CN110951882A - Application of LINC01012 as gastric cancer diagnosis molecule and drug target gene - Google Patents

Application of LINC01012 as gastric cancer diagnosis molecule and drug target gene Download PDF

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CN110951882A
CN110951882A CN201911399582.4A CN201911399582A CN110951882A CN 110951882 A CN110951882 A CN 110951882A CN 201911399582 A CN201911399582 A CN 201911399582A CN 110951882 A CN110951882 A CN 110951882A
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gastric cancer
linc01012
coding rna
long
expression
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杨承刚
陈丽媛
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Qingdao Yangshen Biomedical Co Ltd
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Beijing Medintell Bioinformatic Technology Co Ltd
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    • C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
    • C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
    • C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
    • C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
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    • A61K31/7088—Compounds having three or more nucleosides or nucleotides
    • A61K31/713—Double-stranded nucleic acids or oligonucleotides
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
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    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K49/00—Preparations for testing in vivo
    • A61K49/0004—Screening or testing of compounds for diagnosis of disorders, assessment of conditions, e.g. renal clearance, gastric emptying, testing for diabetes, allergy, rheuma, pancreas functions
    • A61K49/0008—Screening agents using (non-human) animal models or transgenic animal models or chimeric hosts, e.g. Alzheimer disease animal model, transgenic model for heart failure
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    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
    • G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
    • G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
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    • C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/158—Expression markers
    • C—CHEMISTRY; METALLURGY
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    • C12Q2600/00—Oligonucleotides characterized by their use
    • C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA

Abstract

The invention discloses application of LINC01012 as a gastric cancer diagnosis molecule and a drug target gene. The tissue level verification proves that the expression of LINC01012 in the stomach cancer tissue is increased; in-vitro cytofunctional experiments prove that the gene expression in the gastric cancer cells is obviously reduced and the cell proliferation capacity is reduced after the transfection of the siRNA interfering with the LINC 01012. The research result of the invention provides a new idea and a new method for clinical diagnosis and treatment of gastric cancer.

Description

Application of LINC01012 as gastric cancer diagnosis molecule and drug target gene
Technical Field
The invention belongs to the field of biomedicine, and relates to application of LINC01012 as a gastric cancer diagnosis molecule and a drug target gene.
Background
Gastric Cancer (GC) is the second largest cancer worldwide leading to death. The rapid progression and metastasis of the disease results in a poor prognosis. In order to improve early diagnosis and targeted therapy of GC, understanding of its underlying molecular mechanisms and identifying new, effective biomarkers and therapeutic targets are urgently needed.
Long-chain non-coding RNAs (lncrnas) are RNA molecules with transcripts longer than 200nt, different from other small-molecule non-coding RNAs, which can regulate RNA metabolism, protein functional activity, histone modification and chromatin remodeling in a cis (in cis) or trans (in trans) action manner, and are widely involved in cellular biological processes at epigenetics, transcription and post-transcriptional levels. Non-coding RNA (ncrna) is RNA that cannot be translated into protein. Two main categories are distinguished: small RNAs (< 200NT) and lncRNAs. Small ncRNA, including small RNA, siRNA and DNA, has been demonstrated to play an important role in gastric carcinogenesis. In recent years, a number of studies have shown that lncRNAs are involved in the development of a variety of diseases, having a variety of functions in a wide range of biological processes, particularly in connection with the development of human cancer. The study showed that: in addition to these coding genes, which have been extensively studied, abnormal expression of long noncoding RNAs (lncRNAs) is also involved in tumor malignant progression. More and more researches prove that the lncRNAs not only play an important role in maintaining the normal physiological functions and the development process of the body, but also have close relation between the abnormal expression and the occurrence and the development of diseases, particularly the malignant development of tumors. One well-known pathogenesis of tumor involvement of oncogenic lncRNA is HOTAIR (HOX transcriptional antisense RNA), which is consistently up-regulated and is identified as a prognostic marker of patient outcome, e.g., affecting survival in a variety of human tumor patients.
Recently, many studies have shown that lncRNA is highly expressed in gastric cancer tissues. Therefore, there is an urgent need to find new lncRNAs and investigate their biological effects on gastric cancer in order to obtain potential therapeutic targets for diagnosis and gene therapy of gastric cancer.
Disclosure of Invention
The invention aims to provide a long-chain non-coding RNA marker for diagnosing and treating gastric cancer. Experiments prove that the expression level of LINC01012 in gastric cancer tissues is obviously higher than that in para-cancer tissues, so that LINC01012 can be used as a molecular marker for diagnosis and treatment of gastric cancer.
In order to test the purpose, the invention adopts the following technical scheme:
the invention provides an application of a reagent for detecting long-chain non-coding RNA expression in preparation of gastric cancer diagnosis products.
The long non-coding RNA of the invention is named LINC01012 in NCBI, and the gene ID: 100507173, the transcript sequence of LINC01012 includes the following: genbank accession number NR _038292.1 (having a length of 2785bp, corresponding DNA sequence shown in SEQ ID NO. 1); genbank accession number NR _038293.1 (having a length of 675bp, the corresponding DNA sequence is shown in SEQ ID NO. 2); genbank accession number NR-038294.1 (having a length of 753bp, the corresponding DNA sequence being shown in SEQ ID NO. 3).
Further, the reagent comprises a PCR amplification primer used for detecting the LINC01012 expression level by using SYBR Green, a TaqMan probe, a molecular beacon, a double-hybridization probe or a composite probe.
In a specific embodiment of the invention, the primer sequences are shown as SEQ ID NO.4 and SEQ ID NO. 5.
The invention provides a product for gastric cancer diagnosis, which comprises a reagent for detecting the expression level of LINC 01012.
Further, the reagent comprises SYBR Green, a TaqMan probe, a molecular beacon, a double-hybridization probe or a PCR amplification primer used for detecting the LINC01012 expression quantity by a composite probe.
In a specific embodiment of the invention, the primer sequences are shown as SEQ ID NO.4 and SEQ ID NO. 5.
Further, the aforementioned products include, but are not limited to, chips, kits, test strips, or high throughput sequencing platforms; the high-throughput sequencing platform is a special tool for diagnosing gastric cancer, and with the development of a high-throughput sequencing technology, the construction of an RNA expression profile of a person becomes very convenient work. By comparing the RNA expression profiles of patients with disease and normal populations, it is easy to identify which RNA abnormalities are associated with disease. Therefore, the knowledge that non-LINC 01012 abnormalities are associated with gastric cancer in high throughput sequencing is also included in the use of LINC01012 and is also within the scope of the present invention.
The kit comprises a reagent for detecting the expression quantity of LINC01012, the reagent comprises nucleic acid combined with LINC01012 or a DNA sequence thereof, and the nucleic acid comprises SYBR Green, a TaqMan probe, a molecular beacon, a double-hybridization probe or a PCR amplification primer used when a composite probe is used for detecting the expression quantity of LINC 01012.
The chip comprises a reagent for detecting the expression level of LINC01012, wherein the reagent comprises a nucleic acid combined with LINC01012 or a DNA sequence thereof, and the nucleic acid comprises a probe capable of detecting the expression level of LINC 01012.
The test paper comprises a reagent for detecting the expression level of LINC01012, the reagent comprises a nucleic acid combined with LINC01012 or a DNA sequence thereof, and the nucleic acid comprises a probe capable of detecting the expression level of LINC 01012.
The present invention provides a pharmaceutical composition for the treatment of gastric cancer comprising an agent that inhibits LINC 01012.
Further, the agent is not limited as long as it can inhibit the expression level of LINC01012 or inhibit the functional activity of LINC 01012.
The reagent comprises siRNA or shRNA of LINC 01012. In a specific embodiment of the invention, the siRNA sequence of LINC01012 is shown in SEQ ID NO.8 and SEQ ID NO. 9.
The pharmaceutical composition of the present invention may be administered alone or together with other drugs as a medicine. The other drug that can be administered together with the pharmaceutical composition of the present invention is not limited as long as it does not impair the effect of the therapeutic or prophylactic pharmaceutical composition of the present invention.
The pharmaceutical composition of the invention can be prepared into various dosage forms according to requirements. Including, but not limited to, tablets, solutions, granules, patches, ointments, capsules, aerosols or suppositories for transdermal, mucosal, nasal, buccal, sublingual or oral use.
The route of administration of the pharmaceutical composition of the present invention is not limited as long as it can exert the desired therapeutic or prophylactic effect, and includes, but is not limited to, intravenous, intraperitoneal, intraocular, intraarterial, intrapulmonary, oral, intravesicular, intramuscular, intratracheal, subcutaneous, transdermal, transpleural, topical, inhalation, transmucosal, cutaneous, gastrointestinal, intraarticular, intraventricular, rectal, vaginal, intracranial, intraurethral, intrahepatic. In some cases, the administration may be systemic. In some cases topical administration.
The dosage of the pharmaceutical composition of the present invention is not limited as long as the desired therapeutic effect or prophylactic effect is obtained, and can be appropriately determined depending on the symptoms, sex, age, and the like. The dose of the therapeutic or prophylactic pharmaceutical composition of the present invention can be determined using, for example, the therapeutic effect or prophylactic effect on a disease as an index.
The invention also provides the application of the long-chain non-coding RNA in preparing a medicament for treating gastric cancer.
The invention also provides the application of the reagent for inhibiting the long-chain non-coding RNA in preparing the medicine for treating gastric cancer.
Further, the reagent is as defined above.
The present invention also provides a method for diagnosing gastric cancer, comprising the steps of:
(1) obtaining a sample from a subject;
(2) detecting the expression level of LINC01012 in a sample from the subject;
(3) correlating the measured expression level of LINC01012 with the presence or absence of disease in the subject.
(4) If the expression level of LINC01012 is increased compared to the control, the subject is determined to have gastric cancer, or the subject is at high risk of having gastric cancer, or the gastric cancer patient is determined to have poor prognosis.
The present invention also provides a method for the treatment of gastric cancer, said method comprising reducing the expression level of LINC01012 or inhibiting the functional activity of LINC 01012.
The invention also provides a screening method of the gastric cancer drug, which can measure the effect of the tumor drug on improving tumor prognosis by measuring the expression level of LINC01012 after adding the test drug to gastric cancer cells or at a certain period after applying the test drug to gastric cancer model animals. More specifically, when the expression level of LINC01012 decreases or returns to normal levels after addition or administration of the test drug, the drug can be selected as a therapeutic drug for improving the prognosis of tumor.
In the context of the present invention, "diagnosing gastric cancer" includes determining whether a subject has gastric cancer, determining whether a subject is at risk of having gastric cancer, determining responsiveness of a gastric cancer patient to drug treatment, or determining a prognosis for a gastric cancer patient.
As used herein, "treatment" encompasses treatment-related diseases or disease states in a mammal, such as a human, having the associated disease or disorder, and includes:
(1) preventing the occurrence of a disease or condition in a mammal, particularly when the mammal is susceptible to said disease condition but has not been diagnosed as having such a disease condition;
(2) inhibiting a disease or disease state, i.e., preventing its occurrence; or
(3) Alleviating the disease or condition, i.e., causing regression of the disease or condition.
The term "treatment" generally refers to the treatment of a human or animal (e.g., as applied by a veterinarian) wherein some desired therapeutic effect is achieved, e.g., inhibiting the progression of a condition (including slowing the progression, stopping the progression), ameliorating the condition, and curing the condition. Treatment as a prophylactic measure (e.g., prophylaxis) is also included. The use of a patient who has not yet developed a condition but who is at risk of developing the condition is also encompassed by the term "treatment".
The invention has the advantages and beneficial effects that:
the invention discloses a molecular marker for diagnosing gastric cancer, which can be used for judging the early stage of gastric cancer occurrence and provides the survival rate of patients.
The therapeutic agent of the present invention comprising an agent that inhibits LINC01012 can be used as a novel therapeutic agent for gastric cancer.
Drawings
FIG. 1 shows a statistical chart of the detection of the inhibition of LINC01012 expression by QPCR.
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 study of differential expression of LncRNA
1. Study object
Surgical specimens (gastric cancer tissues and corresponding paracancerous normal tissues) of 50 primary gastric cancer patients subjected to gastric carcinoma radical surgery in tumor surgery in hospitals were collected. 30 men and 20 women in 50 patients; the patients are 38-79 years old, and the average age is 61 years old, and the patients have not received adjuvant treatment before operation. Immediately storing the tissue specimen at-80 ℃ after RNA extraction.
Grouping standard: a. primary gastric cancer is diagnosed before operation, and the cancer is not treated; b. no other malignancies were merged; c. no other complications such as gastric perforation, gastrointestinal hemorrhage, gastrointestinal obstruction and the like exist, and the general condition of the patient before the operation is good; d. no other chronic diseases, such as hypertension, diabetes, etc.
2. Detection at the transcriptional level
Total RNA was extracted from each of the experimental group (gastric cancer tissue) and the control group (adjacent normal stomach tissue) using RNAassoplus (TaKaRa, Shiga, Japan), and the concentration and purity of the RNA were measured using a spectrophotometer. It was reverse-transcribed into complementary DNA (cDNA) using PrimeScript reverse transcriptase (TaKaRa, Shiga, Japan). cDNA as template, according to SYBRGREEN EX TaqTM(TaKaRa, Shiga, Japan) Specification requires the reaction system and reaction time to carry out QPCR on LightCycler480, and all experimental results were repeated 3 times with results of 2－△△CT calculations and statistical analysis. GAPDH was used as the reference gene in the experiment. An upstream primer 5'-GGCAGTAGCATTATGAACAA-3' (SEQ ID NO.4) and a downstream primer 5'-ACAGAGCAAGACTCCATC-3' (SEQ ID NO.5) of LINC 01012; GAPDH upstream primer: 5'-TCATGGGTGTGAACCATGAGAA-3' (SEQ ID NO.6), and a downstream primer 5'-GGCATGGACTGTGGTCATGAG-3' (SEQ ID NO. 7).
3. Results
The statistical results show that the expression level of LINC01012 in gastric cancer tissues (relative expression level of 2.567 + -0.702) is significantly increased, about 2.6 times, compared with the paracarcinoma tissues (setting expression level of 1), and the difference has statistical significance (P < 0.05). Using the GEO database for electronic validation, ROC curve analysis showed that LINC01012 has an AUC value of 0.8319 when differentiating between gastric cancer patients and normals.
Example 2 inhibition of LINC01012 expression
1. Cell culture and transfection
Cell culture: the gastric cancer cell line BGC-823 is prepared from DMEM containing 10% FBS and placed in 5% CO2And saturated humidity, and culturing in a carbon dioxide incubator at 37 ℃. The culture medium was changed every two days and the cells were passaged by digestion with 0.25% trypsin.
siRNA transfection: the day before transfection, cells are digested and plated onto a petri dish or plate in an amount that ensures a density of 30-50% at the next day of transfection. The siRNA transfection was performed strictly according to LipofectaminTM2000, the culture was continued for 48h after 4-6h by replacing fresh culture medium containing 10% FBS.
2. SiRNA design
Design of siRNA (small interfering RNA): by BLAST search, siRNA sequences were designed in the specific sequence region of LINC 01012:
siRNA-LINC01012
sense strand: 5'-UUUCUUCUGAACAAGAAUGUG-3' (SEQ ID NO. 8);
antisense strand: 5'-CAUUCUUGUUCAGAAGAAACA-3' (SEQ ID NO. 9).
The above siRNA was synthesized by Shanghai Jima pharmaceutical technology, Inc., while the company provided negative control siRNA (siRNA-NC).
3. Detection of siRNA interference Using QPCR assay
The procedure is as in example 1.
4. Results
After siRNA transfects gastric cancer cell strain BGC823, change of LncRNA transcription level is detected by QPCR. The results are shown in fig. 1, the expression level of LncRNA in the experimental group cells (siRNA-LncRNA) is obviously reduced compared with the control group cells (siRNA-NC), the inhibition rate is about 73%, the difference has statistical significance (P <0.05), and the results show that the siRNA-LncRNA of the invention can obviously inhibit the expression of LINC01012 in the cells.
Example 3 measurement of gastric cancer cell proliferation potency by inhibiting LINC01012 expression
1. The method comprises the following steps:
BGC-823 cells in logarithmic growth phase are taken and inoculated on a 96-well culture plate, after the cells are cultured for 12 hours, siRNA-LncRNA and siRNA-NC are transfected respectively, and after 4-6 hours, DMEM containing 10% FBS is replaced to continue culturing for 72 hours; and adding 10 mu LCCK8 to each well of each plate, incubating for 2h in an incubator, and detecting the absorbance value with the wavelength of 450nm by using a microplate reader. Each group of cells was set with 5 replicate wells, and the experiment was repeated 3 times to plot cell growth curves.
2. Statistical analysis
The experimental data are expressed as mean-squared-off standard deviations (MS-SD) using SPSS15.0 statistical software, and subjected to one-way analysis of variance (ANOVA) or t-test. P <0.05 is statistically significant for the differences.
3. Results
The results showed that 72h after transfection, the cell growth rate of siRNA-LncRNA group (0.874 + -0.024) was significantly decreased (P <0.05) compared to siRNA-NC (1.546 + -0.052). The result shows that the inhibition of LINC01012 expression has an inhibitory effect on the proliferation capacity of the gastric cancer cell strain BGC 823.
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.
Sequence listing
<110> Beijing, the deep biometric information technology GmbH
Application of LINC01012 as gastric cancer diagnosis molecule and drug target gene
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<170>SIPOSequenceListing 1.0
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tggccccgga gcctacaacc tgacgcaggt ggacaaggtg gaggagagag aattccttgg 180
cagtagcatt atgaacaaat gcagggctga gaaacagcat ggtggccttg actaagttta 240
tggcgacagc agttggggta acatgtgacc tggatctgat cattgtgatt caagtcagcg 300
tcatttaggg ttttcgggaa acattttttg aaagggagca acttggctgg aacactccat 360
caattctttg ccctgtgcat ttttgccctt ctttctgcct ctcaggaatg cagagagaat 420
tcctggagga acagcagcta tcatacatcc gtgatgaaag ccatgggaca aaggcaattg 480
agcaagaaga caggagctgg ggacattctt gacagtagtg ccacctctgg atgatttacc 540
ttgggacctc ttattaaatg agataggact tcttcctgtt taatccactg gagtggaatt 600
ttctgttaca gtgaatgcag tcctaattga tgatattggc taatacatat tagcacctgc 660
tatgtcccag tacagtttaa gcattttatc tactaatgtt ttaaattttc tcagtgacct 720
tatgagatag gccaaaggtt ctcagagtgt ggttcctgga ccagcaacat cagcatcacc 780
tggaaacttg ttagaaatac aaattttagg gcctggccct agacccactg aatcagaaac 840
tctaggggtg aggacctagc agtctgttta aaaagccctc catgctattt tgatgaatgc 900
caaagttgga cagctactga gatagggtat tgttattatc cccattttac agataaggaa 960
acagctgaat ttatgtcact tgcccaatgt cacacaacta gcaagactga ctccttgaag 1020
ctcatcacat gtaagaactg ctttagagca gtggtcttca aactttgaaa gcatatcctg 1080
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aagtttaaac agttgtaaaa aatgtgattt ctggtatatt gtgaattatg acattttaaa 1200
atgtattcca tcactcctct aaatgtattc agtggtttct aaataccata gtgatttaaa 1260
tctatcatca atttaaaaaa aaaaacaagc tttttttttt aacattagat attttgcatc 1320
tttttctctc tttttttttt tttttttgag acagactctc actgtgtcac ccaggctgga 1380
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ctcaagtgat ctgcccacct tggtcttcca aagtactggg actacaggca tgagaaacca 1620
agcccggcca tctttctctt ttttacttga attcagtctt tttattcttc ttccattcct 1680
ctctctctct ctcttttttt ttttgagaca gtctcactat gtcacccagg ctggagtgca 1740
gtggcgcaat ctcggctcac tgcagcctct acctcccaga ttcaagcgat tctcctgcct 1800
cagcctcctg agtagttggg actacaggtg cgtgccacca tgcccggcta atttttttgt 1860
atttttagta gagatggggt ttcaccattt tggccaggat ggtctcaatc tcctgacctc 1920
gtgatctgcc tgccttggcc tcccaaagtg ctgggatcac aggcatgaac caccgctcct 1980
ggcctctttc tttcttttct gttcttttcc atctgccctc cccaaacatc tccctcccta 2040
cctccctccc tccctacttt ccttgcttcc ttccttcctc tctttctgtt tctctatttt 2100
tctttttttc cttctttttt gagaccatct cactctgttg cccaggatgg aatgcagtgg 2160
cacaatctta gctcactaca acctctgcct cccaggttca agcaattctc ctgcctcagc 2220
ctcccaagta gctgggacca caggcagatg ccaccacacc aagctaaagt ttttttttta 2280
tttttcatag agacagggga tctcattatg ttgcccaaac tggtcgtgaa ctcctgggct 2340
caagtgctct gcctgcctcg gcctcccaaa gtgtagggat tacaggcgtg agccactgtg 2400
cacgttggga ctaatatttt cttattcgta tttccatggc agaatattcc tcactgctag 2460
aatgagacca gaggtcaaaa tcagctctat gtctgtttgt cattcttatg gatatgagaa 2520
attgtgttca cacaagtaga taggaatgaa aggaattttg aaatagaaat gcagttctaa 2580
tttttctaac ttccatatta cgtgccaaaa atcacataat aatctatcat aagattctct 2640
agtcctctct catctggtga tcttcttttc cacgctttct ttgcaacccc ctcatgtggc 2700
caacatttag agtaatgttt tccaacttct ttttcttcag ctcacagtaa taaatacatt 2760
ttacatcttg aaaaaaaaaa aaaaa 2785
<210>2
<211>675
<212>DNA
<213> human source (Homo sapiens)
<400>2
cgccagggga tgcaaaccct tctgcccctt tttgctaccc ctgaagctta ctgttagtgg 60
tcgagttttt ctggaagtgg gcttccgcga tcgcggggac caacgccctg agctcttgtg 120
cccaaagggg atcgccaggt tttttgatgc ggatcgaaag ggactggccc cggagcctac 180
aacctgacgc aggtgggact tggggaaatg tcgagaaacc taggtctggg acttgagttg 240
gcttccaggg cacacattct tgttcagaag aaacacattg caaaaatcgg gaggtgacgg 300
gcagatgaag tgtcggcgct tcgcctcggg actccagtgc aacctatggg gcttttggaa 360
atgaggaagt cgatgactct gcagggactc ctctgaccca ggagcagatg gcgctcgtct 420
gggcactccc caggagaaaa ggtgcatcgg gaagcagcct ggcttgtgga tggtggacaa 480
ggtggaggag agagaattcc ttggcagtag cattatgaac aaatgcaggg ctgagaaaca 540
gcatggtggc cttgactaag tttatggcga cagcagatgg agtcttgctc tgtcgcccag 600
gctagagtgc agtggcgcga tctcagctca ctgaaacctc tgcctcccgg gttcaagtga 660
ttctcctgcc tccgc 675
<210>3
<211>753
<212>DNA
<213> human source (Homo sapiens)
<400>3
cgccagggga tgcaaaccct tctgcccctt tttgctaccc ctgaagctta ctgttagtgg 60
tcgagttttt ctggaagtgg gcttccgcga tcgcggggac caacgccctg agctcttgtg 120
cccaaagggg atcgccaggt tttttgatgc ggatcgaaag ggactggccc cggagcctac 180
aacctgacgc aggtgggact tggggaaatg tcgagaaacc taggtctggg acttgagttg 240
gcttccaggg cacacattct tgttcagaag aaacacattg caaaaatcgg gaggtgacgg 300
gcagatgaag tgtcggcgct tcgcctcggg actccagtgc aacctatggg gcttttggaa 360
atgaggaagt cgatgactct gcagggactc ctctgaccca ggagcagatg gcgctcgtct 420
gggcactccc caggagaaaa ggtgcatcgg gaagcagcct ggcttgtgga tggtggacaa 480
ggtggaggag agagaattcc ttggcagtag cattatgaac aaatgcaggg ctgagaaaca 540
gcatggtggc cttgactaag tttatggcga cagcagataa ggaaacagct gaatttatgt 600
cacttgccca atgtcacaca actagcaaga ctgactcctt gaagctcatc acatatggag 660
tcttgctctg tcgcccaggc tagagtgcag tggcgcgatc tcagctcact gaaacctctg 720
cctcccgggt tcaagtgatt ctcctgcctc cgc 753
<210>4
<211>20
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>4
ggcagtagca ttatgaacaa 20
<210>5
<211>18
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>5
acagagcaag actccatc 18
<210>6
<211>22
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>6
tcatgggtgt gaaccatgag aa 22
<210>7
<211>21
<212>DNA
<213> Artificial Sequence (Artificial Sequence)
<400>7
ggcatggact gtggtcatga g 21
<210>8
<211>21
<212>RNA
<213> Artificial Sequence (Artificial Sequence)
<400>8
uuucuucuga acaagaaugu g 21
<210>9
<211>21
<212>RNA
<213> Artificial Sequence (Artificial Sequence)
<400>9
cauucuuguu cagaagaaac a 21

Claims (10)

1. The application of the reagent for detecting the expression of the long-chain non-coding RNA in the preparation of gastric cancer diagnosis products; the long non-coding RNA is LINC 01012.
2. The use of claim 1, wherein the reagent comprises PCR amplification primers used for detecting the expression level of the long non-coding RNA by using SYBR Green, TaqMan probes, molecular beacons, two-hybrid probes, or composite probes.
3. The use according to claim 2, wherein the primer sequences are shown as SEQ ID No.4 and SEQ ID No. 5.
4. The use according to any one of claims 1 to 3, wherein the product comprises a kit, chip, or strip.
5. A product for gastric cancer diagnosis comprising an agent for detecting the expression of the long non-coding RNA of any one of claims 1 to 4.
6. The product of claim 5, wherein the reagent comprises SYBR Green, TaqMan probes, molecular beacons, two-hybrid probes, or PCR amplification primers used in the detection of the expression level of the long non-coding RNA.
7. The product of claim 6, wherein the primer sequences are shown as SEQ ID No.4 and SEQ ID No. 5.
8. A pharmaceutical composition for treating gastric cancer, comprising an agent that inhibits the long non-coding RNA of any one of claims 1-4.
9. The pharmaceutical composition of claim 8, wherein the agent comprises an agent that inhibits the level of long-chain non-coding RNA, or an agent that inhibits the functional activity of the long-chain non-coding RNA.
10. Use of the long non-coding RNA of any one of claims 1-4 in the manufacture of a medicament for the treatment of gastric cancer.
CN201911399582.4A 2019-12-30 2019-12-30 Application of LINC01012 as gastric cancer diagnosis molecule and drug target gene Withdrawn CN110951882A (en)

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Application Number Priority Date Filing Date Title
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