CN110628915A - Application of lncRNA LSAMP-AS1 AS gastric cancer diagnosis marker - Google Patents
Application of lncRNA LSAMP-AS1 AS gastric cancer diagnosis marker Download PDFInfo
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- CN110628915A CN110628915A CN201911077377.6A CN201911077377A CN110628915A CN 110628915 A CN110628915 A CN 110628915A CN 201911077377 A CN201911077377 A CN 201911077377A CN 110628915 A CN110628915 A CN 110628915A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- 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
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
- C12Q2600/136—Screening for pharmacological compounds
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- 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
Abstract
The invention discloses application of lncRNA LSAMP-AS1 AS a gastric cancer diagnosis marker, relates to the technical field of medical biological detection, and provides application of lncRNA LSAMP-AS1 AS a gastric cancer diagnosis marker. The kit has good clinical application value and good detection effect on gastric cancer diagnosis.
Description
Technical Field
The invention belongs to the technical field of medical biological detection, and particularly relates to application of lncRNA LSAMP-AS1 AS a gastric cancer diagnosis marker.
Background
China is one of the countries with the highest incidence rate of gastric cancer, new cases of gastric cancer account for more than 40 percent of the world, and gastric cancer is the third cause of mortality of malignant tumors. In recent years, with the increasing of diagnosis and treatment technologies, the mortality rate of gastric cancer is reduced, the prognosis is obviously improved, and the 5-year survival rate of gastric cancer has been remarkably improved in the large medical center of China. There is still a considerable gap with respect to the data in japan. How to effectively improve the diagnosis and treatment level of gastric cancer is always a hot and difficult problem concerned by clinicians. Despite the increasing advances in medical technology, the survival rate of gastric cancer patients remains low, mainly because most patients have been diagnosed in the middle and advanced stages.
Expression profile gene chips are a common technique that has been recently developed for studying the expression profile of target genes between specimens, and a relatively complete system has been formed in the customization of chips, data acquisition and result analysis. Therefore, the gene chip plays an important role in researching the relationship between the mechanism of disease occurrence and development and gene change, and provides an effective method for researching the gene in the aspects of disease pathogenic process, diagnosis, treatment and the like. Currently, many groups have used this technique to study lncRNA associated with various tumors and screen lncRNA for correlation with tumor progression and prognosis.
lncRNA is a newly discovered class of non-coding RNA, which is over 200bp long and lacks an obvious Open Reading Frame (ORF), and can be classified into 5 classes according to the position of its chromosome, namely: sense lncRNA, antisense lncRNA, intergenic lncRNA, bidirectional lncRNA, and intragenic lncRNA. At present, there is a lot of evidence that lncRNA plays an important regulatory role in the biological properties of tumors through different mechanisms from transcription level to post-transcription level, such as interfering with transcription, activation or inactivation of proteins, transport or activation of transcription factors, modification of chromosomes, epigenetic regulation, etc.
Recently, the research on the lncRNA related to the gastric cancer has achieved remarkable results, and the research is now a hot spot of the research at home and abroad. Such as: lncrRNA GHET1 enhances the stability of c-Myc mRNA by the synergistic effect with insulin-like growth factor 2mRNA binding protein 1 (IGF 2BP1), thereby increasing the expression level and promoting the proliferation of gastric cancer cells. The lncRNA TINCR acts on KLF2 by combining with STAU1(staufen1) protein to protect mRNA thereof from degradation, and the increased expression of KLF2 directly acts on dependent cyclin kinase gene A (cyclin-dependent kinase gene A, CDKN1A/P21) and dependent cyclin kinase gene B (cyclin-dependent kinase gene B, CDKN2B/P15), thereby regulating proliferation and apoptosis of gastric cancer cells. In addition, lncrnaapvt 1 regulates p15 and p16 by synergistic action with EZH2, thereby promoting proliferation of gastric cancer cells. However, a number of abnormally regulated signaling pathways for gastric cancer-associated lncrnas are not yet known.
Disclosure of Invention
The invention aims to: the application of lncRNA LSAMP-AS1 AS a gastric cancer diagnosis marker is provided, and particularly the application of lncRNA LSAMP-AS1 AS a gastric cancer diagnosis marker and the application of a reagent for detecting the expression level of lncRNA LSAMP-AS1 in preparing a reagent or a kit for gastric cancer diagnosis are provided.
The technical scheme adopted by the invention is as follows:
use of lncRNA LSAMP-AS1 AS a diagnostic marker for gastric cancer.
The reagent for detecting the lncRNA LSAMP-AS1 is applied to the preparation of a detection reagent or a kit for gastric cancer diagnosis.
Further, the detection reagent or the kit is used for detecting the expression level of the lncRNA LSAMP-AS1 in the biological sample.
Further, the reagent or kit comprises: probes, gene chips, or PCR primers with detection specificity for lncRNA LSAMP-AS 1.
Further, PCR primers with detection specificity for lncRNA LSAMP-AS1 are AS follows:
an upstream primer: 5'-AAGGATGAGAAGAGCCACCA-3', respectively;
a downstream primer: 5'-GCACCAGGGTTGGATTCTT-3' are provided.
The lncRNA LSAMP-AS1 is applied to screening human gastric cancer diagnosis and treatment medicines.
Application of lncRNA LSAMP-AS1 in preparing a pharmaceutical composition for preventing or treating gastric cancer.
Further, the pharmaceutical composition comprises an enhancer of lncRNA LSAMP-AS1 and/or its expression products.
Many serum markers such as CEA, CA19-9, CA72-4, CA125, cancer antigen 242(CA242), and CA50 are widely used in the detection of gastric cancer. However, low sensitivity or low specificity reduces its clinical diagnostic value for gastric cancer.
Compared with invasive examination means such as gastroscope, the method for detecting the circulating lncRNA only needs to collect body fluid, has the characteristic of non-invasive detection, and greatly reduces the pain of a patient and the complexity of detection. Lncrnas have higher sensitivity and specificity, especially plasma exosome-type lncrnas, compared to conventional tumor markers such as CA19-9, CA125, CA72-4, CA242, CA50, CEA, and pepsinogen. Therefore, lncRNA can be used as a marker for early diagnosis of gastric cancer.
In conclusion, the lncRNA LSAMP-AS1 provided by the invention is used AS a gastric cancer diagnosis marker, has good specificity, and can accurately identify gastric cancer patients; the kit has high sensitivity, can detect cancer patients at an early stage, has high diagnosis efficiency and guaranteed reliability, provides a new clinical means for diagnosing the gastric cancer, has a simple and easy operation detection method, and has good transformation medical prospect.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings needed to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
FIG. 1 is a graph of the relative expression levels of lncRNA LSAMP-AS1 in gastric cancer tissues and their corresponding paraneoplastic tissues;
FIG. 2 is a graph showing the relative expression levels of lncRNA LSAMP-AS1 in gastric cancer cell lines.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the detailed description and specific examples, while indicating the preferred embodiment of the invention, are intended for purposes of illustration only and are not intended to limit the scope of the invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be derived by a person skilled in the art from the embodiments of the present invention without making any creative effort, shall fall within the protection scope of the present invention.
It is noted that relational terms such as "first" and "second," and the like, may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other identical elements in a process, method, article, or apparatus that comprises the element.
Use of lncRNA LSAMP-AS1 AS a diagnostic marker for gastric cancer.
The reagent for detecting the lncRNA LSAMP-AS1 is applied to the preparation of a detection reagent or a kit for gastric cancer diagnosis.
The detection reagent or the kit is used for detecting the expression quantity of lncRNA LSAMP-AS1 in the biological sample; the expression level of lncRNA LSAMP-AS1 was detected by real-time quantitative PCR technique.
The biological sample is selected from fresh tissue or cells of a subject, formalin-fixed or paraffin-embedded tissue or cells, blood or body fluid, and the like.
The reagent or kit comprises: probes, gene chips, or PCR primers with detection specificity for lncRNA LSAMP-AS 1.
The PCR primers with detection specificity for lncRNA LSAMP-AS1 were AS follows:
an upstream primer: 5'-AAGGATGAGAAGAGCCACCA-3', respectively;
a downstream primer: 5'-GCACCAGGGTTGGATTCTT-3' are provided.
The lncRNA LSAMP-AS1 is applied to screening human gastric cancer diagnosis and treatment medicines.
Application of lncRNA LSAMP-AS1 in preparing a pharmaceutical composition for preventing or treating gastric cancer.
The pharmaceutical composition comprises lncRNA LSAMP-AS1 and/or promoter of its expression product
The invention relates to a detection method for detecting lncRNA LSAMP-AS1 in serum, which comprises the following steps:
(1) tissue specimen Collection
Stomach cancer tissue: gastric cancer tissues with a diameter of about 0.5-1.0cm are cut from the tumor.
Tissue adjacent to the cancer: the distance between the tumor edge and the normal stomach tissue is more than 5cm, and the normal stomach tissue with the size of 0.5-1.0cm is cut.
In order to prevent RNA degradation, the specimen is cut immediately when the specimen is cut by an operation, washed twice by PBS, put into a freezing tube and immediately put into liquid nitrogen, and finally transferred to a refrigerator at minus 80 ℃ for long-term storage;
(2) extraction of Total RNA
1) Tissue homogenate
Taking out a sample to be detected, unfreezing the sample on ice, weighing 5g of gastric cancer tissue sample, mincing the gastric cancer tissue sample, adding 1mL of Trizol, and homogenizing the gastric cancer tissue sample for 2min in an ice tissue homogenizer at 8000r/min to fully grind tissues;
2) standing on ice for 10-15min to fully lyse cells;
3) adding 200 mul chloroform according to the proportion of 5:1, fully and uniformly mixing, and standing for 15min at room temperature;
4) centrifuging at 4 deg.C and 12000rpm for 15min until liquid demixing;
5) pipetting about 700. mu.l of the supernatant into a new EP tube;
6) adding 700 μ l isopropanol, mixing well, standing at room temperature for 30 min;
7) centrifuging at 4 deg.C and 12000rpm for 10min until precipitate appears at the bottom of the tube, and removing supernatant;
8) adding 1ml 75% ethanol, mixing, and standing at room temperature for 5 min;
9) centrifuging at 4 deg.C and 8000rpm for 10min, and removing supernatant;
10) opening the EP tube cover, and standing at room temperature for 10min to fully volatilize ethanol;
11) adding 50-100 μ l of 0.01% DEPC water, and gently blowing to dissolve RNA in DEPC water;
12) detecting the concentration and purity of the RNA by using a spectrophotometer;
(3) total RNA reverse transcription
1) The reaction system is as follows:
total RNA amount (0.5-5ng) X;
Oligo(dT)151 mu L of primer;
1 mu L of Random primer;
5×Buffer 5μL；
25mM MgCl2 4μL；
reverse transcriptase 1 μ L;
1 μ L of reverse transcriptase inhibitor;
PCR Nucleotide Mix 1.25μL；
3.75 mu L of non-nucleic acid water;
Total 25μL。
2) the reaction conditions were as follows:
72℃ 5min；
42℃ 60min；
72℃ 15min；
4℃ ∞。
the features and properties of the present invention are described in further detail below with reference to examples.
Example 1
The relative expression level of lncRNA LSAMP-AS1 in the gastric cancer tissue and the corresponding paracarcinoma tissue is tested, and the results are shown in fig. 1, which shows that the relative expression level of lncRNA LSAMP-AS1 in the gastric cancer tissue is significantly lower than that in the corresponding paracarcinoma tissue.
Example 2
The relative expression levels of lncRNA LSAMP-AS1 and lncRNA LSAMP in 5 gastric cancer cell lines SNU-1, BGC-823, MGC-803, AGS, SGC-7901, and normal gastric mucosal cell GES-1 were tested, respectively, and the results are shown in FIG. 2.
As can be seen from the figure, the relative expression level of lncRNA LSAMSP-AS 1 in the gastric cancer cell lines SNU-1 and MGC-803 is lower than that in normal gastric mucosal cells GES-1; the relative expression level of lncRNA LSAMP-AS1 in the gastric cancer cell lines BGC-823, AGS and SGC-7901 is higher than that in normal gastric mucosal cells GES-1.
The relative expression level of lncRNA LSAMP in the gastric cancer cell line SNU-1 is lower than that of normal gastric mucosal cell GES-1; the relative expression level of lncRNA LSAMP in the gastric cancer cell lines BGC-823, MGC-803, AGS and SGC-7901 is higher than that of normal gastric mucosal cells GES-1.
Example 3
The expression level of LSAMP-AS1 of 60 gastric cancer patients was tested, and the relationship between the expression level of LSAMP-AS1 and the clinical data of gastric cancer patients is shown in Table 1 below.
TABLE 1 relationship between LSAMP-AS1 expression level and clinical pathological characteristics of gastric cancer patients
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Sequence listing
Application of <120> lncRNA LSAMP-AS1 AS gastric cancer diagnosis marker
<160> 3
<170> SIPOSequenceListing 1.0
<210> 1
<211> 20
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 1
aaggatgaga agagccacca 20
<210> 2
<211> 19
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 2
gcaccagggt tggattctt 19
<210> 3
<211> 561
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 3
gaacagctcc ggtctacagc tcccagcgtg agcgacgcag aagacggtga tttctgcatt 60
tccatctgag gtggaatgac tgtggctctg tgccatctcg aaatctactg gtctctaagc 120
acaaacgaac attctcttaa actaatgaaa tgagtggcag gcagcgtgtg cagagcacca 180
gcaaaggctc agagggatgc ccacagagtt ccaccgcaac cgtgcacaga ggaggaggag 240
aatgctccct acaggtgaaa actaggctaa tacacaatga ttgaagacta aaggatgaga 300
agagccacca gtggggcgca ttcacagaga aacggtcaca tgatgaggca gcaagaaggc 360
aaaaattgcg agccaacaag agaggcctca gaagaatcca accctggtgc cgccttcatt 420
ttaaacttcc aacaacccca gaactgtgga aaaaaattct attttttgtc tgtggtattt 480
tgttatggca accctagcaa actaatacac tgagcaaact aatacattaa acagaatatt 540
aaaacgaaaa aaaaaaaaaa a 561
Claims (8)
- Use of lncRNA LSAMP-AS1 AS a diagnostic marker for gastric cancer.
- 2. Use of a reagent for detecting lncRNA LSAMP-AS1 according to claim 1 in the preparation of a gastric cancer diagnostic detection reagent or kit.
- 3. Use according to claim 2, characterized in that: the detection reagent or the kit is used for detecting the expression level of lncRNA LSAMMP-AS 1 in the biological sample.
- 4. The use according to claim 3, wherein said reagent or kit comprises: probes, gene chips, or PCR primers with detection specificity for lncRNA LSAMP-AS 1.
- 5. Use according to claim 4, characterized in that: the PCR primers with detection specificity to lncRNA LSAMMP-AS 1 are AS follows:an upstream primer: 5'-AAGGATGAGAAGAGCCACCA-3', respectively;a downstream primer: 5'-GCACCAGGGTTGGATTCTT-3' are provided.
- 6, the application of lncRNA LSAMP-AS1 in screening human gastric cancer diagnosis and treatment medicines.
- The application of lncRNA LSAMP-AS1 in preparing a pharmaceutical composition for preventing or treating gastric cancer.
- 8. Use according to claim 7, characterized in that: the pharmaceutical composition comprises an enhancer of lncRNA LSAMP-AS1 and/or its expression products.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108034722A (en) * | 2017-12-13 | 2018-05-15 | 广州医科大学附属肿瘤医院 | A kind of lncRNA SGOL1-AS1 are in the application as diagnosing gastric cancer marker |
| US20190161730A1 (en) * | 2016-07-07 | 2019-05-30 | Rubius Therapeutics, Inc. | Compositions and methods related to therapeutic cell systems expressing exogenous rna |
-
2019
- 2019-11-06 CN CN201911077377.6A patent/CN110628915A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20190161730A1 (en) * | 2016-07-07 | 2019-05-30 | Rubius Therapeutics, Inc. | Compositions and methods related to therapeutic cell systems expressing exogenous rna |
| CN108034722A (en) * | 2017-12-13 | 2018-05-15 | 广州医科大学附属肿瘤医院 | A kind of lncRNA SGOL1-AS1 are in the application as diagnosing gastric cancer marker |
Non-Patent Citations (3)
| Title |
|---|
| PENG QI ET AL.: "Reciprocal repression between TUSC7 and miR-23b in gastric cancer", 《INTERNATIONAL JOURNAL OF CANCER》 * |
| YUEHUI LIU ET AL.: "Construction and integrated analysis of crosstalking ceRNAs networks in laryngeal squamous cell carcinoma", 《BIOINFORMATICS AND GENOMICS》 * |
| 丽敏等: "胃癌分子标志物在临床中的研究进展", 《中国医学工程》 * |
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