CN107475258A - A kind of RNAcircEPSTI1 and its application in three cloudy breast cancer - Google Patents
A kind of RNAcircEPSTI1 and its application in three cloudy breast cancer Download PDFInfo
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- CN107475258A CN107475258A CN201710721878.8A CN201710721878A CN107475258A CN 107475258 A CN107475258 A CN 107475258A CN 201710721878 A CN201710721878 A CN 201710721878A CN 107475258 A CN107475258 A CN 107475258A
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Abstract
The present invention relates to breast cancer diagnosis technical field, specifically discloses a kind of RNA circEPSTI1 and its application in three cloudy breast cancer；The present invention first collects doubtful three cloudy breast cancer tissue's sample to be measured, extracted total RNA, using total serum IgE as template by circular rna circEPSTI1 (hsa_circRNA_000479) specific reverse transcriptases into cDNA, again real-time quantitative PCR amplification is carried out with Specific PCR primers, reference gene is used as using β actin, circEPSTI1 relative quantification Δ CT values are obtained, the circEPSTI1 expression positives are prompted as Δ CT≤5.52.The present invention expresses situation by circEPSTI1 in the doubtful three cloudy breast cancer tissue's sample of detection of relative quantification, for the application in three cloudy breast cancer patients prognosis predictions.
Description
Technical field
The present invention relates to breast cancer diagnosis technical field, specifically discloses a kind of small molecule non-coding circular rna
CircEPSTI1 and its application in three cloudy breast cancer.
Background technology
Breast cancer is whole world incidence of disease highest female malignant.China is as populous nation and unique aging
Social pattern, breast cancer incidence growth rate are higher than other countries, and situation allows of no optimist.Breast cancer is so far still without being filled
Divide effective control, the higher death rate is still presented in breast cancer, and main cause is its recurrence and transfer, and breast cancer once occurs
Transfer, average viability probably only have 4-5.Three cloudy breast cancer refer to ERs (ER), progesterone receptor (PR) and people
The negative breast cancer of skins growth factor receptors -2 (HER-2) expression, account for the 15%-20% of whole breast cancer colony.Three
Cloudy breast cancer Clinical symptoms is not showed only as that aggressive strong, histological grade is higher, poor prognosis, the visceral metastases that easily occur, and lacks
The corresponding target spot of weary endocrine therapy and molecular targeted therapy for HER-2, the selection for the treatment of means is relative to other molecules
Hypotype breast cancer is restricted.
The content of the invention
In order to solve the above problems, it is an object of the invention to provide it is a kind of can in triple negative breast cancer high expression ring
Shape RNA circEPSTI1.
The second object of the present invention there is provided how by detecting RNA circEPSTI1 in tissue, to three negative breasts
The specific method that cancer is diagnosed.
The third object of the present invention there is provided the kit as made from RNA circEPSTI1, the kit cost performance
Application high, easy to spread.
The fourth object of the present invention there is provided the RNA circEPSTI1 and can be applied to three cloudy breast cancer patients prognosis
Prediction.
The present invention is achieved through the following technical solutions：
A kind of circular rna circEPSTI1, the circEPSTI1 are small molecule non-coding ring-type, from EPSTI1
The exon region of gene, its sequence is as shown in SEQ No: 1.
RNA circEPSTI1 belong to one kind of endogenous non-coding RNA, and its structure is single-stranded for covalent annular closed
RNA.With features such as Stability Analysis of Structures, abundance height and tissue specific expressions.It sends out as competitive endogenous RNA (ceRNA)
Wave the effect of gene expression regulation.Circular rna utilizes its microRNA (miRNA) response element combination miRNA, to block
The inhibitory action that miRNA is expressed its target, so as to regulate and control other related RNA expression.Circular rna of the present invention
CircEPSTI1 has main biological function as protein coding gene, be the new judgement prognostic marker of tumour and
The pretty good instrument of targeted therapy, the high expression especially in three cloudy breast cancer, defeats triple negative breast cancer to provide newly for the mankind
Strategy.
The application of the RNA circEPSTI1, by the circEPSTI1 three cloudy breast cancer of diagnosis, specific method is such as
Under：
(1) tissue RNA extracting
A takes tissue specimen to add liquid nitrogen grinding in mortar, adds Trizol and chloroform after being ground into homogenate, concussion is uniform
Place on ice afterwards；
B by step A gained mixed liquor low-temperature centrifugation, take upper strata aqueous phase add precooling isopropanol mix, refrigerator stand
Low-temperature centrifugation afterwards；Supernatant is abandoned, the water-reducible ethanol of 75%DEPC is added and mixes, low-temperature centrifugation；Supernatant is abandoned, is obtained after drying at room temperature
RNA；
C is first dissolved toward addition DEPC water in gained RNA in step B, then RNA concentration and quality are measured with spectrophotometric,
OD260/280 ratios are between 1.6-1.8 and carry out EB gel electrophoresis detection RNA mass, obtain the tissue RNA, -70 DEG C of guarantors
Deposit；
(2) specific reverse transcriptase
Reverse transcription is carried out to circEPSTI1 using transcript reagent box and reverse transcription specific primer, reverse transcription is anti-carrying out
The various reagents in addition to reverse transcriptase must be mixed into reverse transcription mixed liquor before answering, the pipe of installed reagents is flicked with finger,
Reverse transcription mixed liquor is inhaled with pipettor and beaten several times, it is impossible to uses oscillator.
It is preferred that described use transcript reagent box and reverse transcription specific primer to circEPSTI1 progress reverse transcriptions, point
Not Wei Pu Luomaige Promega companies Reverse Transcriptase kit A3500 and Shanghai Ji Ma Bioisystech Co., Ltd production it is inverse
Transcribe specific primer QPM010.
The system of 20 μ L reverse transcription reactions is as follows：
Reverse transcription process：16 DEG C 30 minutes, 42 DEG C 30 minutes, 85 DEG C 10 minutes.
(3) circEPSTI1 specificity real-time quantitative PCR
CircEPSTI1 reverse transcription products are first diluted to 2 times respectively, then mixed；
In circEPSTI1 specificity real-time quantitative PCRs, 20 μ L reaction systems are as follows：
CircEPSTI1 real-time quantitative PCR response procedures：95 DEG C 3 minutes, 40 circulation, 95 DEG C 12 seconds, 62 DEG C 35 seconds, make
Real-time quantitative PCR amplification is carried out with SYBR-Green dyestuffs.
The circEPSTI1 Specific PCR primers are：
Forward primer is 5 '-AAGCTGAAGAAGCTGAACTC-3 '
Reverse primer is 5 '-GTGTATGCACTTGTGTATTGC-3 '；
β-actin the Specific PCR primers are：
Forward primer is 5 '-CATGTACGTTGCTATCCAGGC-3 '
Reverse primer is 5 '-CTCCTTAATGTCACGCACGAT-3 '.
(4) measure of △ CT indexs
As △ CT≤5.52, circEPSTI1 expression is positive, you can diagnoses this and is organized as three cloudy breast cancer tissues；It is described
△ CT are the difference of circEPSTI1 and β-actin mean CT-number.
The application of the RNA circEPSTI1, the RNA circEPSTI1 are applied to three cloudy breast cancer detection reagents
Box.Detection kit includes：
Isopropanol, chloroform, Trizol, DEPC water or without enzyme water, distilled water, 5 × RT Buffer, 25mM chlorinations
Magnesium, 10mM triphosphoric acid bases deoxynucleotide, 5U/ μ l RAN enzyme inhibitors, 200U/ μ l MMLV reverse transcriptases or 25U/ μ l
AMV enzymes, 2 × real-time quantitative PCR buffer solution, 5U/ μ l Taq polymerases, 5 μM of circEPSTI1 Specific PCR primers, 5 μM of β-
Actin Specific PCR primers, 10 μM of circular rna reverse transcriptase primers, 10 μM of β-actin specific reverse transcriptase primers.
RNA circEPSTI1 application, it is pre- that the RNA circEPSTI1 are applied to three cloudy breast cancer patients prognosis
Survey.
CircEPSTI1 is hsa_circRNA_000479 (chr13 in the present invention：43528083-43544806), it comes
Come from EPSTI1 genes (13q14.11) exon region (Fig. 1).And No. 13 chromosome where it carry it is some with it is newborn
The Cancer-Related gene of gland, including BRCA2 and RB1 genes etc..Real-Time Fluorescent Quantitative PCR Technique is current detection tissue circular rna
Express one of detection method of authority.Applicant detects 57 triple negative breast cancers by Real-Time Fluorescent Quantitative PCR Technique first
Expression in tissue and its pairing normal galactophore tissue, as a result finds, circEPSTI1 is in triple negative breast cancer tissue
It is middle that high expression (Fig. 2) is presented.CircEPSTI1 is high in triple negative breast cancer tissue to express statistically significant (P<0.01).For
Preferably judge the expression in circEPSTI1 triple negative breast cancer tissues, further analyze circEPSTI1 for three
The influence of cloudy patient with breast cancer's prognosis, the applicant have detected 240 three cloudy breast cancer again by way of in situ hybridization
CircEPSTI1 expression (Fig. 3) in patient tumors.
Follow-up Data shows there are 24 death in 83 triple negative breast cancer patients of the high expression of circEPSTI1, and
Only have 14 death in 157 triple negative breast cancer patients of circEPSTI1 low expressions；Credit is counted through Kaplan-Meier
The triple negative breast cancer patient overall survival for analysing the high expression of circEPSTI1 is less than the patient of circEPSTI1 low expressions, and difference is equal
Statistically significant (P<0.01) (Fig. 4), circEPSTI1 high expression in triple negative breast cancer is prompted, is the negative breast of prediction three
The molecular marker of gland cancer prognosis.The present invention provides strong technical support and molecule life for triple negative breast cancer prognosis prediction
Thing basis, has far-reaching clinical meaning and generalization.
In previous research work, the applicant utilizes circular rna high flux chip (Arraystar HumancircRNA
Array V2) in 3 pair of three cloudy breast cancer tissue circular rna related to screening triple negative breast cancer in the Carcinoma side normal tissue matched
Differential expression is composed.It is more than 2 times according to fold differences, P<0.05 standard, it is found that triple negative breast cancer correlation circular rna there are 92,
Wherein up-regulation circular rna has 68, and lowering circular rna has 24.The circular rna of subsequent 5 up-regulations of picking is negative at 51 pair three
Breast cancer tissue in the Carcinoma side normal tissue of pairing with carrying out qRT-PCR checkings.As a result show, RNA hsa_circRNA_
000479 (crEPSTI1) is significantly higher than corresponding cancer beside organism in three cloudy expression in breast highests.With chip results
Unanimously.
By further fabric study, inventor has found：High table is presented in circEPSTI1 in triple negative breast cancer tissue
Up to (referring to accompanying drawing 2,3).The triple negative breast cancer patient overall survival of the high expression of circEPSTI1 is less than the low tables of circEPSTI1
The patient reached (referring to accompanying drawing 4).This prompting circEPSTI1 is the molecular marker for predicting triple negative breast cancer prognosis.
The present invention's confirms circEPSTI1 high expression in triple negative breast cancer by studying, and can be used for three negative breasts
A kind of application in carninomatosis people's prognosis prediction, applied to field of medicaments, there is provided the negative breast of the three of cost-effective, easy to spread application
Gland cancer diagnostic kit.
Figure of description
Fig. 1 circEPSTI1 schematic diagrames
Fig. 2 real-time fluorescence quantitative PCRs are surveyed expression of the circEPSTI1 in triple negative breast cancer tissue and changed
Fig. 3 in situ hybridizations detect expression of the circEPSTI1 in triple negative breast cancer tissue and changed
Fig. 4 circEPSTI1 and the relation of triple negative breast cancer patient's prognosis
Specific embodiment
With reference to embodiment, the present invention is described in further detail, is managed to help those skilled in the art
The solution present invention.
First, triple negative breast cancer organizational diagnosis kit forms (50 secondary response)
Isopropanol 100ml, chloroform 100ml, Trizol 50ml, DEPC water or without enzyme water 10ml, distilled water 10ml, 5
× RT Buffer 1ml, 25mM magnesium chloride 1ml, 10mM triphosphoric acid base deoxynucleotide 1ml, 5U/ μ l RAN enzyme inhibitors
500 50 μ l or 25U/ μ lAMV enzymes of μ l, 200U/ μ l MMLV reverse transcriptases 50 μ l, 2 × real-time quantitative PCR buffer solution 2ml, 5U/ μ l
The μ l of Taq polymerase 50,
5 μM of μ l of circEPSTI1 Specific PCR primers 50,
Its forward primer is 5 '-AAGCTGAAGAAGCTGAACTC 3 ',
Its reverse primer is 5 '-GTGTATGCACTTGTGTATTGC-3 '.
5 μM of μ l of β-actin Specific PCR primers 30
Its forward primer is 5 '-CATGTACGTTGCTATCCAGGC-3 '
Its reverse primer is 5 '-CTCCTTAATGTCACGCACGAT-3 '
10 μM of μ l of circular rna reverse transcriptase primer 20 (Shanghai Ji Ma Bioisystech Co., Ltd, QPM010).
10 μM of μ l of β-actin specific reverse transcriptases primer 20 (Shanghai Ji Ma Bioisystech Co., Ltd, QPM010).
2nd, in tissue samples circEPSTI1 detection
1st, RNA extracting is organized
Tissue specimen is taken with adding liquid nitrogen grinding sample in mortar；0.6ml Trizol mortar samples are added in mortar,
Medication spoon is added in tube pipes after being ground into homogenate；0.4ml Trizol are added in tube pipes；Chlorination imitates 200 μ l/mlTrizol
In Tube, 15-30s is shaken with hand, places 5min on ice, 4 DEG C of 12000g centrifuge 15min；Carefully upper strata aqueous phase is taken to enter newly
In tube, the isopropanol 0.5ml/mlTrizol for adding precooling is mixed, and -20 DEG C of refrigerators stand 20min, 4 DEG C of 12000g centrifugations
10min；Supernatant is abandoned, the water-reducible ethanol 1-2ml of 75%DEPC is added and mixes, 4 DEG C of 7500g centrifuge 5min, abandon supernatant, room as far as possible
Temperature dries 5-10min, adds DEPC water 10-20 μ l dissolvings RNA.Spectrophotometric measures RNA concentration and quality, OD260/280
Ratio is between 1.6-1.8 and carries out EB gel electrophoresis detection RNA mass, and -70 DEG C preserve.
2nd, specific reverse transcriptase：Use the Reverse Transcriptase kit (A3500) and Shanghai Ji agate of Pu Luomaige Promega companies
The reverse transcription specific primer (QPM010) of Bioisystech Co., Ltd's production carries out reverse transcription to circEPSTI1.20 μ L are reversed
The system for recording reaction is as follows：
Various reagents in addition to reverse transcriptase must be mixed into reverse transcription mixed liquor before reverse transcription reaction is carried out, used
Finger flicks the pipe of installed reagents, and reverse transcription mixed liquor is inhaled with pipettor and beaten several times, it is impossible to uses oscillator.
Reverse transcription process：16 DEG C 30 minutes, 42 DEG C 30 minutes, 85 DEG C 10 minutes.
3rd, circEPSTI1 specificity real-time quantitative PCR：CircEPSTI1 reverse transcription products are first diluted to 2 times respectively,
Then mix.20 μ L reaction systems are as follows：
CircEPSTI1 real-time quantitative PCR response procedures：95 DEG C 3 minutes, 40 circulation, 95 DEG C 12 seconds, 62 DEG C 35 seconds.
Real-time quantitative PCR amplification is carried out using SYBR-Green dyestuffs.
CircEPSTI1 PCR specific primers are：
Its forward primer is 5 '-AAGCTGAAGAAGCTGAACTC 3 ',
Its reverse primer is 5 '-GTGTATGCACTTGTGTATTGC-3 '.
β-actin are used as reference gene, and its PCR primer sequence is：
Its forward primer is 5 '-CATGTACGTTGCTATCCAGGC-3 '
Its reverse primer is 5 '-CTCCTTAATGTCACGCACGAT-3 '
(4)△CTThe measure of index：△CTRefer in same sample, circEPSTI1 to be checked and internal reference β-actin Average Ct values
Difference.That is circEPSTI1 △ CT=circEPSTI1MeanCT-control MeanCT, in of the invention, △ CTFor
CircEPSTI1 and β-actin average CTThe difference of value, relative quantification Δ CT values are obtained, and judged, as a result shown,
In 57 triple negative breast cancer tissues of detection, as △ CTWhen≤5.52, the circEPSTI1 expression positive has 39, and positive rate is
68.42%.
3rd, the Prognostic significance of circEPSTI1 and triple negative breast cancer patient
With hybridization in situ technique, applicant have detected the expression of circEPSTI1 in 240 triple negative breast cancer tissues
Situation, prognosis follow-up then is carried out to the triple negative breast cancer patient of each detection, as a result show the high expression of circEPSTI1
Triple negative breast cancer patient overall survival be less than circEPSTI1 low expressions patient (referring to accompanying drawing 4).This prompting
CircEPSTI1 is the molecular marker for predicting triple negative breast cancer prognosis.
Research shows above, detects the expression of circEPSTI1 in triple negative breast cancer tissue, has good stability.
CircEPSTI1 can be cloudy as the specificity molecular marker of triple negative breast cancer patient's prognosis prediction, circEPSTI1 and three
Property Prognosis in Breast Cancer is significantly correlated, the application that can be used in triple negative breast cancer patient's prognosis prediction.
Above-described embodiment, simply presently preferred embodiments of the present invention, is not used for limiting the scope of the present invention, therefore all with this
The equivalent change or modification that feature and principle described in invention claim are done, all should be included in scope of the invention as claimed
Within.
Sequence number
Denomination of invention：A kind of RNAcircEPSTI1 and its application applicant in three cloudy breast cancer：Zhongshan University's tumour
Centre of prevention and cure
RNA circEPSTI1 sequence numbers
CACAAGTGCATACACCTTGATAGCACCAAATATAAACCGGAGAAATGAGATACAAAGAATTGCGGAGCA
GGAGCTGGCCAACCTGGAGAAGTGGAAGGAGCAGAACAGAGCTAAACCGGTTCACCTGGTGCCCAGACGGCTAGGTG
GAAGCCAGTCAGAAACTGAAGTCAGACAGAAACAACAACTCCAGCTGATGCAATCTAAATACAAGCAAAAGCTAAAA
AGAGAAGAATCTGTAAGAATCAAGAAGGAAGCTGAAGAAGCTGAACTCCAAAAAATGAAGGCAATTCAGAGAGAGAA
GAGCAATAAACTGGAGGAGAAAAAAAGACTTCAAGAAAACCTTAGAAGAGAAGCATTTAGAGAGCATCAGCAATA
Sequence number
Denomination of invention：A kind of RNA circEPSTI1 and its application in three cloudy breast cancer
Applicant：Zhongshan Univ. Cancer Cure Center
RNA circEPSTI1 sequence numbers
CACAAGTGCATACACCTTGATAGCACCAAATATAAACCGGAGAAATGAGATACAAAGAAT
TGCGGAGCAGGAGCTGGCCAACCTGGAGAAGTGGAAGGAGCAGAACAGAGCTAAACCGGT
TCACCTGGTGCCCAGACGGCTAGGTGGAAGCCAGTCAGAAACTGAAGTCAGACAGAAACA
ACAACTCCAGCTGATGCAATCTAAATACAAGCAAAAGCTAAAAAGAGAAGAATCTGTAAG
AATCAAGAAGGAAGCTGAAGAAGCTGAACTCCAAAAAATGAAGGCAATTCAGAGAGAGAA
GAGCAATAAACTGGAGGAGAAAAAAAGACTTCAAGAAAACCTTAGAAGAGAAGCATTTAG
AGAGCATCAGCAATA
Claims (10)
- A kind of 1. circular rna circEPSTI1, it is characterised in that；The circEPSTI1 is small molecule non-coding ring-type, source In the exon region of EPSTI1 genes, its sequence is as shown in SEQ No: 1.
- 2. the application of RNA circEPSTI1 described in claim 1, it is characterised in that；The circEPSTI1 is cloudy with diagnosis three Breast cancer, specific method are as follows：(1) tissue RNA extractingA takes tissue specimen to add liquid nitrogen grinding in mortar, adds Trizol and chloroform after being ground into homogenate, ice after shaking uniformly Upper placement；B by step A gained mixed liquor low-temperature centrifugation, take upper strata aqueous phase add precooling isopropanol mix, refrigerator stand after it is low Temperature centrifugation；Supernatant is abandoned, the water-reducible ethanol of 75%DEPC is added and mixes, low-temperature centrifugation；Supernatant is abandoned, RNA is obtained after drying at room temperature；C is first dissolved toward addition DEPC water in gained RNA in step B, then RNA concentration and quality are measured with spectrophotometric, OD260/280 ratios are between 1.6-1.8 and carry out EB gel electrophoresis detection RNA mass, obtain the tissue RNA, -70 DEG C of guarantors Deposit；(2) specific reverse transcriptaseReverse transcription is carried out to circEPSTI1 using transcript reagent box and reverse transcription specific primer, carry out reverse transcription reaction it It is preceding the various reagents in addition to reverse transcriptase to be mixed into reverse transcription mixed liquor, the pipe of installed reagents is flicked with finger, will be inverse Transcription mixed liquor is inhaled with pipettor to be beaten several times, it is impossible to uses oscillator；(3) circEPSTI1 specificity real-time quantitative PCRCircEPSTI1 reverse transcription products are first diluted to 2 times respectively, then mixed；(4) measure of △ CT indexsAs △ CT≤5.52, circEPSTI1 expression is positive, you can diagnoses this and is organized as three cloudy breast cancer tissues；The △ CT For circEPSTI1 and β-actin mean CT-number difference.
- 3. RNA circEPSTI1 as claimed in claim 2 application, it is characterised in that：In (2) specific reverse transcriptase, 20 The system of μ L reverse transcription reactions is as follows：Reverse transcription process：16 DEG C 30 minutes, 42 DEG C 30 minutes, 85 DEG C 10 minutes.
- 4. RNA circEPSTI1 as claimed in claim 2 application, it is characterised in that：(3) the circEPSTI1 specificity In real-time quantitative PCR, 20 μ L reaction systems are as follows：CircEPSTI1 real-time quantitative PCR response procedures：95 DEG C 3 minutes, 40 circulation, 95 DEG C 12 seconds, 62 DEG C 35 seconds, use SYBR-Green dyestuffs carry out real-time quantitative PCR amplification.
- 5. the application of RNA circEPSTI1 described in claim 1, it is characterised in that；The RNA circEPSTI1 are applied In three cloudy breast cancer detection kits.
- 6. RNA circEPSTI1 as claimed in claim 5 application, it is characterised in that：The three cloudy breast cancer detection kit Including：Trizol, chloroform, isopropanol, water, RT Buffer, magnesium chloride, triphosphoric acid base deoxynucleotide, RNase Inhibitor, enzyme, circEPSTI1 Specific PCR primers, β-actin Specific PCR primers, real-time quantitative PCR buffer solution, Taq Polymerase, circular rna reverse transcriptase primer, β-actin specific reverse transcriptase primers；The water be without the one or more combination thing in enzyme water, DEPC water or distilled water, the enzyme be MMLV reverse transcriptases or AMV enzymes.
- 7. RNA circEPSTI1 as claimed in claim 6 application, it is characterised in that the three cloudy secondary response of breast cancer 50 Detection kit includes：Isopropanol 100ml, chloroform 100ml, Trizol 50ml, DEPC water or without enzyme water 10ml, distilled water 10ml, 5 × it is inverse Transcription buffer 1ml, 25mM magnesium chloride 1ml, 10mM triphosphoric acid base deoxynucleotide 1ml, the μ of 5U/ μ l RAN enzyme inhibitors 500 L, μ l of 50 μ l or 25U/ μ l AMV enzymes of 200U/ μ l MMLV reverse transcriptases 50,0.2 × real-time quantitative PCR buffer solution 2ml, 5U/ μ l μ l of Taq polymerase 50,5 μM of μ l of circEPSTI1 Specific PCR primers 50,5 μM of μ l of β-actin Specific PCR primers 30,10 μ μ l of M circular rnas reverse transcriptase primer 20,10 μM of μ l of β-actin specific reverse transcriptases primer 20.
- 8. the RNA circEPSTI1 application as described in any one in claim 2,6 or 7, it is characterised in that：It is described CircEPSTI1 Specific PCR primers are：Forward primer is 5 '-AAGCTGAAGAAGCTGAACTC-3 'Reverse primer is 5 '-GTGTATGCACTTGTGTATTGC-3 '；β-actin the Specific PCR primers are：Forward primer is 5 '-CATGTACGTTGCTATCCAGGC-3 'Reverse primer is 5 '-CTCCTTAATGTCACGCACGAT-3 '.
- 9. the RNA circEPSTI1 application as described in any one in claim 2,6 or 7, it is characterised in that：It is described right CircEPSTI1 carries out reverse transcription and uses transcript reagent box and reverse transcription specific primer, respectively Pu Luomaige Promega public affairs The Reverse Transcriptase kit A3500 of the department and reverse transcription specific primer QPM010 of Shanghai Ji Ma Bioisystech Co., Ltd production.
- 10. the application of RNA circEPSTI1 described in claim 1, it is characterised in that；Should by the RNA circEPSTI1 For three cloudy breast cancer patients prognosis predictions.
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Cited By (6)
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| CN109097477A (en) * | 2018-09-10 | 2018-12-28 | 山东大学齐鲁医院 | It is a kind of for the circRNA marker of breast cancer diagnosis and its application |
| CN109280706A (en) * | 2018-12-11 | 2019-01-29 | 宁夏医科大学总医院 | A kind of circular rna hsa-circ-0102618 and its specificity amplification primer and application |
| CN109295059A (en) * | 2018-09-13 | 2019-02-01 | 中山大学肿瘤防治中心（中山大学附属肿瘤医院、中山大学肿瘤研究所） | A kind of single stranded RNA of covalent annular closed and its application |
| CN109371025A (en) * | 2018-12-11 | 2019-02-22 | 宁夏医科大学总医院 | A kind of circular rna hsa-circ-0044508 and its specificity amplification primer and application |
| CN109402122A (en) * | 2018-12-11 | 2019-03-01 | 宁夏医科大学总医院 | A kind of circular rna hsa-circ-0044551 and its specificity amplification primer and application |
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| CN109295059B (en) * | 2018-09-13 | 2022-06-14 | 中山大学肿瘤防治中心（中山大学附属肿瘤医院、中山大学肿瘤研究所） | Application of covalent circular closed single-stranded RNA in triple-negative breast cancer kit |
| CN109280706A (en) * | 2018-12-11 | 2019-01-29 | 宁夏医科大学总医院 | A kind of circular rna hsa-circ-0102618 and its specificity amplification primer and application |
| CN109371025A (en) * | 2018-12-11 | 2019-02-22 | 宁夏医科大学总医院 | A kind of circular rna hsa-circ-0044508 and its specificity amplification primer and application |
| CN109402122A (en) * | 2018-12-11 | 2019-03-01 | 宁夏医科大学总医院 | A kind of circular rna hsa-circ-0044551 and its specificity amplification primer and application |
| CN110923321A (en) * | 2019-12-26 | 2020-03-27 | 广东省人民医院(广东省医学科学院) | Annular RNA detection kit for predicting neoadjuvant chemotherapy reactivity of triple-negative breast cancer |
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