CN114525342A - Application of LINC02806 in diagnosis and treatment of hepatocellular carcinoma - Google Patents

Abstract

The invention discloses a biomarker for diagnosis and treatment of liver cancer, and particularly relates to a biomarker LINC 02806. The invention discovers that the differential expression of LINC02806 is related to the occurrence and development of liver cancer for the first time, and the expression of LINC02806 is up-regulated in liver cancer patients, thus prompting that the intervention of the gene expression of LINC02806 can become a new way for treating liver cancer. To further verify, it was found that the proliferation of liver cancer cells and the number of cell-forming clonal colonies could be significantly reduced by reducing the expression of LINC02806 by an inhibitor.

Description

Application of LINC02806 in diagnosis and treatment of hepatocellular carcinoma

Technical Field

The invention relates to the technical field of biological medicines, in particular to application of LINC02806 in diagnosis and treatment of hepatocellular carcinoma.

Background

Liver cancer is one of the leading causes of cancer death worldwide, and particularly in china, liver cancer has become one of the most common cancers due to the high infection rate of hepatitis viruses. Hepatocellular carcinoma is the most common type in primary liver cancers. Hepatitis b virus and hepatitis c virus infection remains a major cause of hepatocellular carcinoma. Liver cancer patients have poor prognosis and have an increasing incidence worldwide due to late stage diagnosis and limited treatment options. Median survival in untreated patients with advanced liver cancer is only 7.1 months. Even after treatment, tumor recurrence is a problem. For example, recurrence occurs in approximately 70% of patients 5 years post-surgery, reducing patient survival. Therefore, there is an urgent need to elucidate the molecular pathogenesis of liver cancer progression and determine the optimal diagnosis and treatment strategies, thereby contributing to the development of new diagnostic markers and therapeutic targets and improving the treatment efficiency and prognosis clinically.

lncRNA are RNA molecules of non-open reading frames >200 nucleotides in length. Although lncrnas have no protein coding ability, they are important in epigenetics and in regulating gene expression. Numerous studies have demonstrated that lncRNA is involved in brain development, embryonic development, tissue differentiation and organogenesis. In recent years, lncRNA has been seen as a key role in a variety of human diseases including liver cancer. For example, lncRNA MAGI2-AS3 may have a protective effect on liver cancer, lncRNA-HIS is thought to promote the proliferation and metastasis of HCC, and lncRNA-DUXAP10 plays a role in inhibiting the proliferation of liver cancer cells through microRNA-1914. This means that lncRNA may be a potentially effective biomarker for diagnosis and treatment of liver cancer.

At present, the regulation and control molecular mechanism of lncRNA in liver cancer is still in an exploration stage, the action of a plurality of unknown lncRNA in liver cancer needs to be researched, the method has important effects on diagnosis, prognosis and prediction of liver cancer, and an effective treatment target is provided for targeted treatment of liver cancer.

Disclosure of Invention

In order to make up for the defects of the prior art, the invention provides a product for diagnosing early liver cancer, so that a patient can be treated at an early stage, and the survival rate and the life quality are improved.

The second purpose of the invention is to provide a treatment means and a pharmaceutical composition for realizing precise molecular therapy of liver cancer.

The third object of the present invention is to provide a method for screening a substance to be screened for preventing or treating liver cancer.

In order to achieve the purpose, the invention adopts the following technical scheme:

the invention provides an application of a reagent for detecting LINC02806 in preparing a product for diagnosing liver cancer.

The reagent comprises a reagent for detecting the expression level of LINC02806 by using RT-PCR, real-time quantitative PCR, in-situ hybridization, northern blotting, a chip or a high-throughput sequencing platform.

Further, the reagent for detecting the LINC02806 by using real-time quantitative PCR at least comprises a pair of primers for specifically amplifying the LINC 02806.

Further, the reagent comprises:

a probe that specifically recognizes LINC 02806; or

And (3) primers for specifically amplifying LINC 02806.

Further, the primer sequence of the specific amplification LINC02806 is shown as SEQ ID NO.1 and SEQ ID NO. 2.

The invention provides a product for diagnosing liver cancer, which comprises a reagent for detecting the expression level of LINC02806 in a sample.

The product comprises a chip, a preparation or a kit. Wherein the chip comprises a solid phase carrier and oligonucleotide probes fixed on the solid phase carrier, and the oligonucleotide probes comprise oligonucleotide probes aiming at LINC02806 and used for detecting the transcription level of LINC 02806; the kit comprises a primer or a chip for detecting the transcription level of LINC 02806.

The solid phase carrier can be made of various materials commonly used in the field of gene chip, such as but not limited to nylon membrane, glass or silicon slice modified by active group (such as aldehyde group, amino group, etc.), unmodified glass slice, plastic slice, etc.

The gene detection kit or the gene chip can be used for detecting the expression levels of a plurality of genes (for example, a plurality of genes related to liver cancer) including the LINC02806 gene, and can simultaneously detect a plurality of markers of the liver cancer, thereby greatly improving the accuracy of liver cancer diagnosis.

Further, the reagent for detecting the level of LINC02806 in the sample comprises:

a probe that specifically recognizes LINC 02806; or

And (3) primers for specifically amplifying LINC 02806.

Further, the primer sequence for specifically amplifying LINC02806 is shown as SEQ ID NO.1 and SEQ ID NO. 2.

The invention provides application of LINC02806 in preparation of a pharmaceutical composition for treating liver cancer.

Further, the pharmaceutical composition comprises an inhibitor of LINC02806, and/or other medicaments compatible with the inhibitor and a pharmaceutically acceptable carrier and/or adjuvant, wherein the inhibitor can inhibit the expression of LINC02806 or substances related to the upstream or downstream pathway of LINC 02806.

Further, the inhibitor is an siRNA directed against LINC 02806.

When the invention is used for screening effective siRNA sequences, the optimal effective segment is found out through a large amount of comparison analysis. In the specific implementation mode of the invention, the inventor designs and synthesizes a plurality of siRNA sequences, and verifies the siRNA sequences by respectively transfecting a liver cancer cell line with a transfection reagent, so that the result detects interference molecules with better interference effect, the interference molecules have the sequences shown in SEQ ID NO.9 and SEQ ID NO.10 respectively, and further the result of cell level experiments proves that the inhibition efficiency is very high for cell experiments.

The nucleic acid inhibitor of the present invention, such as siRNA, can be chemically synthesized or can be prepared by transcribing an expression cassette in a recombinant nucleic acid construct into single-stranded RNA. Nucleic acid inhibitors, such as siRNA, can be delivered into cells by using appropriate transfection reagents, or can also be delivered into cells using a variety of techniques known in the art.

One skilled in the art will recognize that the utility of the present invention is not limited to quantifying gene expression of any particular variant of the marker genes of the present invention. In a specific embodiment, as a non-limiting example, the marker gene LINC02806 has the sequence shown by the LINC02806 gene (NC _000001.11) in the current international public nucleic acid sequence database, GeneBank.

In the present invention, the term "probe" refers to a molecule that is capable of binding to a specific sequence or subsequence or other portion of another molecule. Unless otherwise indicated, the term "probe" generally refers to a polynucleotide probe that is capable of binding to another polynucleotide (often referred to as a "target polynucleotide") by complementary base pairing. Depending on the stringency of the hybridization conditions, a probe can bind to a target polynucleotide that lacks complete sequence complementarity to the probe. The probe may be directly or indirectly labeled, and includes within its scope a primer. Hybridization modalities, including, but not limited to: solution phase, solid phase, mixed phase or in situ hybridization assays.

In the present invention, pharmaceutically acceptable carriers include (but are not limited to): diluents, excipients such as lactose, sodium chloride, glucose, urea, starch, water, etc., fillers such as starch, sucrose, etc.; binders such as simple syrup, glucose solution, starch solution, cellulose derivatives, alginates, gelatin, and polyvinylpyrrolidone; humectants such as glycerol; disintegrating agents such as dry starch, sodium alginate, laminarin powder, agar powder, calcium carbonate and sodium bicarbonate; absorption accelerators quaternary ammonium compounds, sodium lauryl sulfate, and the like; surfactants such as polyoxyethylene sorbitan fatty acid esters, sodium lauryl sulfate, glyceryl monostearate, cetyl alcohol, etc.; humectants such as glycerin, starch, etc.; adsorption carriers such as starch, lactose, bentonite, silica gel, kaolin, and bentonite, etc.; lubricants such as talc, calcium and magnesium stearate, polyethylene glycol, boric acid powder, and the like.

In the present invention, the pharmaceutical composition may be prepared using various additives, such as buffers, stabilizers, bacteriostats, isotonizing agents, chelating agents, pH controlling agents, and surfactants.

The pharmaceutical compositions of the present invention may be administered orally, parenterally, by inhalation spray, topically, rectally, nasally, buccally, vaginally or via an implanted reservoir. Oral administration or injection administration is preferred. The pharmaceutical compositions of the present invention may contain any of the usual non-toxic pharmaceutically acceptable carriers, adjuvants or vehicles. In some cases, pharmaceutically acceptable acids, bases or buffers may be used to adjust the pH of the formulation to improve the stability of the formulated compound or its dosage form in which it is administered. The term parenteral as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, Ju's, intralesional, and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may be administered to a subject by any route as long as the target tissue is reached.

The pharmaceutical compositions of the present invention may be administered orally in any oral dosage form including, but not limited to, capsules, tablets, emulsions and aqueous suspensions, dispersions and solutions. For oral tablets, carriers that are commonly used include lactose and corn starch. Lubricating agents such as magnesium stearate are also typically added. For oral administration in capsule form, suitable diluents include lactose and anhydrous corn starch. When aqueous suspensions and/or emulsions are administered orally, the active ingredient may be suspended or dissolved in the oil phase and combined with emulsifying and/or suspending agents. If desired, sweetening and/or flavouring and/or colouring agents may be added. Dosage unit formulations for oral administration may be microencapsulated, as appropriate. The formulations may also be prepared to provide extended or sustained release, for example, by coating or embedding the particulate material in a polymer, wax, or the like. The pharmaceutical composition can be used for reducing endogenous LINC02806 overexpression and reducing the expression of LINC02806, so that the liver cancer caused by the up-regulation of the LINC02806 expression is treated.

In the present invention, the compound that inhibits expression of LINC02806 can be administered to a subject as a naked RNA along with a delivery agent as a nucleic acid (e.g., a recombinant plasmid or viral vector) comprising a sequence that inhibits expression of LINC 02806. The delivery agent may be a lipophilic agent, a polycation, a liposome, or the like.

The pharmaceutical compositions of the present invention may further comprise one or more anti-cancer agents. In a specific embodiment, the pharmaceutical composition comprises at least one compound that inhibits the expression of the LINC02806 gene and at least one chemotherapeutic agent. Chemotherapeutic agents for use in the present invention include, but are not limited to: microtubule activators, alkylating agents, antineoplastic antimetabolites, platinum-based compounds, DNA-alkylating agents, antineoplastic antibiotic agents, antimetabolites, tubulin stabilizing agents, tubulin destabilizing agents, hormone antagonists, topoisomerase inhibitors, protein kinase inhibitors, HMG-COA inhibitors, CDK inhibitors, cyclin inhibitors, caspase inhibitors, metalloproteinase inhibitors, antisense nucleic acids, triple helix DNA, nucleic acid aptamers, and molecularly modified viral, bacterial and exotoxin agents.

The medicament of the invention can also be used in combination with other medicaments for treating liver cancer, and other therapeutic compounds can be simultaneously administered with the main active ingredients, even in the same composition. Other therapeutic compounds may also be administered alone in a composition or dosage form different from the main active ingredient. Some of the doses of the main ingredient may be administered simultaneously with other therapeutic compounds, while other doses may be administered separately. The dosage of the pharmaceutical composition of the present invention can be adjusted during the course of treatment depending on the severity of symptoms, the frequency of relapse, and the physiological response of the treatment regimen.

The pharmaceutical composition of the present invention may be a pharmaceutical composition for topical administration, and may be formulated as an ointment, cream, suspension, lotion, powder, solution, paste, gel, spray, aerosol or oil.

The pharmaceutical composition of the present invention may be administered in a pharmaceutically effective amount, and the term "pharmaceutically effective amount" of the present invention means an amount sufficient to treat or prevent a disease at a reasonable benefit/risk ratio applicable to medical treatment or prevention, and the effective dosage level may be determined according to factors including the severity of the disease, the activity of the drug, the age, body weight, health, sex, patient's sensitivity to the drug, administration time of the composition of the present invention to be used, administration route and excretion ratio, treatment time, factors of the drug to be used in combination or concomitantly with the composition of the present invention to be used, and other factors known in the medical field. The pharmaceutical composition of the present invention may be administered as a single therapeutic agent or in combination with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. In addition, administration may be performed in a single or multiple doses. It is important to consider all of the above elements and to administer them in an amount that achieves the maximum effect with the minimum amount of side effects.

The term "treatment" refers to not the purpose of curing, but rather slowing (reducing) the targeted pathological condition or disorder or preventing relapse. If a patient is successfully "treated" after receiving a therapeutically effective amount of a therapeutic agent, the patient exhibits observable and/or measurable reduction or disappearance of signs and symptoms of one or more particular diseases. For example, a significant reduction in the number of cancer cells or disappearance of cancer cells, reduction in tumor size; inhibit (i.e., slow to some extent, and preferably stop) tumor metastasis; inhibit tumor growth to some extent; increasing the time to reduce to some extent and/or alleviate one or more symptoms associated with a particular cancer; reduced morbidity and mortality, and improved quality of life. The reduction in signs or symptoms of the disease can be perceived by the patient. Treatment may achieve a complete response-defined as the disappearance of all signs of cancer, or a partial response-a reduction in tumor size, preferably by a proportion of more than 50%, more preferably 75%. Patients are also considered treated if they experience stable disease.

In the present invention, the term "sample" includes, but is not limited to, cells, tissues, organs, body fluids (blood, lymph, etc.), digestive juices, expectorations, alveolar bronchial washes, urine, feces, etc. Preferably, the sample is tissue or blood.

The invention also provides application of LINC02806 in screening substances to be screened for preventing or treating liver cancer.

Further, the step of screening a substance to be screened for preventing or treating liver cancer comprises:

treating a system expressing or containing the LINC02806 gene by a substance to be screened; and

detecting the expression of the LINC02806 gene in the system;

wherein, if the substance to be screened can reduce the expression or activity of the LINC02806 gene (preferably significantly reduce, such as by more than 20%, preferably by more than 50%, more preferably by more than 80%), it indicates that the substance to be screened is a substance to be screened for preventing or treating liver cancer.

Further, the systems described above include (but are not limited to): a cell system, a subcellular system, a solution system, a tissue system, an organ system, or an animal system.

Compared with the prior art, the invention provides a product for diagnosing early liver cancer, so that a patient can be treated at an early stage, and the survival rate and life treatment are further improved. The invention also provides a treatment means and a pharmaceutical composition, which can realize the precise molecular treatment of the liver cancer. The invention provides a method for screening substances to be screened for preventing or treating liver cancer, which has important effects on diagnosis, prognosis and prediction of liver cancer.

Drawings

FIG. 1 is a graph showing the detection of the expression of LINC02806 in liver cancer patients using QPCR;

FIG. 2 is a graph showing the detection of the expression of LINC02806 in liver cancer cells by QPCR;

FIG. 3 is a graph showing the effect of QPCR on the expression of LINC02806 in HepG2 hepatoma cells by transfected siRNA;

FIG. 4 is a graph showing the effect of LINC02806 on HepG2 liver cancer cell proliferation measured by CCK 8.

Detailed Description

The present invention will be described in further detail with reference to the accompanying drawings and examples. The following examples are intended to illustrate the invention only and are not intended to limit the scope of the invention. Experimental procedures without specific conditions noted in the examples, generally following conventional conditions, such as Sambrook et al, molecular cloning: the conditions described in the laboratory Manual (New York: Cold Spring harbor laboratory Press,1989), or according to the manufacturer's recommendations.

Example 1: screening of Gene markers associated with liver cancer

1. Sample collection

Cancer tissues and tissues adjacent to the cancer were collected from 5 patients with liver cancer, and the patients gave their informed consent, and all of the above specimens were obtained with the consent of the tissue ethics committee.

2. Preparation of RNA samples

Tissue RNA extraction was performed using a tissue RNA extraction kit from QIAGEN, and the procedures were performed according to the specific procedures described in the specification.

3. Mass analysis of RNA samples

The concentration and purity of the extracted RNA were determined using Nanodrop2000, RNA integrity was determined by agarose gel electrophoresis, and RIN was determined by Agilent 2100. The concentration is more than or equal to 200 ng/mul, and the OD260/280 is between 1.8 and 2.2.

4. Removal of rRNA

Ribosomal RNA was removed from total RNA using Ribo-Zero kit.

5. Construction of cDNA library

The construction of cDNA library was carried out using the Truseq RNA sample Prep Kit from Illumina, the detailed procedures were as described in the specification.

6. Sequencing on machine

The cDNA library was sequenced using the Hiseq4000 sequencing platform, the specific procedures were as described in the specification.

7. High throughput transcriptome sequencing data analysis

Performing bioinformatics analysis on original count data by adopting R-packet DESeq2, deleting non-detectable lncRNA, and performing differential expression lncRNA screening standard: FDR <0.05, abs (log2FC) > 2.

7. Results

RNA-seq results show that the expression level of LINC02806 in liver cancer tissues is obviously higher than that in paracancer tissues.

Example 2: QPCR sequencing verification of differential expression of LINC02806 gene

1. Large sample QPCR validation was performed on differential LINC02806 gene expression. 50 samples of liver cancer tissue and paracancerous tissue were collected in the same manner as in example 1.

2. The RNA extraction procedure was as in example 1.

3. Reverse transcription:

a20-mu-l reaction system is adopted, 1 mu g of total RNA is taken from each sample as template RNA, and the following components are respectively added into a PCR tube: DEPC water, premixed reagent, template RNA. 15min at 37 ℃, 5min at 50 ℃, 5min at 98 ℃ and maintaining at 4 ℃.

(3) QPCR amplification assay

Primers were designed based on the sequences of LINC02806 gene and housekeeping gene GAPDH, and the primer sequences were synthesized by Shanghai. The designed primer sequences are shown below:

the primer sequence of the LINC02806 gene is as follows:

a forward primer: 5'-CACGATCCACATCTCAGGGG-3' (SEQ ID NO.1)

Reverse primer: 5'-TCAACACACTCGGAAGAGGC-3' (SEQ ID NO.2)

The primer sequence of housekeeping gene GAPDH is as follows:

a forward primer: 5'-GAAAGCCTGCCGGTGACTAA-3' (SEQ ID NO.3)

Reverse primer: 5'-GCCCAATACGACCAAATCAGAG-3' (SEQ ID NO.4)

Prepare 10 μ l of the following reaction system: SYBR Green polymerase chain reaction system 5. mu.l, forward and reverse primers (5. mu.M) 2. mu.l each, and template cDNA 1. mu.l. All operations were performed on ice. Each sample was provided with 3 parallel channels and all amplification reactions were repeated three more times to ensure the reliability of the results. The amplification procedure was: 60s at 95 deg.C, (10 s at 95 deg.C, 30s at 60 deg.C). times.40 cycles.

SYBR Green is used as a fluorescent marker, PCR reaction is carried out on a 7500Fast fluorescent real-time quantitative PCR instrument, a target band is determined through melting curve analysis and electrophoresis, and relative quantification is carried out through a delta CT method.

3. Statistical method

The experiments were performed in 3 replicates, the results were represented as mean ± sd, and were statistically analyzed using SPSS18.0 statistical software, with the difference between the two using the t-test, and considered statistically significant when P < 0.05.

4. Results

As shown in FIG. 1, the expression of LINC02806 gene was up-regulated in liver cancer tissue compared with that in the tissue adjacent to the cancer, and the difference was statistically significant (P <0.05), which is consistent with the result of RNA-sep.

Example 3: differential expression of LINC02806 Gene in liver cancer cell lines

1. Cell culture

Human hepatoma cell lines HepG2, Huh7 and normal liver cell line HL-7702 were cultured in DMEM medium containing 10% fetal bovine serum and 1% P/S at 37 deg.C, 5% CO2 and 90% relative humidity. The solution was changed 1 time for 2-3 days and passaged by conventional digestion with 0.25% trypsin containing EDTA.

2. Extraction of RNA

The RNA in the cells was extracted using Qiagen's cell RNA extraction kit, and the experimental procedures were performed according to the instructions.

3. The reverse transcription was performed as in example 2

4. Statistical method

The experiments were performed in 3 replicates, the results were represented as mean ± sd, and were statistically analyzed using SPSS18.0 statistical software, with the difference between the two using the t-test, and considered statistically significant when P < 0.05.

5. Results

As shown in FIG. 2, compared with the normal liver cell line, the expression of the LINC02806 gene is up-regulated in liver cancer cells HepG2 and Huh7, and the difference is statistically significant (P <0.05), which is consistent with the result of RNA-sep.

Example 4 silencing of LINC02806 Gene

1. Cell culture

The same procedure as in example 3 was followed to culture a human hepatoma cell line HepG 2.

2. SiRNA design

Interfering RNAs were designed based on the sequence of LINC02806, dividing the experiment into four groups: a blank control group (HepG2), a negative control group (siRNA-NC) and an experimental group (siRNA1, siRNA2 and siRNA3), wherein the siRNA of the negative control group has no homology with the sequence of the LINC02806 gene. The designed siRNA sequence is shown as follows:

negative control siRNA sequence (siRNA-NC):

the sense strand is 5'-AAAUCAGAGAAUAAUCUAGGA-3' (SEQ ID NO.5),

the antisense strand is 5'-UUUAGUCUCUUAUUAGAUCCU-3' (SEQ ID NO. 6);

siRNA1：

the sense strand is 5'-UAUUACUCCCCUUAUUGCGGG-3' (SEQ ID NO.7),

the antisense strand is 5'-AUAAUGAGGGGAAUAACGCCC-3' (SEQ ID NO. 8);

siRNA2：

the sense strand is 5'-UCAAUGUGUUUUUGGAAUGCA-3' (SEQ ID NO.9),

the antisense strand is 5'-AGUUACACAAAAACCUUACGU-3' (SEQ ID NO. 10);

siRNA3：

the sense strand is 5'-AGAUACAGUAGGAAGAUGGCA-3' (SEQ ID NO.11),

the antisense strand is 5'-UCUAUGUCAUCCUUCUACCGU-3' (SEQ ID NO.12)

Inoculating cells into six-well cell culture plates at a speed of 2 × 105/well, and culturing the cells in a 5% CO2 incubator at a temperature of 37 ℃ for 24 h; transfection was performed in DMEM medium without double antibody containing 10% FBS according to the instructions of lipofectin 2000 (purchased from Invitrogen).

3. QPCR detection of transcription level of LINC02806 gene

3.1 extraction of Total RNA from cells the procedure was the same as in example 3.

3.2 reverse transcription procedure as in example 2.

3.3QPCR amplification step as in example 2.

4. Statistical method

The experiments were performed in 3 replicates, the data were expressed as mean ± sd, statistically analyzed using SPSS18.0 statistical software, and the difference between the interfering LINC02806 gene expression group and the control group was considered statistically significant when P <0.05 using t-test.

5. Results

The results are shown in fig. 3, where LINC02806mRNA levels were reduced in the experimental group compared to transfected empty siRNA-NC, where the interference effect was most significant in the siRNA2 group, and the difference was statistically significant (P < 0.05).

Example 5CCK8 assay for cell proliferation

1. Cell culture and transfection procedures were as in example 4

2. CCK8 detection of cell proliferation

1) The HepG2 cells in the logarithmic proliferation phase were seeded in 96-well plates at 2X 103 cells per well; dividing the experiment into groups (control group, siRNA-NC group and siRNA2 group), wherein each group is provided with 3 multiple wells;

2) adding 10 mul/well CCK8 reagent after transfection for 0h, 24h, 48h and 72h respectively;

3) after 2h, the absorbance of A450 was measured using a microplate reader.

3. Statistical method

The experiments were performed in 3 replicates using SPSS13.0 statistical software for statistical analysis, and the differences between the two were considered statistically significant when P <0.05 using the t-test.

4. Results

The results shown in fig. 4 show that: the cell growth rate of the siRNA2 group is obviously lower than that of the control group, the difference has statistical significance (P <0.05), and the result shows that the LINC02806 knockout can inhibit the growth of the liver cancer cells.

The above description of the embodiments is only intended to illustrate the method of the invention and its core idea. It should be noted that, for those skilled in the art, without departing from the principle of the present invention, several improvements and modifications can be made to the present invention, and these improvements and modifications will also fall into the protection scope of the claims of the present invention.

Claims (10)

1. Application of a reagent for detecting LINC02806 in preparation of a product for diagnosing liver cancer.

2. Use according to claim 1, characterized in that: the reagent comprises:

a probe that specifically recognizes LINC 02806; or

And (3) primers for specifically amplifying LINC 02806.

3. Use according to claim 2, characterized in that: the primer sequence of the specific amplification LINC02806 is shown as SEQ ID NO.1 and SEQ ID NO. 2.

4. A product for diagnosing liver cancer, which is characterized in that: the product comprises an agent for detecting the level of LINC02806 expression in a sample.

5. The product of claim 4, wherein: the product comprises a chip, a preparation or a kit.

Application of LINC02806 in preparing a pharmaceutical composition for treating liver cancer.

7. Use according to claim 6, characterized in that: the pharmaceutical composition comprises an inhibitor of LINC 02806.

8. Use according to claim 7, characterized in that: the inhibitor is siRNA directed against LINC 02806.

Application of LINC02806 in screening substances to be screened for preventing or treating liver cancer.

10. Use according to claim 9, characterized in that: the step of screening the substance to be screened for preventing or treating liver cancer comprises the following steps:

treating a system expressing or containing the LINC02806 gene by a substance to be screened; and

detecting the expression of the LINC02806 gene in the system;

wherein, if the substance to be screened can reduce the expression or activity of the LINC02806 gene, the substance to be screened is the substance to be screened for preventing or treating liver cancer.

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