CN116904588A - Application of LINC01633 in diagnosis and treatment of gastric cancer - Google Patents

Application of LINC01633 in diagnosis and treatment of gastric cancer Download PDF

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CN116904588A
CN116904588A CN202210966673.7A CN202210966673A CN116904588A CN 116904588 A CN116904588 A CN 116904588A CN 202210966673 A CN202210966673 A CN 202210966673A CN 116904588 A CN116904588 A CN 116904588A
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linc01633
gastric cancer
expression
screened
gene
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沈贤
郑建建
陈孝冬
陈吉
詹雅婷
李鑫淼
林丽凡
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First Affiliated Hospital of Wenzhou Medical University
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Abstract

The invention discloses a biomarker for diagnosis and treatment of gastric cancer, and the specific biomarker is LINC01633. The invention discovers that the differential expression of LINC01633 is related to the occurrence and development of gastric cancer for the first time, and the expression of LINC01633 is up-regulated in gastric cancer patients, so that the intervention of LINC01633 gene expression can become a new way for treating gastric cancer. To further verify, it was found that by reducing the expression of LINC01633 by the inhibitor, proliferation of gastric cancer cells and the number of cell-forming clonal colonies can be significantly reduced.

Description

Application of LINC01633 in diagnosis and treatment of gastric cancer
Technical Field
The invention relates to the technical field of biological medicines, in particular to application of LINC01633 in diagnosis and treatment of gastric cancer.
Background
Gastric Cancer (GC) is one of the most common malignancies worldwide, with the incidence being the fifth most common among all malignancies worldwide, the third leading cause of death in cancer patients. Gastric cancer patients can be classified into three subtypes according to the Lauren system classification: high differentiation (non cardiac/intestinal), low differentiation (cardiac/diffuse), and mixed. Highly differentiated (intestinal) gastric cancer is most common, and is most common in men older than 70 years of age, with a large tumor volume. This subtype generally has a better prognosis than the poorly differentiated (diffuse) subtype. The survival rate of the poorly differentiated subtype is low, often in young women. Mixed subtypes are often present in a subset of older male patients, and are highly invasive and metastatic. In addition to Lauren's classification, the World Health Organization (WHO) classifies GC into four main types according to major histological patterns: tubular adenocarcinoma, papillary adenocarcinoma, mucous adenocarcinoma, and mucous adenocarcinoma. Currently, surgical treatment is the primary choice for clinically treating gastric cancer, but recurrence and metastasis are also common. For the treatment of gastric cancer, there are still many unknown studies and researches on the pathogenesis of gastric cancer are critical to reveal the nature of gastric cancer and develop new treatments.
LncRNA is defined as transcripts that are more than 200 nucleotides in length and cannot be translated into proteins. In various types of cancer cells, the abnormal expression of lncRNA plays a vital role. In general, lncRNA plays a regulatory role in gene expression at different levels, including chromatin modification, transcription, and post-transcription. Mechanically, lncRNA can exert its regulatory functions through interactions with DNA, mRNAs, ncRNAs and proteins, acting as different functional molecules in various cancer-associated cellular processes, serving as signals, baits, scaffolds, and guides, etc. IT has now been found that a variety of lncrnas are closely related to gastric cancer, such as lncRNA SNHG1, lncrrnasnd 1-IT1, and lncrrnadleu 2, among others.
At present, related regulation and control mechanisms of the lncRNA in the gastric cancer are still to be further elucidated, the elucidation of the gastric cancer mechanism, diagnosis, treatment, prevention and the like of the gastric cancer are all important by searching for the related lncRNA in the gastric cancer, and a new thought can be provided for gene targeting treatment of the gastric cancer.
Disclosure of Invention
In order to overcome the defects of the prior art, the invention aims to provide a biomarker related to generation and development of gastric cancer, and the early diagnosis of gastric cancer and the accurate molecular targeted treatment of gastric cancer are realized by detecting the biomarker.
In order to achieve the above purpose, the present invention adopts the following technical scheme:
the invention provides application of a reagent for detecting LINC01633 in preparation of a product for diagnosing gastric cancer.
The reagent comprises a reagent for detecting the expression level of LINC01633 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 LINC01633 by real-time quantitative PCR comprises at least a pair of primers for specifically amplifying LINC01633.
Further, the reagent comprises:
a probe that specifically recognizes LINC 01633; or (b)
A primer that specifically amplifies LINC01633.
Further, the primer sequences of the specific amplified LINC01633 are shown in SEQ ID NO.1 and SEQ ID NO. 2.
The invention provides a product for diagnosing gastric cancer, which comprises a reagent for detecting the expression level of LINC01633 in a sample.
The product comprises a chip, a formulation or a kit. Wherein the chip comprises a solid support and an oligonucleotide probe immobilized on the solid support, wherein the oligonucleotide probe comprises an oligonucleotide probe for LINC01633 for detecting the transcript level of LINC 01633; the kit comprises a primer or a chip for detecting the LINC01633 transcription level.
The solid support may be made of various materials commonly used in the field of gene chips, such as, but not limited to, nylon membranes, glass or silicon wafers modified with reactive groups (e.g., aldehyde groups, amino groups, etc.), unmodified glass slides, plastic sheets, etc.
The gene detection kit or the gene chip can be used for detecting the expression level of a plurality of genes (such as a plurality of genes related to gastric cancer) including the LINC01633 gene, and can be used for simultaneously detecting a plurality of markers of gastric cancer, so that the accuracy of gastric cancer diagnosis can be greatly improved.
Further, the reagent for detecting the level of LINC01633 in the sample comprises:
a probe that specifically recognizes LINC 01633; or (b)
A primer that specifically amplifies LINC01633.
Further, the primer sequences for specifically amplifying LINC01633 are shown as SEQ ID NO.1 and SEQ ID NO. 2.
The invention provides an application of LINC01633 in preparing a pharmaceutical composition for treating gastric cancer.
Further, the pharmaceutical composition comprises an inhibitor of LINC01633, and/or other drugs compatible with the inhibitor and pharmaceutically acceptable carriers and/or excipients, wherein the inhibitor can inhibit the expression of LINC01633 or substances related to the upstream or downstream pathway of LINC01633. The inhibitor may act at the DNA level or at the RNA (i.e., gene product) level.
Further, the inhibitor is an siRNA against LINC01633.
In the invention, when effective siRNA sequences are screened, the optimal effective fragments are found out through a large number of comparison and analysis. In the specific embodiment of the invention, the inventor designs and synthesizes a plurality of siRNA sequences, and respectively transfects gastric cancer cell lines through transfection reagents to verify, so that the interference molecules with better interference effects are detected, and respectively have sequences shown by SEQ ID NO.9 and SEQ ID NO.10, and further, the inhibition efficiency is very high for the cell experiments through the cell level experiments.
The nucleic acid inhibitors of the invention, such as siRNA, may be chemically synthesized or prepared by transcription of an expression cassette in a recombinant nucleic acid structure into single stranded RNA. Nucleic acid inhibitors such as siRNA can be delivered into cells by use of an appropriate transfection reagent, or can also be delivered into cells using a variety of techniques known in the art.
Those skilled in the art will recognize that the utility of the present invention is not limited to quantifying the 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 LINC01633 has a sequence as shown in the LINC01633 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, the probe is able to bind to a target polynucleotide that lacks complete sequence complementarity with the probe. Probes may be labeled directly or indirectly, and include primers. Hybridization means, 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, dextrose, urea, starch, water and the like, fillers such as starch, sucrose and the like; binders such as simple syrups, dextrose solutions, starch solutions, 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 promoters quaternary ammonium compounds, sodium lauryl sulfate, and the like; surfactants such as polyoxyethylene sorbitan fatty acid ester, sodium lauryl sulfate, monoglyceride of stearic acid, cetyl alcohol, etc.; wetting agents such as glycerin, starch, and the like; adsorption carriers such as starch, lactose, bentonite, silica gel, kaolin, and bentonite; 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, isotonic 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, bucally, vaginally, or by 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 excipients. In some cases, a pharmaceutically acceptable acid, base or buffer may be used to adjust the pH of the formulation to improve the stability of the formulated compound or dosage form thereof. The term parenteral as used herein includes subcutaneous, intradermal, intravenous, intramuscular, intraarticular, intraarterial, intrasynovial, intrasternal, intrajugular, intralesional, and intracranial injection or infusion techniques. The pharmaceutical composition of the present invention may be administered to the subject by any route so long as the target tissue is achieved.
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, common carriers include lactose and corn starch. A lubricant such as magnesium stearate is 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, some sweetener and/or flavoring and/or coloring agents may be added. Dosage unit formulations for oral administration may be microencapsulated, as appropriate. The formulations may also be prepared to have prolonged or sustained release, for example, by coating or embedding particulate matter in polymers, waxes, and the like. The pharmaceutical composition of the invention can be used for reducing the overexpression of endogenous LINC01633, and can be used for treating gastric cancer caused by up-regulation of LINC01633 by reducing the expression of LINC01633.
In the present invention, a compound that inhibits expression of LINC01633 can be administered to a subject as naked RNA together with a delivery agent as a nucleic acid (e.g., a recombinant plasmid or viral vector) comprising a sequence that inhibits expression of LINC01633. 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 anticancer agents. In a specific embodiment, the pharmaceutical composition comprises at least one compound that inhibits expression of the LINC01633 gene and at least one chemotherapeutic agent. Chemotherapeutic agents useful in the present invention include, but are not limited to: microtubule activators, alkylating agents, antineoplastic antimetabolites, platinum-based compounds, DNA-alkylating agents, antitumor antibiotics, antimetabolites, tubulin stabilizers, tubulin destabilizers, hormone antagonists, topoisomerase inhibitors, protein kinase inhibitors, HMG-COA inhibitors, CDK inhibitors, cyclin inhibitors, caspase inhibitors, metalloprotease inhibitors, antisense nucleic acids, triple helix DNA, nucleic acid aptamers, and molecularly modified viral, bacterial, and exotoxic agents.
The medicaments of the present invention may also be used in combination with other medicaments for the treatment of gastric cancer, and other therapeutic compounds may be administered simultaneously with the main active ingredient, even in the same composition. Other therapeutic compounds may also be administered alone in separate compositions or in dosages different from the primary active ingredient. A partial dose of the principal component 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 may be adjusted during the course of treatment according to the severity of the symptoms, the frequency of recurrence and the physiological response of the treatment regimen.
The pharmaceutical composition of the present invention may be formulated into ointments, creams, suspensions, lotions, powders, solutions, pastes, gels, sprays, aerosols or oils for topical administration.
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 in a reasonable benefit/risk ratio applicable to medical treatment or prevention, and the effective amount level may be determined according to factors including the severity of the disease, the activity of the drug, the age, weight, health, sex, sensitivity of the patient to the drug, the administration time, administration route and rate of discharge, the treatment time of the composition of the present invention used, factors of the drug used in combination with or simultaneously with the composition of the present invention, and other factors known in the medical arts. The pharmaceutical composition of the present invention may be administered as a single therapeutic agent or concurrently with other therapeutic agents, and may be administered sequentially or simultaneously with conventional therapeutic agents. Furthermore, the administration may be performed in a single or multiple times. It is important that the above-mentioned factors be considered and that the administration be carried out in such an amount that the maximum effect can be obtained with the minimum amount without side effects.
The term "treatment" refers to not the purpose of cure, but rather to slowing (reducing) the targeted pathological condition or disorder or preventing recurrence. If the patient is successfully "treated" after receiving a therapeutically effective amount of the therapeutic agent, the patient exhibits a observable and/or measurable reduction or disappearance of signs and symptoms of one or more specific diseases. For example, a significant reduction in the number of cancer cells or the disappearance of cancer cells, a reduction in tumor size; inhibit (i.e., slow down, and preferably stop to some extent) tumor metastasis; inhibit tumor growth to some extent; increasing the time to reduce and/or alleviate to some extent one or more symptoms associated with a particular cancer; reduced morbidity and mortality, and improved quality of life. The alleviation of signs or symptoms of a disease can be perceived by a 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 to be treated if they feel stable.
In the present invention, the term "sample" includes, but is not limited to, cells, tissues, organs, body fluids (blood, lymph, etc.), digestive fluids, expectoration, lung cell bronchial washes, urine, feces, etc. Preferably, the sample is tissue or blood.
The invention also provides application of LINC01633 in screening substances to be screened for preventing or treating gastric cancer.
Further, the step of screening the substance to be screened for preventing or treating gastric cancer comprises:
the substances to be screened are treated to express or contain a system of LINC01633 genes; and
detecting expression of the LINC01633 gene in said system;
wherein, if the substance to be screened can reduce the expression or activity of the LINC01633 gene (preferably, the expression or activity is significantly reduced, such as by more than 20%, preferably by more than 50%, more preferably by more than 80%), the substance to be screened is a substance to be screened for preventing or treating gastric cancer.
Further, the systems described above include (but are not limited to): a cellular 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 gastric cancer, so that patients can be treated in early stage, and further survival rate and life treatment are improved. The invention also provides a treatment means and a pharmaceutical composition for realizing accurate molecular treatment of gastric cancer. The invention provides a method for screening substances to be screened for preventing or treating gastric cancer, which has important roles in diagnosis, prognosis and prediction of gastric cancer.
Drawings
FIG. 1 is a diagram showing the detection of LINC01633 expression in gastric cancer patients by QPCR;
FIG. 2 is a diagram showing the detection of LINC01633 expression in gastric cancer cells by QPCR;
FIG. 3 is a graph showing the effect of QPCR detection of transfected siRNA on LINC01633 expression in KATO III gastric cancer cells;
FIG. 4 is a graph showing the effect of LINC01633 on KATO III gastric cancer cell proliferation using CCK 8.
Detailed Description
The invention will now be described in further detail with reference to the drawings and examples. The following examples are only illustrative of the present invention and are not intended to limit the scope of the invention. The experimental procedure, without specific conditions noted in the examples, is generally followed by conventional conditions, such as Sambrook et al, molecular cloning: conditions described in the laboratory Manual (New York: coldSpringHarborLaboratory Press, 1989) or as recommended by the manufacturer.
Example 1: screening for Gene markers associated with gastric cancer
1. Sample collection
Each of 50 cases of gastric cancer patients was collected for their cancerous tissue and paracancerous tissue, informed consent was obtained from the patients, and all of the specimens were obtained by consent from the tissue ethics committee.
2. Preparation of RNA samples
Tissue RNA was extracted using QIAGEN tissue RNA extraction kit, and the procedure was performed according to the specific procedures described.
3. Mass analysis of RNA samples
The concentration and purity of the extracted RNA were measured using Nanodrop2000, the integrity of the RNA was measured by agarose gel electrophoresis, and the RIN value was measured by Agilent 2100. The concentration is more than or equal to 200 ng/. Mu.l, and the OD260/280 is between 1.8 and 2.2.
4. rRNA removal
Ribosomal RNA was removed from the total RNA using the Ribo-Zero kit.
5. Construction of cDNA library
Construction of the cDNA library was performed using TruseqRNAsamplePrepKit from Illumina, and the specific procedure was according to the instructions.
6. Sequencing on machine
The cDNA library was sequenced using a Hiseq4000 sequencing platform, the specific procedure being according to the instructions.
7. High throughput transcriptome sequencing data analysis
Performing bioinformatics analysis on the original count data by adopting R package DESeq2, deleting lncRNA which is not easy to detect, and differentially expressing lncRNA screening standards: FDR <0.05, abs (log 2 FC) >2.
7. Results
The RNA-seq results showed that LINC01633 expression levels in gastric cancer tissues were significantly higher than in paracancerous tissues.
Example 2: QPCR sequencing verified differential expression of LINC01633 gene.
1. Large sample QPCR validation was performed on LINC01633 gene differential expression. 50 samples of stomach cancer tissue and other tissue were collected in the same manner as the sample collection in example 1.
2. The RNA extraction procedure was as in example 1.
3. Reverse transcription:
using 20 μl reaction system, 1 μg total RNA per sample was used as template RNA, and the following components were added to the PCR tube: DEPC water, premix reagent, template RNA. Maintained at 37℃for 15min,50℃for 5min,98℃for 5min,4 ℃.
(3) qPCR amplification assay
Primers were designed based on the sequences of the LINC01633 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 LINC01633 gene is as follows:
forward primer: 5'-GCAAAATGTCCGCATCTCCC-3' (SEQ ID NO. 1) reverse primer: 5'-GGAGGAGGCATGTCAGCTTT-3' (SEQ ID NO. 2)
The primer sequences for housekeeping gene GAPDH are:
forward primer: 5'-GAAAGCCTGCCGGTGACTAA-3' (SEQ ID NO. 3) reverse primer: 5'-GCCCAATACGACCAAATCAGAG-3' (SEQ ID NO. 4)
The following reaction system was prepared in 10. Mu.l: SYBRGreen polymerase chain reaction system 5. Mu.l, forward and reverse primer (5. Mu.M) 2. Mu.l each, template cDNA 1. Mu.l. Each operation was performed on ice. Each sample is provided with 3 parallel tubes, and all amplification reactions are repeated more than three times to ensure the reliability of the results. The amplification procedure was: 95℃for 60s, (95℃for 10s,60℃for 30 s). Times.40 cycles.
SYBRGreen 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 by a delta CT method.
3. Statistical method
Experiments were performed in 3 replicates, and the data obtained were expressed as mean ± standard deviation, and were statistically analyzed using SPSS18.0 statistical software, the differences between the two using t-test, and were considered statistically significant when P < 0.05.
4. Results
As shown in FIG. 1, the expression of LINC01633 gene was up-regulated in gastric cancer tissues compared to paracancerous tissues, and the difference was statistically significant (P < 0.05) consistent with RNA-sep results.
Example 3: differential expression of LINC01633 gene in gastric cancer cell line.
1. Cell culture
Human gastric cancer cell lines AGS, KATOIII and normal gastric epithelial cell line GES-1 in culture medium DMEM or 1640 containing 10% fetal bovine serum and 1% P/S at 37deg.C and 5% CO 2 Culturing in an incubator with a relative humidity of 90%. The solution was changed 1 time from 2 to 3 days and passaged by routine digestion with 0.25% trypsin containing EDTA.
2. Extraction of RNA
RNA in cells was extracted using Qiagen's cellular RNA extraction kit, and experimental procedures were performed according to the instructions.
3. The reverse transcription was performed in the same manner as in example 2
4. Statistical method
Experiments were performed in 3 replicates, and the data obtained were expressed as mean ± standard deviation, and were statistically analyzed using SPSS18.0 statistical software, the differences between the two using t-test, and were considered statistically significant when P < 0.05.
5. Results
As shown in FIG. 2, the expression of LINC01633 gene was up-regulated in both gastric cancer cells AGS and KATOIII, and the difference was statistically significant (P < 0.05) as compared with the normal gastric epithelial cell line, consistent with the RNA-sep results.
Example 4: silencing of the LINC01633 Gene
1. Cell culture
The human gastric cancer cell line KATOIII was cultured, and the procedure of example 3 was followed.
2. SiRNA design
Interference RNAs were designed according to the sequence of LINC01633, dividing the experiment into four groups: blank (KATOIII), negative control (siRNA-NC) and experimental (siRNA 1, siRNA2, siRNA 3), wherein the negative control siRNA has no sequence homology to LINC01633 gene. The designed siRNA sequence is shown below:
negative control siRNA sequence (siRNA-NC):
the sense strand is 5'-ACAAUAUCCACUUUACCAGAG-3' (SEQ ID NO. 5),
the antisense strand is 5'-CUGGUAAAGUGGAUAUUGUUG-3' (SEQ ID NO. 6);
siRNA1：
the sense strand is 5'-UUUCAUUUUUACUUGAAACUC-3' (SEQ ID NO. 7),
the antisense strand is 5'-GUUUCAAGUAAAAAUGAAAAU-3' (SEQ ID NO. 8);
siRNA2：
the sense strand is 5'-ACAAAUUCUGAAAUCUUAGUG-3' (SEQ ID NO. 9),
the antisense strand is 5'-CUAAGAUUUCAGAAUUUGUCU-3' (SEQ ID NO. 10);
siRNA3：
the sense strand is 5'-AUGGUUUAAAUAAACAAUGAA-3' (SEQ ID NO. 11),
the antisense strand was 5'-CAUUGUUUAUUUAAACCAUAG-3' (SEQ ID NO. 12), and the cells were grown at 2X 10 5 Plating into six-well cell culture plate at 37deg.C with 5% CO 2 Culturing cells in an incubator for 24 hours; transfection was performed in DMEM medium without diabody, containing 10% fbs, according to the instructions of liposome transfection reagent 2000 (purchased from Invitrogen).
3. QPCR (quantitative polymerase chain reaction) detection of transcript level of LINC01633 gene
3.1, extraction of total RNA from cells the specific procedure is the same as in example 3.
3.2, reverse transcription procedure was as in example 2.
3.3, QPCR amplification procedure was as in example 2.
4. Statistical method
Experiments were performed in 3 replicates, and the data obtained were expressed as mean ± standard deviation, and were statistically analyzed using SPSS18.0 statistical software, and differences between the interfering LINC01633 gene expression set and the control set were t-tested, and were considered statistically significant when P < 0.05.
5. Results
The results are shown in figure 3, in which LINC01633mRNA levels were reduced in the experimental group compared to transfected empty siRNA-NC, with the interference effect of siRNA2 group most pronounced, the differences being statistically significant (P < 0.05).
Example 5: CCK8 assay for cell proliferation
1. Cell culture and transfection procedure was as in example 4
2. CCK8 detection of cell proliferation
1) KATOIII cells from log proliferation phase were seeded in 96-well plates at 2X 10 per well 3 A cell; the experiments were divided into groups (control, siRNA-NC, siRNA 2) with 6 duplicate wells each;
2) 10 μl/well CCK8 reagent was added after 0h, 24h, 48h, 72h of transfection, respectively;
3) After 2h, the absorbance of A450 was measured using a microplate reader.
3. Statistical method
Experiments were performed in 3 replicates, using SPSS13.0 statistical software for statistical analysis, and the difference between the two was tested using t-test, which was considered statistically significant when P < 0.05.
4. Results
The results shown in fig. 4 show that: the cell growth rate of siRNA2 group is obviously lower than that of control group, the difference has statistical significance (P < 0.05), and the result shows that LINC01633 knockout can inhibit the growth of gastric cancer cells.
The above description of the embodiments is only for the understanding of the method of the present invention and its core ideas. It should be noted that it will be apparent to those skilled in the art that several improvements and modifications can be made to the present invention without departing from the principle of the invention, and these improvements and modifications will fall within the scope of the claims of the invention.

Claims (10)

1. Application of a reagent for detecting LINC01633 in preparing a product for diagnosing gastric cancer.
2. The use according to claim 1, characterized in that: the reagent comprises:
a probe that specifically recognizes LINC 01633; or (b)
A primer that specifically amplifies LINC01633.
3. The use according to claim 2, characterized in that: the primer sequences of the specific amplified LINC01633 are shown as SEQ ID NO.1 and SEQ ID NO. 2.
4. A product for diagnosing gastric cancer, characterized in that: the product includes reagents for detecting the expression level of LINC01633 in a sample.
5. The product according to claim 4, wherein: the product comprises a chip, a formulation or a kit.
Application of LINC01633 in preparing pharmaceutical composition for treating gastric cancer.
7. The use according to claim 6, characterized in that: the pharmaceutical composition comprises an inhibitor of LINC01633.
8. The use according to claim 7, characterized in that: the inhibitor is an siRNA against LINC01633.
Application of LINC01633 in screening substances to be screened for preventing or treating gastric cancer.
10. The use according to claim 9, characterized in that: the step of screening a substance to be screened for the prevention or treatment of gastric cancer comprises:
the substances to be screened are treated to express or contain a system of LINC01633 genes; and
detecting expression of the LINC01633 gene in said system;
wherein, if the substances to be screened can reduce the expression or activity of LINC01633 gene, the substances to be screened are proved to be substances to be screened for preventing or treating gastric cancer.
CN202210966673.7A 2022-08-12 2022-08-12 Application of LINC01633 in diagnosis and treatment of gastric cancer Pending CN116904588A (en)

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