agarose and 2 × DMEM culture medium, then into pipe be added 0.2ml it is dense Degree is 5 × 10³A/ml's stablizes infection cell suspension, mixes well, injects in above-mentioned plate, gradually forms double agar layers, often A experimental group repeats 4 samples.

6, after top-layer agar solidification, 37 DEG C of 5%CO are placed in₂It is cultivated in incubator, every 3 days plus culture medium 1.5ml.

7, culture dish is taken out after cultivating 14 days, dyes 90min with the gentian violet that 1ml concentration is 0.005%.Plate is placed It is observed under inverted microscope, every group of cell randomly selects 10 low-power fields, the number of cell clones that technology is formed under mirror.

8, result

As a result as shown in figure 5, compared with the control group, transfecting the groups of cells single cell clone colony of siRNA2-LINC00511 Forming number significantly reduces.

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 LISTING

<110>Wang Dongguo

<120>a kind of biomarker for liver cancer diagnosis and treatment

<160> 15

<170> PatentIn version 3.5

<210> 1

<211> 2265

<212> DNA

<213>source of people

<400> 1

aaggggcgat gcgccccgga gggggaggga cgcaaggggc gactactgtt acctcgcttg 60

gagataccaa ggccggtcgg accccccgct ggccgaatgt ggggagcgag cagatccagc 120

ctgtttggac gtggtgagga cccagccgtg gggtttaagg ccaagttagc ctctcccttc 180

actgattaca agagccattg ttacaggtgc tcacatgcca ggagaaataa gctggtgatt 240

tatgtggcac agcaccatcg atcgacctac aaaggaagct tcagtgcagc cgtgttgcta 300

tacagacctc tctcaccaag gcgtcatgaa ctgaagggct gggagaggcc tcgtggagga 360

ggaagagttt cgctcagctg cttgggtgtg ggactgaatg tggttccaga actggtcgct 420

gccttcaccc gctgactctg gtctcggcga tgacaagccc atggaacaga cttacggcgt 480

atgacctcat cggagacttg tacatggggc caacccccat tttcccacag gaaacccaca 540

ctctaattaa gggctgaccc tgaacttgaa tctgagtctt ctctttcttc aacagagagc 600

catctctcca gcccagctgg caaggagaca ggtgatgtta caggaaaaaa tacacagctt 660

gtgcccttgg aattaccaga gttcaaagcc tgttcagcat ctacccacca tgtgacccgg 720

gtcgaattct tcagtttctc taagcccatt ttctcatctg taagatgggg aagtgtaatc 780

agttctgaac ccacatatac ctcactgggc ttgaacaggc ctgagcaggc atgtggggct 840

ttactagcta gccatccttt ctgccgctct ccctcattct ccttcaagcc ccccaaagtc 900

cttggtggag gaggaagtgg agacagaacg tttggtgttg ggtgaggtac gtgaggaggc 960

tgatttcaga acctccagga aacaaagctt tagtgttccg agggcagtgc ccgcctgtgc 1020

ctcctcctca cagggggtag taggagtggg gtggggccga ggcatttccc agcacagctc 1080

aatcacatct cctgcgcctg cccttgtgac ctgctcactg gaaaggaaga aatgaccgag 1140

ggagaaggga gggcatttga ggggagaaga agaggatgga gaaaaaaact ggatcatctc 1200

gtgacatgga aagaaaggtc tgagctctga cgatcccctc acgtgattat aggcatgagg 1260

actcactctt cgaattgttt gaaaattgaa tggactggct cacaccttct gaaatttctt 1320

ggaagctctg gagtaactat tccactaaag aacagcgtgt ttttggctga aatctgtgac 1380

tgtttgtttt tgagttattt taatctctgt gtcaagcatg aaaacctcag cagcctggga 1440

tttataatta gatcccactg gcatgccctc tcacctctca aacatgccac agtgctttgg 1500

ctctctcctt gtgcaaacct ccctttgata tttttggata tcgataccaa gtgtatctgc 1560

atattttaac caactgatat gcatatcctg tgatctgagg cattcgctca atgtggggca 1620

tcaggccagc tgtgaacttc ctttaaaatt tattttgtgt caaatgattc cctaatgcta 1680

tagctccctt gaactgctag atttatgaaa attctgcaca acacatagct cagagaaaaa 1740

aaaaaaagag tgttgcaaga ccagatatag gatatcattt gatttgttta aaacactctg 1800

tccagggggt tttgacttgc taatttaaaa ataattctaa aattaggtac agttgaccct 1860

tagtatccat gggggattgg ttccaggatg ccctgtggat accagaatcc aaggatgctc 1920

aagttcctga cataaatggc ttagtgtttg catataacct actcccctct tcctatatac 1980

tttaaataat ctctagatta cttataatgc ctaacacaac gcctgtgcat cacttcattc 2040

atgtggattc aatgtagtac ttggtgtgaa gaaaatttaa gtcttgcttt ttggaacttt 2100

gtggattttt ttttctggaa tgtttttgat ctgcagttgg ttgaatccac aggtgcagaa 2160

ctatggacac agggggctga ctatgtatat ttatatccac ttgcaaaatt aagaataaaa 2220

ataatgtgca tcagcaggtg cagattctga tgctcggtgg acagt 2265

<210> 2

<211> 1716

<212> DNA

<213>source of people

<400> 2

agggcgcgca ggcggcgcgg gtgcgcggtg cggcgctggt atccagagga cgcggtcacc 60

gcctctggca tttgtcgttc tgcgcttctc cgcaaggacc ctctgttagg caggcgccca 120

ccgtaagcct cccgggcctt gtgaacctgc aaacccaagt ctgagagacg atccgccttc 180

agcgctttcc agcttggcag agaggctttc ccggcgggga tctttggttg gcgctggcga 240

tgcgcgggga agaaaggcga ggagcggcgt ccaggctggg tgatgtccca gcacgagtag 300

gcgggatgcg ctcgcttggt cctccgggcg cccggtccct gcccgcgtcg cgcgcccacc 360

cctggggacg agaaggcggc cgcctgagga cccccgcccg cgacctccgc gagtctggag 420

cgcagaggac agggtctggc tgctctttgg ccttggatgg aaagtgggga attgggtggg 480

gggctgcgga ccccttaacg tggattactt ggtgtgtatc agctgggctc agaagaccca 540

cgacctcttc tccatccgtg gattgatttg ttctgcttaa cagctgggtc gccaagctgg 600

aggtattttt ccctctccac cctggtcttc tcctgtaacg tgtggccgcc ttttccagca 660

cggcctcctg ccttcctggt gcactttttg gagaacgtgg tggaatcaga ggtttctggc 720

tgactcggtg ggtgctttga accaggaaag gacaagaaag aggttggagt acagtggcat 780

gatctcagct cactgcagcc tcgacttcct gggctcaaga aatcctcctg tctctgcctt 840

ccgagtagct gtgatcacag ttgtatgcca tcatgcccag accagatttc gccatattgc 900

tcaggatggt cttgaactcc tgggctcaag tgatccgcct gccttggcct cccagaatgc 960

caggattaca gatgtgagcc actgtgcctg gcctgaagtc tagtaatttt taaaactttg 1020

catgttattc tgctcatgag ccaagttcgg gtgcctgtgg aacagctata ttcctcaaac 1080

tagatatgca ccccagtccc tggggattct gtcaaaaggc agattcacat tcagcaggtc 1140

tagggtgggg ggcctgacat tcggcacgtc tcacaagctc ccaggcgatg ctgatggtgc 1200

tggtccctgg acgccctctt gagtagtaag gaacaaaagg actctcccct gttcttttca 1260

gctacgtagc ccagtggttc tccacttagg cttcacactg gaatcatcca gggagtttta 1320

aactatagtg atgcctggcg gtgtcttact cgcagagctt ctggtttgat tggtctaggt 1380

tgcagcctgg gcaaggagat ttaaaaactt ctcagatggg ccaggcatgg tggctcaagc 1440

ctgtaatccc agcactttgg gaggcagagg tgggcggatc acgaggtcag gagatcgaga 1500

ccatcctggc tagcacggcg aaaccctgtc tctactaaaa atacaaaaaa ttagctgggt 1560

gtggtggcgt gtgcctgtgg tcccagctgc tcgggaggct gaggcagaag aatggcctga 1620

acccgggagg cggagcttgc agacagccga tcgtgccact gcactccagc ctgggcgaca 1680

gagcgagact ccgtctcaaa aaaaaaaaaa aaaaaa 1716

<210> 3

<211> 1696

<212> DNA

<213>source of people

<400> 3

tggggaattg ggtggggggc tgcggacccc ttaacgtgga ttacttggtg tgtatcagct 60

gggctcagaa gacccacgac ctcttctcca tccgtggatt gatttgttct gcttaacagc 120

tgggtcgcca agctggaggt ggaatcagag gtttctggct gactcggtgg gtgctttgaa 180

ccaggaaagg acaagaaaga ggatgggaag gactgatcca cattcccacc aggaagttta 240

gcagaacccc cgcgtgccac ctggacccct tggaaggacc tggctcaggc tggaccacct 300

cttgagaggc aggagctctg gatttgatca agaattcttt gctgagcatg gtgcctcatg 360

cctataatcc caacactttg ggaggccagt gtgggaggat ctcttgagcc caggagttca 420

agactagcct gggcaacaca gagagacccc atctctaaaa taataataat aataaaataa 480

aaaattagca gggcatggtg gcatgtgcct gtagtcccag ctacccagga ggctgaggca 540

agaggatggc tggagcctgg gatgttgagg ctgcaatgaa ctgtgattac cccactgcac 600

tccagcctgg gcaaaagagc gagagaccct gtctcaaata ataataataa taataatctt 660

attttggaga ataaagagac ctctggattt gaggtgccat ttgggtagaa agaaaagacg 720

tttacaccga gaaatagtct gtgttgccct gaaggagcag agggatgcat cgctggaggt 780

gacctacagt tgaagaagac tcattatgac agaccttgtc cttcttcctt gtggaaagtg 840

tttcctctgc tgctactgct catgagactc ttccccctcc ctgtcccagg gaaccaaagg 900

gctttctacc acaccctttc ttgccccccg cctcccatgt ctgctgtgcc tttgtactca 960

gcaattcttg tttgctccat tatcttccag ccggatacag agtgaatagt taaccacact 1020

taggtcaaat aggatctaaa tttttgttcc tgctccgtgt aaagaggcca gtgtttgtgt 1080

gttgcaagca gccttggaat agtaactctt ctcatttgtt tgggatctgg ccaccaagtt 1140

ccagaatgat acacggatca gtgcagaagt tcatcaggct ctcggacctt agggctgttg 1200

gagaaggctt cagcagcaga actgatggtg aaggctcgtg ttctccatcc tcaactttct 1260

ttgcttcgat catacacaag aatacatttg gaagggcaaa aaatgaacac tgtcgttcat 1320

tgcagccgtg ttttgtgaca cagatgcaca gtctgctgtg aagaccttct ctcaagtggc 1380

atttgggagt ccatgccaga tcatggtgct tcatgagaga ctgacagcta tcaggggttg 1440

tggcacttag tgaggactct cctcccccag tgtgtgctga tgacacatac acacctgaca 1500

atagcttgag tcttctctgt tccttttact ctgtagccaa catacacatg atttaaaacc 1560

ctttctaaat atctatcatg gttcatcctt gtccaaatgc agagtcagag ctatttgtac 1620

ttcattatta tttccaaggc gaatagttgg ctttcttttt gcaaaaataa ttaaagtttt 1680

tgtatgttgc agttgc 1696

<210> 4

<211> 19

<212> DNA

<213>artificial sequence

<400> 4

gctcaagttc ctgacataa 19

<210> 5

<211> 18

<212> DNA

<213>artificial sequence

<400> 5

gcgttgtgtt aggcatta 18

<210> 6

<211> 22

<212> DNA

<213>artificial sequence

<400> 6

ccgggaaact gtggcgtgat gg 22

<210> 7

<211> 25

<212> DNA

<213>artificial sequence

<400> 7

aggtggagga gtgggtgtcg ctgtt 25

<210> 8

<211> 19

<212> RNA

<213>artificial sequence

<400> 8

uucuccgaac gugucacgu 19

<210> 9

<211> 19

<212> RNA

<213>artificial sequence

<400> 9

acgugacacg uucggagaa 19

<210> 10

<211> 21

<212> RNA

<213>artificial sequence

<400> 10

aaagagaaga cucagauuca a 21

<210> 11

<211> 21

<212> RNA

<213>artificial sequence

<400> 11

gaaucugagu cuucucuuuc u 21

<210> 12

<211> 21

<212> RNA

<213>artificial sequence

<400> 12

auaacucaaa aacaaacagu c 21

<210> 13

<211> 21

<212> RNA

<213>artificial sequence

<400> 13

cuguuuguuu uugaguuauu u 21

<210> 14

<211> 21

<212> RNA

<213>artificial sequence

<400> 14

aauuagcaag ucaaaacccc c 21

<210> 15

<211> 21

<212> RNA

<213>artificial sequence

<400> 15

ggguuuugac uugcuaauuu a 21

Claims

1. detecting the purposes of the reagent of LINC00511 gene expression dose, which is characterized in that be used to prepare the production of diagnosing liver cancer Product.

2. purposes according to claim 1, which is characterized in that the LINC00511 expresses up-regulation in liver cancer patient.

3. purposes according to claim 1, which is characterized in that the product can by RT-PCR, real-time quantitative PCR, In situ hybridization, northern blotting, chip or high-flux sequence detection of platform LINC00511 expression.

4. purposes according to claim 1, which is characterized in that the product includes chip, preparation or kit.

The purposes of the inhibitor of 5.LINC00511 functional expression, which is characterized in that be used to prepare prevention or treat the medicine of liver cancer Compositions.

6. purposes according to claim 5, which is characterized in that described pharmaceutical composition further includes and the inhibitor compatibility Its other medicine class and pharmaceutically acceptable carrier and/or auxiliary material.