CN113151456B - Diagnostic marker for head and neck cell carcinoma and application thereof - Google Patents
Diagnostic marker for head and neck cell carcinoma and application thereof Download PDFInfo
- Publication number
- CN113151456B CN113151456B CN202010075552.4A CN202010075552A CN113151456B CN 113151456 B CN113151456 B CN 113151456B CN 202010075552 A CN202010075552 A CN 202010075552A CN 113151456 B CN113151456 B CN 113151456B
- Authority
- CN
- China
- Prior art keywords
- lncrna
- head
- cell carcinoma
- seq
- squamous cell
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/7105—Natural ribonucleic acids, i.e. containing only riboses attached to adenine, guanine, cytosine or uracil and having 3'-5' phosphodiester links
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
-
- 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
-
- 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
- C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA
Abstract
The invention discloses a diagnostic marker for head and neck cell carcinoma, which is LncRNA-RP11-215P8.4, and has a nucleotide sequence shown as SEQ ID NO. 1 and a length of 669 bp. Also relates to the use of LncRNA-RP11-215P8.4 for the preparation of a kit for the diagnostic and/or prognostic evaluation of head and neck squamous cell carcinoma; also relates to an agent for preventing and/or treating head and neck squamous cell carcinoma, comprising an effective active ingredient in an amount that inhibits the expression of LncRNA-RP11-215P8.4 in cancer cells. For a patient suspected of suffering from head and neck squamous cell carcinoma, the detection reagent provided by the invention can be used for detection so as to facilitate early discovery and early treatment.
Description
Technical Field
The invention belongs to the technical field of biological detection, and particularly relates to a diagnostic marker for head and neck cell carcinoma and application thereof.
Background
Squamous Cell Carcinoma of Head and Neck (SCCHN) refers to squamous cell carcinoma that occurs in the oral cavity, pharynx, larynx, nasal cavity, sinuses, salivary glands, cervical airways and esophagus, and is one of the most common types of head and neck malignancies, which have high malignancy and poor prognosis. It is one of the leading causes of morbidity and mortality worldwide, accounting for approximately four percent of all cancers. In 2018, it is expected that about 51540 new patients with oral and throat cancer will die of 10030 patients with the disease. Alcohol and tobacco abuse are common causative factors in oral, oropharyngeal, hypopharyngeal, and laryngeal cancers, with a 5-year survival rate of only 63% in SCCHN patients, primarily because about 80-90% of advanced SCCHN patients develop local recurrence or distant metastasis. Therefore, the identification of novel biomarkers for early diagnosis of primary head and neck squamous cell carcinoma plays an important role in improving outcome.
LncRNA is an RNA that is more than 200 nucleotides in length and has no protein coding function. Studies have shown that high throughput chip and RNA sequencing technologies have identified large numbers of lncrnas that are aberrantly expressed in various cancers. With the advent of a continuously updated big data era, there has been a great progress in the research on head and neck squamous cell carcinoma, in which LncRNAs are becoming new biomarkers in tumor diagnosis.
Disclosure of Invention
The invention aims to provide a diagnostic marker for head and neck cell carcinoma and application thereof, and provides a means for diagnosis or treatment scheme selection or prognosis evaluation of head and neck squamous cell carcinoma so as to find early treatment.
The technical scheme adopted by the invention is as follows:
a diagnostic marker for head and neck cell cancer is LncRNA-RP11-215P8.4, the nucleotide sequence of LncRNA-RP11-215P8.4 is shown as SEQ ID NO. 1, and the length is 669 bp.
Use of LncRNA-RP11-215P8.4 for the preparation of a kit for the diagnostic and/or prognostic evaluation of head and neck squamous cell carcinoma, wherein the nucleotide sequence of LncRNA-RP11-215P8.4 is shown in SEQ ID NO:1 and is 669bp in length.
Further, primers for amplifying the nucleotide sequence of LncRNA-RP11-215P8.4 are shown in SEQ ID NO. 2 and SEQ ID NO. 3.
A reagent kit for detecting the expression quantity of LncRNA-RP11-215P8.4 is an RT-PCR reagent and an RNA detection probe for detecting the expression quantity of LncRNA-RP11-215P 8.4.
Further, the RT-PCR reagent comprises an RNA extraction reagent, a reverse transcription reagent, a PCR quantitative reagent, a forward amplification primer such as SEQ ID NO. 2, a reverse amplification primer such as SEQ ID NO. 3, a cDNA sample used as a negative control and a cDNA sample used as a positive control and highly expressing RP11-215P 8.4.
An agent for preventing and/or treating head and neck squamous cell carcinoma, which comprises an effective active ingredient in an amount that inhibits the expression of LncRNA-RP11-215P8.4 in cancer cells.
Further, the active ingredient for inhibiting the expression amount of LncRNA-RP11-215P8.4 in cancer cells is siRNA for inhibiting the expression of LncRNA-RP11-215P8.4, and the sequence of the siRNA is shown as SEQ ID NO. 4, SEQ ID NO. 5 or SEQ ID NO. 6.
A group of interfering RNAs aiming at RP11-215P8.4 gene targets, wherein the sequence of the RP11-215P8.4 gene targets is shown as SEQ ID NO:1, and the interfering RNAs comprise at least two of siRNA-1, siRNA-2 or siRNA-3, wherein:
the sequence of the siRNA-1 is shown as SEQ ID NO. 4;
the sequence of the siRNA-2 is shown as SEQ ID NO. 5;
the sequence of the siRNA-3 is shown as SEQ ID NO. 6.
The present inventors have found that the expression of LncRNA RP11-215P8.4 in tumors is different from that in normal tissues by analyzing RNA data from SCCHN patients and cancer-free controls. Then, further experiments are carried out, the influence on the migration and invasion of the tumor cells is observed by knocking down the expression of the lncRNA, the influence shows inhibition, and the lncRNA has the potential of being a biomarker in the tumor cells based on the experimental data, so the invention provides the application of the LncRNA RP11-215P8.4 in preparing the diagnosis agent for the squamous cell carcinoma of head and neck.
The present invention relates to an LncRNA marker for detecting squamous cell carcinoma of head and neck. Also relates to a kit for diagnosing the head and neck squamous cell carcinoma, which comprises a reagent for detecting the expression level of the marker; and an agent for treating head and neck squamous cell carcinoma comprising an active ingredient which inhibits the expression amount of LncRNA in cancer cells. For a patient suspected of suffering from head and neck squamous cell carcinoma, the detection reagent provided by the invention can be used for detection so as to facilitate early discovery and early treatment
Drawings
The accompanying drawings, which are included to provide a further understanding of the invention and are incorporated in and constitute a part of this specification, illustrate embodiments of the invention and together with the description serve to explain the invention and do not limit the invention. In the drawings:
FIG. 1 is a statistical graph showing the expression levels of RP11-215P8.4 in 11 normal tissues and 22 SCCHN tissue samples;
FIG. 2 is a scatter plot showing the distribution of the expression levels of RP11-215P8.4 in SCCHN tissues and control normal tissue samples;
FIG. 3 is a statistical graph of the expression levels of RP11-215P8.4 in normal tissue cells (NC) and tumor cell lines (UMSCC, 10B, Tca8113, OSCC-15, HSC-4);
FIG. 4 is a statistical chart of the expression levels of RP11-215P8.4 after transfer of 3 siRNAs into Tca8113, respectively, where the data show: 0.01 < P < 0.05; 0.001 < P < 0.01; p < 0.001;
FIG. 5 is a statistical graph of the expression level of RP11-215P8.4 after transferring 3 siRNAs into HSC-4, respectively, 0.01 < P < 0.05; 0.001 < P < 0.01; p < 0.001;
FIG. 6 is a statistical chart of the expression levels of RP11-215P8.4 after transfer of 3 siRNAs into OSCC-15, respectively, where P is more than 0.01 and less than 0.05; 0.001 < P < 0.01; p < 0.001;
FIG. 7 is a photograph of a scratch test for Tca 8113;
FIG. 8 is a relative migration capability of Tca8113 calculated from the experimental photograph of FIG. 7;
FIG. 9 is a photograph showing the scratch test of HSC-4;
FIG. 10 is a graph of the relative migratory capacity of HSC-4 as calculated from the experimental photograph of FIG. 9;
FIG. 11 is a photograph of a scratch test of OSCC-15;
FIG. 12 is the relative migration capability of OSCC-15 calculated from the experimental photograph of FIG. 11;
FIG. 13 is a photograph of a transwell experiment of Tca8113, HSC-4 and OSCC-15;
FIG. 14 is the relative migratory capacity of Tca8113 calculated from the experimental photograph of FIG. 13;
FIG. 15 is a graph of the relative migratory capacity of HSC-4 as calculated from the experimental photograph of FIG. 13;
FIG. 16 is the relative migration capability of OSCC-15 calculated from the experimental photograph of FIG. 13;
FIG. 17 is a graph of experimental data for the detection of CCK8 according to Tca 8113;
FIG. 18 is a graph plotting experimental data for the detection of CCK8 based on HSC-4;
FIG. 19 is a graph plotting experimental data for the detection of CCK8 based on OSCC-15.
Detailed Description
The following embodiments of the present invention will be described in detail with reference to the accompanying drawings, so that the implementation process of the present invention, which adopts technical means to solve the technical problems and achieve the technical effects, can be fully understood and implemented.
Detection of expression level of LncRNA RP11-215P8.4 in 1.22 SCCHN tissue samples 1.1RNA extraction
(1) Grinding a tissue sample by using liquid nitrogen, adding 1mL of Trizol for cracking, washing a cell sample twice by using PBS (phosphate buffer solution), adding 1mL of Trizol for cracking, transferring the ground tissue or cell into a 1.5mL centrifuge tube without RNase, and standing at room temperature for 7 min;
(2) chloroform was added to 1mL of Trizol in an amount of 200. mu.L of chloroform, vigorously shaken vigorously, and allowed to stand at room temperature for 3 min. Centrifuging at 12000g for 15min at 4 ℃;
(3) taking out the centrifuge tube, dividing the liquid into three layers, taking the upper layer as the target RNA extract, sucking 500 mu L of supernatant liquid by a liquid-moving machine, and carefully transferring the supernatant liquid into a new centrifuge tube;
(4) adding isopropanol into a new centrifuge tube according to the proportion of 1:1, reversing, mixing uniformly, and standing for 10min at room temperature. Centrifuging at high speed for 10min at 4 ℃ and 12000 g;
(5) the supernatant was discarded, and the precipitate was discarded, and washed with 75% ethanol/mL (prepared in 750. mu.L absolute ethanol + 250. mu.L DEPC water). Centrifuging at 4 deg.C for 5min at 8000 g;
(6) repeating the step (5) once or twice;
(7) pouring out supernatant, centrifuging instantly, sucking out residual liquid at the bottom of the centrifuge tube by using a pipettor, drying and precipitating in a fume hood for 3min, adding 20 mu L of NF water to dissolve RNA precipitate after the precipitate is dried, and carefully blowing, beating and mixing uniformly;
(7) mu.L of the RNA solution was taken and the RNA concentration and purity were measured with NanoDrop.
1.2 reverse transcription of RNA
Kang is HiFiScript cDNA Synthesis Kit:
total reaction 20 μ L: 5 × RT Buffer 4 μ L, dNTP Mix 4 μ L, PrimerMix 2 μ L, DTT 2 μ L, HiFiScript 1 μ L, RNA template 1 μ g, NF water to 20 μ L;
1.3qPCR
naoza ChamQ SYBR qPCR Master Mix:
total reaction 20 μ L: 2 × SYBR Green PCR Master mix 10 μ L, upstream primer 0.5 μ L, downstream primer 0.5 μ L, cDNA 1 μ L, NF water 8 μ L;
the primer sequence is as follows:
RP11-215P8.4-F: 5'-ACAGCCCAGACAAACTTCTTCCC-3', shown as SEQ ID NO:2,
RP11-215P8.4-R: 5'-AATGAATGGACCCAGAAGGCAGAC-3', shown as SEQ ID NO: 3.
The amplification reaction was performed using an FTC3000 fluorescent quantitative PCR instrument, programmed as follows: 95-30 s; circulating for 40 seconds at 95-10 seconds and 60-30 seconds; 95-30 s, 60-90 s and 94-10 s.
Wherein the fluorescent signal is collected in 60-30 s.
As shown in FIG. 1, 11 normal tissues and 22 SCCHN confirmed tissues showed that LncRNA RP11-215P8.4 was highly expressed in tumor tissues to different extents, and FIG. 2 showed that LncRNA RP11-215P8.4 was significantly more expressed than normal bladder tissues (P < 0.05), so LncRNA RP11-215P8.4 was considered as one of diagnostic markers for SCCHN.
Detection of expression level of LncRNA RP11-215P8.4 in SCCHN cell line
The expression level of LncRNA RP11-215P8.4 in five SCCHN cell lines, namely UMSCC, 10B, Tca8113, OSCC-15 and HSC-4, was determined by the PCR method and compared with that in normal tissue cells NC.
As a result, as shown in FIG. 3, the expression level of LncRNA RP11-215P8.4 in the five cells was higher than that of normal tissue cell NC, especially the differences among Tca8113, OSCC-15 and HSC-4 were more obvious.
Knockdown of LncRNA RP11-215P8.4
According to the experimental results, Tca8113, OSCC-15 and HSC-4 cell lines are selected to be used for carrying out the knockdown experiment of LncRNA RP11-215P8.4, and the knocked down cells are subjected to the detection of related gene expression and cell migration capacity.
Knockdown of LncRNA RP11-215P8.4 was achieved using siRNA as follows: spreading cells in a six-well plate, and starving for 30min by using Opti-MEM when the culture plate is full of 50% in the next day;
2.5nmol siRNA/siNC was dissolved in 125. mu.L DEPC water, 10. mu.L per well. Taking two tubes as reference numbers 1 and 2, and adding 100 mu L of Opti-MEM into each tube; add 10. mu.L lipofectamine 3000 into 2 tubes, add siRNA or siNC 10. mu.L into 2 tubes, mix evenly and stand for 5min at room temperature. And (3) fully and uniformly mixing the tubes 1 and 2, standing for 15min, adding into a six-hole plate, and transfecting for 6-8 h.
Three siRNAs were used in the study of the present invention, the sequences are as follows:
siRNA-1：5’-AUUGAAGAACAAAAUCCUCCU-3’；
siRNA-2：5’-UGGUUAAUGAAAUUUCAGGGA-3’；
siRNA-3：5’-AGACUUUAUCUUGUGAAUGAU-3’；
FIGS. 4, 5 and 6 show that the expression level of RP11-215P8.4 is reduced in three cell lines after the three siRNAs are transferred into Tca8113, HSC-4 and OSCC-15 cell lines respectively.
3.2 migration Capacity test of tumor cells
Tumor cells were subjected to scarification experiments (fig. 6-12) and transwell experiments (fig. 13-16).
The scratch test is specifically as follows:
after transfection, when the cell density reaches about 70%, quickly marking lines (2 lines for horizontal and vertical lines) in the cell culture plate perpendicular to the pore plate, and ensuring that the width of each scratch is consistent as much as possible. The cell culture medium was discarded and the cells were washed three times with PBS buffer. The medium without serum was added and photographed with an inverted microscope and the photographing time was recorded as 0 h. The cells were placed in a box for culture, and photographs were taken at the same field under an inverted microscope for 24 hours and 48 hours, respectively, and the migration of the cells in the control group and the experimental group was observed.
transwell experiment:
after transfection, cells were digested and centrifuged, cells were resuspended in serum-free medium, and after cell counting, cells were resuspended in serum-free medium at 2X 10 per cell4The number of cells was prepared as a cell suspension. 500 μ L of complete medium containing 10% FBS was added under a Transwell chamber (i.e., the bottom of a 24-well plate), 200 μ L of the formulated cell suspension was added to the chamber, and cultured at 37 ℃ for 36-48h in an incubator containing 5% CO 2. The Transwell chamber was removed and the medium was discarded. Fixing with 4% paraformaldehyde at room temperature for 30 min. Adding 500 mul of 0.1% crystal violet staining solution into each hole, completely immersing the bottom membrane of the transwell cell in the crystal violet staining solution, and staining for 30min at normal temperature. The number of cells passing through the chamber at the bottom of the chamber was observed by taking a picture under an inverted microscope.
As shown in the figure, compared with the SCCHN cells which are not knocked down, the Tca8113, HSC-4 and OSCC-15 cells which are transferred with siRNA have reduced migration capability.
3.3 viability assay of tumor cells
Digesting transfected cells, centrifuging and resuspending cells, counting the number of cells by using a cell counting plate, setting the number of cells in a control group and an experimental group to be 0h, 24h and 48h respectively, and setting the number of cells in three groups, rotating a 96-well plate to 37 ℃, and carrying out cell separation on the cells containing 5% CO2The cells are taken out after being transfected for 0h, 24h and 48h respectively, CCK-8 reagent with the total volume of 10 percent of the culture medium is added into each hole under the condition of keeping out of the sun, the culture is continued for 50min, and the solution in the 96-hole plate is changed from colorless to yellow. The absorbance (OD value) of each well in the 96-well plate was measured at a wavelength of 450nm using a microplate reader.
The results are shown in FIGS. 17-19, where the viability of the tumor cells was significantly reduced.
The results show that inhibitors of expression of LncRNA-RP11-215P8.4 are effective in inhibiting migration of SCCHN cells overexpressing RP11-215P8.4, and thus can be used as therapeutic or adjunctive therapeutic agents for such SCCHN.
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, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Although the embodiments of the present invention have been described in conjunction with the accompanying drawings, those skilled in the art can make various modifications and variations without departing from the spirit and scope of the invention, and such modifications and variations fall within the scope defined by the appended claims.
<110> Shanxi Saishang Biotechnology Co., Ltd
<120> diagnostic marker for head and neck cell cancer and application thereof
<130> 1
<160> 6
<170> PatentIn version 3.5
<210> 1
<211> 669
<212> DNA
<213> Artificial Sequence
<400> 1
GCAATGGCTG AAAATGGAGA TGAAGAGTTT CACTCTCAGA GGAGGATTTT GTTCTTCAAT 60
TGTGGAGTGA TCTCTATCAC CAGTGACTAA AGATGTTGGA GCACAGAGAG CCATACCCCA 120
AAATATGATG CTTCGGCATG CTGACTGCTT TGAAAATTGA AAGGCCTCAG AAATAATCCT 180
CAGTGCCAGG GTCTCCCTCT GACCTCCCCC TACCTCCCTT TCTCTCTGAT CCTGTCTCTC 240
CCAAAGCACA GAATGAAGCT GTTCTCTGAA TTCCCTTATC TACCTAGAAA CTGGACCCCC 300
AAAGAGGAAC ACAATTTGCC TTTGATCCCT TCCCTGAAAT TTCATTAACC AGAGAAAATT 360
AAAACTTCTA TCACAAAGGA AGAGACTGAA CATTAAACAC CATAGCTACA GCCCAGACAA 420
ACTTCTTCCC AAACCATTGT TTGTTCTCCT GCCTGTTAAA TTGCCAGAGA ATCATTCACA 480
AGATAAAGTC TGCCTTCTGG GTCCATTCAT TCCCCACTAA AAATCTTTTA CTCCTACACC 540
CTCATGTCTC CTTCCTCCAT GAAGAAGGGT CTATAAACCT CTAGGCCTCA TTGGGTTATT 600
GGGTAATCAT TCTCATGCAG TTCCCCTGTG CTCTGCATGT TAAATAAATT TGTATGCCTT 660
TTTCTCCTA 669
<210> 2
<211> 23
<212> DNA
<213> Artificial Sequence
<400> 2
ACAGCCCAGA CAAACTTCTT CCC 23
<210> 3
<211> 24
<212> DNA
<213> Artificial Sequence
<400> 3
AATGAATGGA CCCAGAAGGC AGAC 24
<210> 4
<211> 21
<212> RNA
<213> Artificial Sequence
<400> 4
AUUGAAGAAC AAAAUCCUCC U 21
<210> 5
<211> 21
<212> RNA
<213> Artificial Sequence
<400> 5
UGGUUAAUGA AAUUUCAGGG A 21
<210> 6
<211> 21
<212> RNA
<213> Artificial Sequence
<400> 6
AGACUUUAUC UUGUGAAUGA U 21
Claims (4)
1. The application of a reagent for detecting LncRNA-RP11-215P8.4 in preparing a kit for diagnosing head and neck squamous cell carcinoma is disclosed, wherein the nucleotide sequence of LncRNA-RP11-215P8.4 is shown as SEQ ID NO. 1, and the length is 669 bp;
the head and neck squamous cell carcinoma is tongue carcinoma or oral cancer.
2. The use according to claim 1, wherein the primers for amplifying the nucleotide sequence of LncRNA-RP11-215P8.4 are represented by SEQ ID NOs 2 and 3.
3. Use of an agent for inhibiting the expression level of LncRNA-RP11-215P8.4 for the preparation of a medicament for the treatment of head and neck squamous cell carcinoma, wherein said agent comprises an active ingredient in an amount effective to inhibit the expression level of LncRNA-RP11-215P8.4 in cancer cells;
the head and neck squamous cell carcinoma is oral cancer or tongue cancer.
4. The use according to claim 3, wherein the active ingredient for inhibiting the expression amount of LncRNA-RP11-215P8.4 in cancer cells is siRNA having the sequence shown in SEQ ID NO. 4, SEQ ID NO. 5 or SEQ ID NO. 6, which inhibits the expression of LncRNA-RP11-215P 8.4.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010075552.4A CN113151456B (en) | 2020-01-22 | 2020-01-22 | Diagnostic marker for head and neck cell carcinoma and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010075552.4A CN113151456B (en) | 2020-01-22 | 2020-01-22 | Diagnostic marker for head and neck cell carcinoma and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN113151456A CN113151456A (en) | 2021-07-23 |
| CN113151456B true CN113151456B (en) | 2022-07-22 |
Family
ID=76881844
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202010075552.4A Active CN113151456B (en) | 2020-01-22 | 2020-01-22 | Diagnostic marker for head and neck cell carcinoma and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113151456B (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108823308A (en) * | 2018-05-28 | 2018-11-16 | 中南大学 | Detect application and the kit of circMAN1A2 and LOC284454 reagent |
| KR20190048019A (en) * | 2017-10-30 | 2019-05-09 | 이화여자대학교 산학협력단 | Compositions comprising lncRNA for diagnosis and prognosis of epithelial-mesenchymal transition in lung cancer |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20140329704A1 (en) * | 2013-03-28 | 2014-11-06 | President And Fellows Of Harvard College | Markers for mature beta-cells and methods of using the same |
-
2020
- 2020-01-22 CN CN202010075552.4A patent/CN113151456B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20190048019A (en) * | 2017-10-30 | 2019-05-09 | 이화여자대학교 산학협력단 | Compositions comprising lncRNA for diagnosis and prognosis of epithelial-mesenchymal transition in lung cancer |
| CN108823308A (en) * | 2018-05-28 | 2018-11-16 | 中南大学 | Detect application and the kit of circMAN1A2 and LOC284454 reagent |
Non-Patent Citations (4)
| Title |
|---|
| "LncRNA LINC00460 promotes EMT in head and neck squamous cell carcinoma by facilitating peroxiredoxin-1 into the nucleus";Yingying Jiang等;《Journal of Experimental & Clinical Cancer Research 》;20190820;第38卷;第1-19页 * |
| "Transcript: lnc-CDKN2AIPNL-1:1";LNCipedia;《LNCipedia》;20180430;第1-2页 * |
| "长链非编码RNA HOXA11-AS表达对头颈部鳞状细胞癌细胞迁移和患者预后的影响";张莉等;《新乡医学院学报》;20180430;第35卷(第4期);第310-313页 * |
| "长链非编码RNA在口腔鳞状细胞癌中的研究进展";黄佳欣等;《分子诊断与治疗杂志》;20180331;第10卷(第2期);第120-124页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN113151456A (en) | 2021-07-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109371131B (en) | Molecular marker LncRNA DANCR for diagnosing and treating bladder cancer and application thereof | |
| CN106701900B (en) | Long-chain non-coding RNA HERC2P3 gene and application thereof in gastric cancer | |
| Ma et al. | LncRNA FER1L4 suppressed cancer cell growth and invasion in esophageal squamous cell carcinoma. | |
| CN111118156B (en) | Molecular marker LncRNA AC012640.1 for diagnosing and treating bladder cancer and application thereof | |
| CN111518909B (en) | Application of METTL2 gene in preparation of kit for detecting treatment sensitivity of colorectal cancer fluorouracil drugs | |
| CN107164554B (en) | Application of ASPRV1 as biomarker in diagnosis and treatment of laryngeal squamous cell carcinoma | |
| CN108034655B (en) | Application of long non-coding RNA and composition thereof in diagnosis/treatment of colorectal cancer | |
| CN113151456B (en) | Diagnostic marker for head and neck cell carcinoma and application thereof | |
| CN107312855A (en) | A kind of gene related to larynx squamous carcinoma and its application | |
| CN106995857B (en) | Application of biomarker ENSG00000267416 in cancer | |
| CN112641797B (en) | Target and diagnostic marker for inhibiting colorectal cancer growth and metastasis and application thereof | |
| CN112430663B (en) | Biomarker for diagnosis and treatment of bladder cancer and application thereof | |
| CN111424082A (en) | Application of lncRNA-SNHG6 gene in preparation of medicine for treating osteosarcoma | |
| CN110607368B (en) | Application of miRNA3926-1 gene as pancreatic cancer diagnosis and curative effect marker | |
| CN115779090B (en) | Application of substance for improving UBE2J1 expression in preparation of anticancer drugs | |
| AU2020342299B2 (en) | Tumour marker aquaporin 2 protein and application thereof | |
| CN114917348B (en) | Application of nerve adhesion factor in preparation of reagent for inhibiting and marking liver cancer cell metastasis | |
| CN114561466B (en) | Application of FTO in diagnosis and treatment of osteosarcoma | |
| CN108165624A (en) | Application of the biomarker in osteosarcoma diagnosis and treatment | |
| CN116284315B (en) | SSX polypeptide and application thereof in treating synovial sarcoma | |
| LU503535B1 (en) | Use of linc02539 in diagnosis and treatment of kidney cancer | |
| CN109055551B (en) | Application of miR-3687 as target for treating bladder cancer | |
| CN114177192A (en) | Target for inhibiting colorectal cancer proliferation and application thereof | |
| Bai et al. | Long Intergenic Non-Protein Coding RNA 312 Regulates FRABIN to Inhibit the Occurrence and Development of Pancreatic Cancer | |
| Hei et al. | MicroRNA-370 Targets Forkhead Box M1 Regulates Migration and Invasion in Colorectal Cancer Cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20220926 Address after: 710054 Factory 413, Block C, Boyuan Science and Technology Plaza, Xi'an Jiaotong University Science and Technology Park, No. 99 Yanxiang Road, Yanta District, Xi'an City, Shaanxi Province Patentee after: Xi'an Aorui Jingchuang Biotechnology Co.,Ltd. Address before: Room 413, block C, Boyuan science and Technology Plaza, Xi'an Jiaotong University Science and Technology Park, 99 Yanxiang Road, Yanta District, Xi'an City, Shaanxi Province, 710065 Patentee before: SHAANXI FAN CHANG BIOTECHNOLOGY Co.,Ltd. |