CN112877434A - Group of circRNA markers and primers for detecting esophageal cancer tissues, application of primers and kit containing primers - Google Patents

Group of circRNA markers and primers for detecting esophageal cancer tissues, application of primers and kit containing primers Download PDF

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CN112877434A
CN112877434A CN202110198287.3A CN202110198287A CN112877434A CN 112877434 A CN112877434 A CN 112877434A CN 202110198287 A CN202110198287 A CN 202110198287A CN 112877434 A CN112877434 A CN 112877434A
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hsa
circ
primers
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冯刚
刘康
张若兰
别俊
熊蓉
胡欣
刘建林
刘燕群
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Nanchong Central Hospital
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Abstract

The invention relates to a group of c I rcRNA markers for detecting esophageal cancer tissues, which comprise one of the following three c I rcRNA markers, wherein the three c I rcRNA markers are respectively shown as SEQ ID No: 1, hsa _ c I rc _0004390, and the ribonucleic acid sequence shown in SEQ I D No: 4 and the sequence of the ribonucleic acid and hsa _ c I rc _0060927 shown in SEQ I D No: hsa _ c i rc _0111350 shown in FIG. 7. The invention has the beneficial effects that a group of special ci rcRNA markers are provided for the first time and can be used as the c i rcRNA markers of esophageal squamous carcinoma tissues; the high expression of the marker in esophageal squamous carcinoma is verified, so that the marker can be applied to products for detecting esophageal squamous carcinoma tissues; the kit contains a specific primer aiming at any one biomarker, and can be used for detecting esophageal squamous cell carcinoma tissues, so that the detection process is quicker and more accurate.

Description

Group of circRNA markers and primers for detecting esophageal cancer tissues, application of primers and kit containing primers
Technical Field
The invention relates to the technical field of gene detection, in particular to a group of circRNA markers and primers for detecting esophageal cancer tissues, application of the circRNA markers and the primers and a kit containing the circRNA markers and the primers.
Background
Circular RNA (circRNA) is a special non-coding RNA molecule, sometimes expressed in living body, and is the latest research hotspot in the RNA field.
Unlike traditional linear RNA (containing 5 'and 3' ends), the circRNA molecule is in a closed ring structure, is not influenced by RNA exonuclease, is more stable in expression and is not easy to degrade. Functionally, recent researches show that circRNA molecules are rich in microrna (miRNA) binding sites and play a role of miRNA sponge (miRNA sponge) in cells, so that the inhibition effect of miRNA on target genes of the cells is relieved, and the expression level of the target genes is increased; this mechanism of action is known as the competitive endogenous rna (cerna) mechanism. circRNA plays an important regulatory role in disease through the interaction with disease-associated mirnas.
Esophageal cancer is a (esophageal cancer) common malignant tumor in human, accounts for more than 90% of esophageal tumors, and is second only to gastric cancer in the review survey of all malignant tumor deaths, and is the 2 nd place. It is estimated that about 20 million people die of esophageal cancer every year worldwide, one of the most common malignancies with great harm to people's life and health.
Esophageal cancer is classified into various types, including esophageal squamous carcinoma, esophageal adenocarcinoma, esophageal stromal tumor, esophageal sarcoma, esophageal mucinous adenocarcinoma. In the initial cases, the domestic esophageal squamous cell carcinoma accounts for more than 90% of all esophageal cancers. Different treatment regimens for esophageal cancer are different, and therefore, rapid and accurate detection of esophageal squamous carcinoma tissue is a very important technology.
In the prior art, for determining the type of esophageal cancer, a needle biopsy is often needed, so that the method is not only inefficient, but also the accuracy of detection is influenced in various aspects due to different actual conditions.
In the prior art, there is a circRNA screening which relates to high expression of esophageal squamous cell carcinoma, but no disclosure is provided for screening out a determined circRNA sequence. At present, the high-expression circRNA screening of esophageal squamous carcinoma is mainly performed through second-generation sequencing, and a great amount of confidence data analysis is required to be performed on a sequencing result, wherein errors may exist.
Disclosure of Invention
The invention aims to solve the technical problem of providing a group of circRNA markers and primers for detecting esophageal cancer tissues, application thereof and a kit containing the same.
The technical scheme for solving the technical problems is as follows: a group of circRNA markers for detecting esophageal cancer tissues comprises one of the following three circRNA markers, wherein the three circRNA markers are respectively shown as SEQ ID No: 1, hsa _ circ _0004390, ribonucleic acid sequence shown in SEQ ID No: 4 and the sequence of the ribonucleic acid and hsa _ circ _0060927 shown in SEQ ID No: hsa _ circ _0111350 shown in FIG. 7.
Further, the esophageal cancer tissue is esophageal squamous carcinoma tissue.
The invention provides a primer of the circRNA marker for detecting esophageal cancer tissues, which is characterized by comprising at least one specific primer aiming at any circRNA marker.
The invention provides a primer of the circRNA marker for detecting esophageal cancer tissues, wherein one primer is a specific primer aiming at the hsa _ circ _0004390 marker and comprises the primer shown as SEQ ID No: 2 and the upstream primer shown as SEQ ID No: 3, or a reverse primer as shown in the figure.
The invention provides a primer of the circRNA marker for detecting esophageal cancer tissues, wherein one primer is a specific primer aiming at the hsa _ circ _0060927 marker and comprises the primer shown as SEQ ID No: 5 and the upstream primer shown as SEQ ID No: 6.
The invention provides a primer of the circRNA marker for detecting esophageal cancer tissues, wherein one primer is a specific primer aiming at the hsa _ circ _0111350 marker and comprises the primer shown as SEQ ID No: 8 and the upstream primer shown as SEQ ID No: 9 as shown in the figure.
The invention provides application of the primer of the circRNA marker for detecting the esophageal cancer tissue in preparation of a kit for detecting the esophageal cancer tissue.
The invention provides a kit for detecting esophageal cancer tissues, which comprises the specific primer.
The invention has the beneficial effects that:
1) the invention provides hsa _ circ _0004390, hsa _ circ _0060927 and hsa _ circ _0111350 for the first time as circRNA markers of esophageal squamous carcinoma tissues;
2) the invention verifies the high expression of hsa _ circ _0004390, hsa _ circ _0060927 and hsa _ circ _0111350 in esophageal squamous cell carcinoma, so that the kit can be applied to products for detecting esophageal squamous cell carcinoma tissues;
3) the invention provides a kit, which contains specific primers aiming at any one of the markers hsa _ circ _0004390, hsa _ circ _0060927 and hsa _ circ _0111350, and can make the detection process faster and more accurate.
Drawings
FIG. 1 is a comparison of the sequencing CPM values of hsa _ circ _0004390 in esophageal squamous cell carcinoma tissue and paracancerous tissue in example 1 of the present invention, wherein Nomal is paracancerous tissue and ESCC is esophageal squamous cell carcinoma tissue;
FIG. 2 is a graph showing the relative expression level of has-circ-0004390 in esophageal squamous carcinoma tissue and paracarcinoma tissue in example 1 of the present invention, wherein the ordinate is a multiple.
FIG. 3 is a comparison of the sequencing CPM values of hsa _ circ _0060927 in esophageal squamous cell carcinoma tissue and paracancerous tissue in example 1 of the present invention, wherein Nomal is paracancerous tissue and ESCC is esophageal squamous cell carcinoma tissue;
FIG. 4 shows the relative expression level of hsa _ circ _0060927 in esophageal squamous carcinoma tissue and paracarcinoma tissue in example 1 of the present invention, wherein the ordinate is fold.
FIG. 5 is a comparison of the sequencing CPM values of hsa _ circ _0111350 in esophageal squamous cell carcinoma tissue and paracancerous tissue in example 1 of the present invention, wherein Nomal is paracancerous tissue and ESCC is esophageal squamous cell carcinoma tissue;
FIG. 6 shows the relative expression level of hsa _ circ _0111350 in esophageal squamous carcinoma tissue and paracarcinoma tissue in example 1 of the present invention, wherein the ordinate is fold.
Detailed Description
The principles and features of this invention are described below in conjunction with the following drawings, which are set forth by way of illustration only and are not intended to limit the scope of the invention.
The ribonucleic acid sequence of hsa _ circ _0004390 of the invention is shown in SEQ ID No: 1 (ggcccccatt tacagcagga gttaccttgt tttctggaca gtgtccaacc tcatggcctt cctcatcatg gttgtggtgt acctgcggat ctacgtgtac gtcaagagga aaaccaacgt cttgtctccg catacaagtg ggtccatcag ccgccggagg acacccatga agctaatgaa gacggtgatg actgtcttag gatgttcact tctccacaat gaatgagtgt cactatgaca agcacatgga ctttttttat aataggagca acactgatac tgtcgatgac tggacaggaa caaagcttgt gattgttttg tgtgttggga cgtttttctg cctgtttatt tttttttcta attctctggt catcgcggca gtgatcaaaa acagaaaatt tcatttcccc ttctactacc tgttggctaa tttagctgct gccgatttct tcgctggaat tgcctatgta ttcctgatgt ttaacacagg cccagtttca aaaactttga ctgtcaaccg ctggtttctc cgtcaggggc ttctggacag tagcttgact gcttccctca ccaacttgct ggttatcgcc gtggagaggc acatgtcaat catgaggatg cgggtccata gcaacctgac caaaaagagg gtgacactgc tcattttgct tgtctgggcc atcgccattt ttatgggggc ggtccccaca ctgggctgga attgcctctg caacatctct gcctgctctt ccct); the ribonucleic acid sequence of hsa _ circ _0060927 is shown in SEQ ID No: 4 (tcttccattt ggcgttggaa aaagaatgtg cattggtcgc cgattagcag agcttcaact gcatttggct ctttgttgga ttgtccgcaa atacgacatc caggccacag acaatgagcc tgttgagatg ctacactcag gcaccctggt gcccagccgg gaactcccca tcgcgttttg ccagcgataa tacgcctcag ggaaggggaa gactggcagc gggtccggag tgcctttcaa aagaaactaa tgaaaccagg ggaagtgatg aagctggaca acaaaatcaa tgaggtcttg gccgatttta tgggcagaat agatgagctc tgtgatgaaa gaggccacgt tgaagacttg tacagcgaac tgaacaaatg gtcgtttgaa agtatctgcc tcgtgttgta tgagaagaga tttgggcttc tccagaagaa tgcaggggat gaagctgtga acttcatcat ggccatcaaa acaatgatga gcacgtttgg gaggatgatg gtcactccag tcgagctgca caagagcctc aacaccaagg tctggcagga ccacactctg gcctgggaca ccattttcaa atcagtcaaa gcttgtatcg acaaccggtt agagaagtat tctcagcagc ctagtgcaga tttcctttgt gacatttatc accagaatcg gctttcaaag aaagaattgt atgctgctgt cacagagctc cagctggctg cggtggaaac gacagcaaac agtctaatgt ggattctcta caatttatcc cgtaatcccc aagtgcaaca aaagcttctt aaggaaattc aaagtgtatt acctgagaat caggtgccac gggcagaaga tttgaggaat atgccgtatt taaaagcctg tctgaaagaa tctatgaggc ttacgccgag tgtaccattt acaactcgga ctcttgacaa ggcaacagtt ctgggtgaat atgctttacc caaaggaaca gtgctcatgc taaataccca ggtgttggga tccagtgaag acaattttga agattcaagt cagtttagac ctgaacgttg gcttcaggag aaggaaaaaa ttaatccttt tgcgca); the ribonucleic acid sequence of hsa _ circ _0111350 is shown in SEQ ID No: shown at 7 (aaatggttct tcagattctt ccacactgcc caaattggaa gaattctgta cctctcttac ccagtcagag cagtcagcag acgggagcca gtctgaaccc aacaacagtc agactcagcc aaagcaaatt cagctttcca cagcagcacc ctgttcaaca actgcagtgg atgattccgc agaaaagccc tctggttctg gatgagatcc ccactggaat gccatgcctg gagtcagtga ccataggtga tgatatttgg gatgagaact ggttacctct acaggctaag atgggaaaag gaggtgatgc tgcctcccat ctatttactg caagccttgg tggaaagaat cagtattcat catgtaaaga aatgccacag aaggactggt gtttttctac ccctaaagat acatgggatg attcttggca gccttcaggc cttgtaaatg gaacgaaagt agaagttcat aagccagaag tactgggtgc tcaggaaaaa aatactggca caaacaggac tcaaaagcaa ctagacat).
As shown in figure 1, the comparison of the CPM value of hsa _ circ _0004390 sequencing in the esophageal squamous carcinoma cancer tissue and the adjacent cancer tissue shows that the expression level of the CPM value in the adjacent cancer tissue is very low and is basically 0, while the expression level in the esophageal squamous carcinoma cancer tissue is significantly higher, and the CPM value is in the range of 0-4000.
As shown in FIG. 3, the comparison of the CPM value of hsa _ circ _0060927 sequencing in the esophageal squamous carcinoma cancer tissue and the paracancerous tissue shows that the expression level of the CPM value in the paracancerous tissue is very low and is basically 0, while the expression level of the CPM value in the esophageal squamous carcinoma cancer tissue is significantly higher and is in the range of 0-20000.
As shown in FIG. 5, the comparison of the CPM value of hsa _ circ _0111350 sequencing in the esophageal squamous carcinoma cancer tissue and the paracancerous tissue shows that the expression level of the CPM value in the paracancerous tissue is very low and is basically 0, while the expression level of the CPM value in the esophageal squamous carcinoma cancer tissue is significantly higher and the CPM value is in the range of 0-620.
Preferably, the esophageal cancer tissue is esophageal squamous carcinoma tissue.
Preferably, the specific primers comprise:
specific primers for hsa _ circ _0004390, as shown in SEQ ID No: 2 (tgtctccgca tacaagtggg) as shown in SEQ ID No: 3 (aagctttgtt cctgtccagt ca).
Specific primers for hsa _ circ _0060927, as shown in SEQ ID No: 5 (gagcttcaac tgcatttggc t) as shown in SEQ ID No: 6 (ttgaaaggca ctccggacc).
Specific primers for hsa _ circ _0111350, as shown in SEQ ID No: 8, as shown in SEQ ID No: 9 downstream primer (tggtcactga ctccaggcat)
The specific primers of any group of circRNA markers for detecting the esophageal cancer tissues are applied to the preparation of the kit for detecting the esophageal cancer tissues.
The kit for detecting esophageal cancer tissues comprises any one group of specific primers.
The following examples are the verification experiments that the above three markers can be used as circRNA markers for esophageal squamous carcinoma tissue detection.
Example 1
1) Collecting a sample: tissue samples of 6 patients with esophageal squamous carcinoma who underwent surgery at the central hospital of Nanxi city were collected, and the corresponding paracarcinoma tissues of the patients were used as a control group. The pathological diagnosis of the patient is verified by the pathologist. Patients have not received radiotherapy or chemotherapy before operation. And (3) obtaining a fresh tissue specimen under an aseptic condition, washing blood with physiological saline, subpackaging the washed blood into RNA enzyme-removed cryopreservation tubes, storing the frozen tube in a liquid nitrogen tank, performing high-throughput sequencing, and screening hsa _ circ _0004390, hsa _ circ _0060927 and hsa _ circ _0111350 respectively.
2) Tissue total RNA extraction: shearing the tissues of each experimental group collected in the step 1) on ice, taking 50-100mg of tissues into an EP tube without RNA enzyme, adding lml precooled Trizol, uniformly blowing and beating, and standing for 20min at-20 ℃; adding 200 μ l chloroform, shaking vigorously and mixing well for 30s to make the water phase and organic phase contact sufficiently, standing at room temperature for 5 min; centrifuging at 14000g for 15min at 4 deg.C to obtain three layers, transferring RNA to another new RNase free EP tube at the upper water phase, and sucking 450 ul; adding isovolumetric isopropanol, gently and fully mixing uniformly, and standing at room temperature for 10 min;
in the above steps, the step of mixing thoroughly, specifically, mixing without using an oscillator, and reversing for 6-8 times.
Centrifuging at 4 deg.C for 10min at 14,000g, and removing supernatant; washing twice with 1ml of 75% ethanol, and centrifuging at 12,000g at 4 ℃ for 3 min; after adding the ethanol, only the EP pipe needs to be slightly reversed, the oscillation of an oscillator or the suction and beating of a gun head are not needed for precipitation, and after the supernatant is poured, the residual super clean bench is sucked away by the gun head for air drying; adding a proper amount of DEPC water to dissolve the precipitate. The RNA concentration was measured spectrophotometrically and the RNA solubility and A260/A280 ratio were recorded, the ratio being between 1.8 and 2.1.
3) Designing a primer: as a circular closed structure of circRNA, a back-to-back primer (divergent primer) is designed, and GAPDH is used as an internal reference and sent to a primer synthesis company for synthesis; the sequence of each primer group is as follows:
specific primers for hsa _ circ _ 0004390:
an upstream primer: 5'-TGTCTCCGCATACAAGTGGG-3', respectively;
a downstream primer 5'-AAGCTTTGTTCCTGTCCAGTCA-3';
target gene amplification length: 186 bp.
Specific primers for hsa _ circ _ 0060927:
an upstream primer: 5'-GAGCTTCAACTGCATTTGGCT-3', respectively;
a downstream primer 5'-TTGAAAGGCACTCCGGACC-3';
target gene amplification length: 191 bp.
Specific primers for hsa _ circ _ 0111350:
an upstream primer: 5'-AGCCAGTCTGAACCCAACAA-3', respectively;
a downstream primer 5'-TGGTCACTGACTCCAGGCAT-3';
target gene amplification length: 158 bp.
4) Reverse transcription: the reverse transcription reaction system was configured according to the HiScript 1st Strand cDNA Synthesis Kit (+ gDNA wiper) instructions. The reagents were first subjected to flash dissociation, added to an EP tube with rnase removed using a pipette in sequence, mixed well, placed in a circulator and incubated to reverse RNA to cDNA, reverse transcribed at 37 ℃ for 15min, 85 ℃ for 5sec, and finally stored at 4 ℃. The transcribed cDNA was diluted 10-fold for use.
5) Reverse transcription-polymerase chain reaction (RT-PCR) verified the expression of hsa _ circ _0004390, hsa _ circ _0060927, and hsa _ circ _0111350, respectively, in the samples:
the RT-PCR reaction system was configured as shown in Table 1:
TABLE 1RT-PCR reaction System
Components Volume of
SYBR Premix EX Taq 10ul
Upstream primer 0.5ul
Downstream primer 0.5ul
DNA template 2ul
DdH2O 7ul
The reaction procedure of RT-PCR is denaturation at 95 ℃ for 30 sec; amplification was carried out at 95 ℃ for 5sec, at 58 ℃ for 30sec, for 35 cycles.
6) Statistical analysis
Statistical analysis of experimental data was performed using SPSS17.0 software. Data are expressed as mean ± standard deviation. P less than 0.05 represents a statistical significance of the difference, and all experiments were repeated three times.
The real-time quantitative PCR amplification curve has good overall parallelism, which indicates that the amplification efficiency of each group of reaction tubes is similar, the inflection point of the amplification curve is clear, the limit is flat without raising, the slope of the exponential phase of the curve is larger, and the amplification efficiency is higher; the dissolution curve of the sample amplification product is a single peak, which indicates that the amplification product is only specific amplification.
Relative quantitative formula for RT-PCR: 2-. DELTA.Ct.times.100%.
The results show that the expression level of hsa _ circ _0004390 in esophageal squamous carcinoma tissues is obviously higher than that in paracarcinoma tissues, the difference is statistically significant (P <0.001), and the specific difference of the expression level is 78.53 times that in paracarcinoma tissues, and the specific results are shown in FIG. 2. In FIG. 2, the expression level of hsa _ circ _0004390 in the para-carcinoma tissue is 1 fold, compared with the expression level in the esophageal squamous cell carcinoma tissue, it can be seen from the results that hsa _ circ _0004390 is a good circRNA marker for esophageal squamous cell carcinoma, and can be applied in various aspects.
The results show that the expression level of hsa _ circ _0060927 in esophageal squamous carcinoma tissues is obviously higher than that in paracarcinoma tissues, the difference is statistically significant (P <0.001), and the specific difference of the expression level is 82 times that in paracarcinoma tissues, and the specific results are shown in FIG. 4. In FIG. 4, the expression level of hsa _ circ _0060927 in the para-carcinoma tissue is 1 fold, compared with the expression level in the esophageal squamous cell carcinoma tissue, it can be seen from the results that hsa _ circ _0060927 is a good circRNA marker for esophageal squamous cell carcinoma, and can be applied in various aspects.
The results show that the expression level of hsa _ circ _0111350 in esophageal squamous carcinoma tissues is obviously higher than that in paracarcinoma tissues, the difference is statistically significant (P <0.001), and the specific difference of the expression level is 312 times that in paracarcinoma tissues, and the specific results are shown in FIG. 6. In FIG. 6, the expression level of hsa _ circ _0111350 in the para-carcinoma tissue is 1 fold, compared with the expression level in the esophageal squamous cell carcinoma tissue, it can be seen from the results that hsa _ circ _0111350 is a good circRNA marker for esophageal squamous cell carcinoma, and can be applied in various aspects.
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.
Sequence listing
<110> central hospital of south China Mobile City
<120> a group of circRNA markers for detecting esophageal cancer tissues, primers, application thereof and kit containing the same
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cttgtctccg catacaagtg ggtccatcag ccgccggagg acacccatga agctaatgaa 180
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caaagcttgt gattgttttg tgtgttggga cgtttttctg cctgtttatt tttttttcta 360
attctctggt catcgcggca gtgatcaaaa acagaaaatt tcatttcccc ttctactacc 420
tgttggctaa tttagctgct gccgatttct tcgctggaat tgcctatgta ttcctgatgt 480
ttaacacagg cccagtttca aaaactttga ctgtcaaccg ctggtttctc cgtcaggggc 540
ttctggacag tagcttgact gcttccctca ccaacttgct ggttatcgcc gtggagaggc 600
acatgtcaat catgaggatg cgggtccata gcaacctgac caaaaagagg gtgacactgc 660
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tgttgagatg ctacactcag gcaccctggt gcccagccgg gaactcccca tcgcgttttg 180
ccagcgataa tacgcctcag ggaaggggaa gactggcagc gggtccggag tgcctttcaa 240
aagaaactaa tgaaaccagg ggaagtgatg aagctggaca acaaaatcaa tgaggtcttg 300
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cgtaatcccc aagtgcaaca aaagcttctt aaggaaattc aaagtgtatt acctgagaat 840
caggtgccac gggcagaaga tttgaggaat atgccgtatt taaaagcctg tctgaaagaa 900
tctatgaggc ttacgccgag tgtaccattt acaactcgga ctcttgacaa ggcaacagtt 960
ctgggtgaat atgctttacc caaaggaaca gtgctcatgc taaataccca ggtgttggga 1020
tccagtgaag acaattttga agattcaagt cagtttagac ctgaacgttg gcttcaggag 1080
aaggaaaaaa ttaatccttt tgcgca 1106
<210> 5
<211> 21
<212> DNA/RNA
<213> Artificial Sequence
<400> 5
gagcttcaac tgcatttggc t 21
<210> 6
<211> 19
<212> DNA/RNA
<213> Artificial Sequence
<400> 6
ttgaaaggca ctccggacc 19
<210> 7
<211> 538
<212> DNA/RNA
<213> Homo sapiens
<400> 7
aaatggttct tcagattctt ccacactgcc caaattggaa gaattctgta cctctcttac 60
ccagtcagag cagtcagcag acgggagcca gtctgaaccc aacaacagtc agactcagcc 120
aaagcaaatt cagctttcca cagcagcacc ctgttcaaca actgcagtgg atgattccgc 180
agaaaagccc tctggttctg gatgagatcc ccactggaat gccatgcctg gagtcagtga 240
ccataggtga tgatatttgg gatgagaact ggttacctct acaggctaag atgggaaaag 300
gaggtgatgc tgcctcccat ctatttactg caagccttgg tggaaagaat cagtattcat 360
catgtaaaga aatgccacag aaggactggt gtttttctac ccctaaagat acatgggatg 420
attcttggca gccttcaggc cttgtaaatg gaacgaaagt agaagttcat aagccagaag 480
tactgggtgc tcaggaaaaa aatactggca caaacaggac tcaaaagcaa ctagacat 538
<210> 8
<211> 20
<212> DNA/RNA
<213> Artificial Sequence
<400> 8
agccagtctg aacccaacaa 20
<210> 9
<211> 20
<212> DNA/RNA
<213> Artificial Sequence
<400> 9
tggtcactga ctccaggcat 20

Claims (8)

1. A group of circRNA markers for detecting esophageal cancer tissues is characterized by comprising one of the following three circRNA markers, wherein the three circRNA markers are respectively shown as SEQ ID No: 1, hsa _ circ _0004390, ribonucleic acid sequence shown in SEQ ID No: 4 and the sequence of the ribonucleic acid and hsa _ circ _0060927 shown in SEQ ID No: hsa _ circ _0111350 shown in FIG. 7.
2. The set of circRNA markers for detecting esophageal cancer tissue according to claim 1, wherein the esophageal cancer tissue is esophageal squamous carcinoma tissue.
3. A primer for detecting a circRNA marker of esophageal cancer tissue according to any one of claims 1 or 2, comprising at least one specific primer for any one of the circRNA markers.
4. The primer for detecting the circRNA marker of esophageal cancer tissue according to claim 3, wherein one of the primers is a specific primer for the hsa _ circ _0004390 marker, and comprises the nucleotide sequence shown in SEQ ID No: 2 and the upstream primer shown as SEQ ID No: 3, or a reverse primer as shown in the figure.
5. The primer for detecting the circRNA marker of esophageal cancer tissue according to claim 3, wherein one of the primers is a specific primer for the hsa _ circ _0060927 marker, and comprises the nucleotide sequence shown in SEQ ID No: 5 and the upstream primer shown as SEQ ID No: 6.
6. The primer for detecting the circRNA marker of esophageal cancer tissue according to claim 3, wherein one of the primers is a specific primer for the hsa _ circ _0111350 marker, and comprises the nucleotide sequence shown in SEQ ID No: 8 and the upstream primer shown as SEQ ID No: 9 as shown in the figure.
7. Use of the primer for detecting the circRNA marker of esophageal cancer tissue according to any one of claims 3-6 in the preparation of a kit for detecting esophageal cancer tissue.
8. A kit for detecting esophageal cancer tissues, which comprises the specific primer according to any one of claims 3-6.
CN202110198287.3A 2021-02-22 2021-02-22 Group of circRNA markers and primers for detecting esophageal cancer tissues, application of primers and kit containing primers Pending CN112877434A (en)

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Application publication date: 20210601