CN109457033A - Marker for colon cancer screening - Google Patents
Marker for colon cancer screening Download PDFInfo
- Publication number
- CN109457033A CN109457033A CN201811641665.5A CN201811641665A CN109457033A CN 109457033 A CN109457033 A CN 109457033A CN 201811641665 A CN201811641665 A CN 201811641665A CN 109457033 A CN109457033 A CN 109457033A
- Authority
- CN
- China
- Prior art keywords
- circ
- hsa
- colon cancer
- screening
- seq
- 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.)
- Granted
Links
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
-
- 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
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/6851—Quantitative amplification
-
- 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
- C12Q2600/118—Prognosis of disease development
-
- 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
- C12Q2600/166—Oligonucleotides used as internal standards, controls or normalisation probes
-
- 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
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Immunology (AREA)
- Analytical Chemistry (AREA)
- Genetics & Genomics (AREA)
- Physics & Mathematics (AREA)
- Pathology (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- General Health & Medical Sciences (AREA)
- General Engineering & Computer Science (AREA)
- Biophysics (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Oncology (AREA)
- Hospice & Palliative Care (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention discloses the markers for colon cancer screening, belong to molecular diagnosis field.The present invention develops new molecular marker hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_0086569 for colon cancer screening diagnosis, the diagnostic value of the single circRNA of marker is higher, ROC curve has good sensitivity and specificity analysis shows that it can preferably distinguish screening colorectal cancer patients；Diagnostic of the molecular marked compound express spectra that 3 circRNA join together to form in terms of diagnosing colon than single circRNA molecule is higher, is expected to become the important indicator for being applied to colorectal cancer clinical diagnosis and treatment assessment.
Description
Technical field
The invention belongs to biomedical and technical field of molecular biology, and in particular to the Clinical screening field of colon cancer,
More specifically, the present invention relates to the newfound marker for colon cancer screening and the molecular agents for colon cancer screening
Box.
Background technique
It should be noted that this part content is only the reference content and background of invention of present invention design part, not
Necessarily constitute the disclosure relative to the prior art of the invention.
Colon cancer is one of main malignant tumour of Digestive, in recent years, with the improvement of people's living standard in our country, is drunk
The aggravation of change, the aging of population of structure is eaten, disease incidence of the colon cancer in China is showed an increasing trend year by year, seriously threatened
People's health.Though the cause of disease of colon cancer is not fully apparent from, some factors such as environment, dietary structure, living habit (suction of drinking
Cigarette) etc. may induce colon cancer, in addition, familial polyposis, ulcerative colitis and familial neoplasms heredity medication history etc. are also
The high risk factor of colon cancer occurs.
The existence and prognosis of colorectal cancer patients depend on the detection time of tumour, and colon cancer early stage, itself was without specific disease
Shape, patient's physical examination, consciousness of going to a doctor in time is not high in addition, and leading to segmental colonic cancer has been advanced stage when finding, comes to treatment zone very big
Difficulty, prognosis are relatively poor.Data statistics be more than when 57% patient makes a definite diagnosis cancer cell shifted, and early stage colorectal cancer patients 5
Year survival rate can reach 90%, but advanced stage and 5 years overall survivals of metastatic patient be only 15%.Suffer to improve colon cancer
The survival rate of person clinically sets about the diagnostic method and diagnostic criteria of studying and defining early stage colon cancer, at present diagnosis of colon cancer master
It to be checked dependent on colonoscopy, iconography and fecal occult blood etc., but still lack corresponding convenience, sensibility and specificity.
With the fast development of Protocols in Molecular Biology and deepening continuously for colorectal cancer Mechanism Study, more and more phases
Biomarker is closed constantly to be found and reported.From the point of view of the progress of non-coding RNA, some miRNA, lncRNA,
CircRNA has the application prospect as disease biomarkers.Circular rna (CircularRNA, circRNA) is a kind of wide
It is general to be present in the intracorporal single-stranded non-coding RNA of biology, it is found in the RNA virus of the 1970s earliest, with traditional line
Property RNA it is different, circRNA molecule is in closed circular structure, is not influenced by RNA excision enzyme, express it is more stable, it is not degradable；?
Functionally, circRNA molecule is rich in microRNA binding site, plays the role of miRNA sponge in cell, and then release
MiRNA increases the expression of target gene to the inhibiting effect of its target gene；By interacting with the miRNA of disease association,
CircRNA plays important regulating and controlling effect in disease.
It is related with colon cancer that a variety of circRNA are had also discovered at present.Weng W etc. analyzes two groups of independent colorectal cancers
CiRS-7 expresses the influence to prognosis in patient, it is found that high ciRS-7 expression is related with overall low survival rate；Dou Y etc. has studied
The relationship of KRAS mutation and colon cancer cell finds circRTN4 in the excretion body of colon cancer cell DLD-1 in high expression
Level, and it is then in downward state in the cell；In addition to this, circ-ITCH, hsa_circ_0014717, circHIPK3,
circCCDC66、hsa_circ_0001649、circ-KLDHC10、hsa_circ_0000567、hsa_circ_000984、
circRNA0003906、has_circ_0020397、circCCDC66、hsa_circ_0000069、hsa_circ_001569、
hsa_circ_001988、circ-BANP、hsa_circRNA_103809、hsa_circRNA_104700、hsa_circ_
The 0126897 equal occurrence and development for being also found participation colon cancer invade transfer process, as colon cancer prognostic marker or influence
Colon cancer Concurrent Chemoradiotherapy Sensitivity etc..But detected for colon cancer screening, it is also necessary to new effective marker is further explored,
And explore its specificity and accuracy.
Summary of the invention
Based on above content, the present invention obtains the circRNA marker for colon cancer screening after study, and is based on
This develops the Related product for colon cancer screening.
Specifically, the invention discloses following technical schemes:
Firstly, the invention discloses in hsa_circ_0036847, hsa_circ_0064628, hsa_circ_0086569
One or more combination as marker preparation for colon cancer screening product in application.
Secondly, the product can be by detecting patient's colon the invention discloses a kind of product for colon cancer screening
Hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_0086569 expression in tissue carrys out screening colon
Cancer, testing result show that hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_0086569 expression are more normal
The significant up-regulation of tissue.
Further, the invention discloses the fluorescence quantification PCR primers of the circRNA marker for colon cancer screening
Group, the primer sets are respectively used for amplifying hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_0086569.
In addition, passing through hsa_circ_0036847, hsa_circ_ the invention also discloses the method for colon cancer screening
The relative expression quantity of 0064628 or hsa_circ_0086569 judges to suffer from the risk of colon cancer.
Further, present invention provides hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_
Application of one or more of 0086569 combination in the drug of preparation colon cancer prognosis or treatment.
Based on technical solution of the present invention, the present invention at least achieves following technical effect:
(1) present invention develop for colon cancer screening diagnosis new molecular marker hsa_circ_0036847,
Hsa_circ_0064628 or hsa_circ_0086569 provides new reference for the screening of colon cancer；
(2) marker hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_ of the present invention
In 0086569, the diagnostic value of single circRNA is higher, and ROC curve is analysis shows that it can preferably distinguish screening colon
Cancer patient, the area under the curve of three are respectively 0.906,0.853,0.961, all have good sensitivity and specificity；Into
One step, the molecular marked compound express spectra that 3 circRNA join together to form is in terms of diagnosing colon than single circRNA
The diagnostic of molecule is higher, is expected to become the important indicator for being applied to colorectal cancer clinical diagnosis and treatment assessment.
Detailed description of the invention
Fig. 1 is functional annotation of the hsa_circ_0036847 in parental gene
Fig. 2 is functional annotation of the hsa_circ_0064628 in parental gene
Fig. 3 is functional annotation of the hsa_circ_0086569 in parental gene
Fig. 4 is relative expression quantity statistics of the hsa_circ_0036847 in normal tissue and colon cancer tissue
Fig. 5 is relative expression quantity statistics of the hsa_circ_0064628 in normal tissue and colon cancer tissue
Fig. 6 is relative expression quantity statistics of the hsa_circ_0086569 in normal tissue and colon cancer tissue
Fig. 7 is the diagnostic ROC curve of hsa_circ_0036847
Fig. 8 is the diagnostic ROC curve of hsa_circ_0064628
Fig. 9 is the diagnostic ROC curve of hsa_circ_0086569
Specific embodiment
It is noted that described further below be all exemplary, it is intended to provide further illustrate to the disclosure.Unless another
It indicates, all technical and scientific terms used herein has usual with disclosure person of an ordinary skill in the technical field
The identical meanings of understanding.
It should be noted that term used herein above is merely to describe specific embodiment, and be not intended to restricted root
According to the illustrative embodiments of the disclosure.Additionally, it should be understood that when in the present specification use term "comprising" and/
Or when " comprising ", existing characteristics, step, operation and/or their combination are indicated.
In one disclosed by the invention or some typical embodiments, hsa_circ_0036847, hsa_ are disclosed
One or more of circ_0064628, hsa_circ_0086569 combination are used for colon cancer screening in preparation as marker
Product in application.
The sequence of hsa_circ_0036847 (spliced seq) as shown in SEQ ID NO:1, the position in genome
For chr15:91083270-91157720, the functional annotation in parental gene is as shown in Figure 1；
For the sequence of hsa_circ_0064628 as shown in SEQ ID NO:2, the position in genome is chr3:
28373179-28382113, the functional annotation in parental gene are as shown in Figure 2；
For the sequence of hsa_circ_0086569 as shown in SEQ ID NO:3, the position in genome is chr9:
20923658-20926416, the functional annotation in parental gene are as shown in Figure 3.
In specific embodiment, the product for colon cancer screening includes the hsa_ detected in patient's colonic tissue
The reagent or drug of circ_0036847, hsa_circ_0064628 or hsa_circ_0086569 expression.
In some typical embodiments, the product for colon cancer screening includes: by real-time quantitative PCR, original position
Hybridization, genetic chip or gene sequencing detect hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_
0086569 expression is with the product of diagnosing colon.
In one disclosed by the invention or some typical embodiments, a kind of production for colon cancer screening is disclosed
Product, the product can pass through hsa_circ_0036847, hsa_circ_0064628 or hsa_ in detection patient's colonic tissue
Circ_0086569 expression carrys out screening colon cancer, and testing result shows hsa_circ_0036847, hsa_circ_
0064628 or hsa_circ_0086569 expression is significantly raised compared with normal tissue.
In colorectal cancer patients, the person that has polyp of colon, colon cancer incidence is usually 5 times without polyp of colon person,
In a specific embodiment of the invention, patient's colonic tissue can be selected from polyp of colon or its hetero-organization (such as colon epithelial cell
Deng).
In the specific embodiment of the invention, the fluorescent quantitation of the circRNA marker for colon cancer screening is disclosed
PCR primer group, the primer sets are respectively used for amplifying hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_
0086569。
In specific embodiment, the primer sets sequence for expanding hsa_circ_0036847 includes SEQ ID NO:4
With SEQ ID NO:5；Primer sets sequence for expanding hsa_circ_0064628 includes SEQ ID NO:6 and SEQ ID NO:
7；Primer sets sequence for expanding hsa_circ_0086569 includes SEQ ID NO:8 and SEQ ID NO:9.
In a specific embodiment of the invention, the kit of the circRNA marker for colon cancer screening is disclosed,
Including above-mentioned fluorescence quantification PCR primer group.
In specific embodiment, the primer sets of amplification reference gene can also be specifically included, such as reference gene can be with
It is GAPDH；Primer sets sequence for expanding GAPDH includes SEQ ID NO:10 and SEQ ID NO:11.
In one disclosed by the invention or some typical embodiments, the method for colon cancer screening is disclosed, is passed through
The relative expression quantity of hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_0086569 judge to suffer from colon
The risk of cancer.
In the specific embodiment of the invention, pass through hsa_circ_0036847, hsa_ in detection patient's colonic tissue
Circ_0064628 or hsa_circ_0086569 expression carrys out screening colon cancer, and testing result shows hsa_circ_
0036847, hsa_circ_0064628 or hsa_circ_0086569 expression is significantly raised compared with normal tissue, then it represents that suffers from knot
Intestinal cancer.
In one disclosed by the invention or some typical embodiments, usual colon cancer tissue circRNA's is opposite
Expression difference is also related with the prognosis of colorectal cancer patients or treatment, thus also disclosed is hsa_circ_0036847,
One or more of hsa_circ_0064628 or hsa_circ_0086569 combination are in preparation colon cancer prognosis or treatment
Application in product.
In some typical embodiments, the product of the colon cancer prognosis or treatment includes: by real-time quantitative PCR, original
Position hybridization, genetic chip or gene sequencing detect hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_
The product of 0086569 expression.
In order to clearly show that technical solution disclosed by the invention, following present invention will in conjunction with specific embodiments in detail
Describe bright disclosed embodiment and technology contents in detail.
Embodiment
The present invention is based on seminar's early periods by colon cancer screening, chooses group by several colorectal cancer patients tissues and cancer
It is good to be screened out from it several groups of new high different expressions for the deep sequencing research for knitting RNA-seq between (normal tissue)
Circular RNA molecule is to make screening verifying analysis.The present invention have chosen hsa_circ_0036847, hsa_circ_0064628 and
Hsa_circ_0086569 is studied by the expression between colorectal cancer patients tissue and normal tissue, in conjunction with statistics credit
Analysis and verifying, with the screening for colon cancer.
Research object and sample
It is 60 total to choose the colorectal cancer patients accepted for medical treatment in May, -2018 Shandong Tumor Hospital's in June, 2017, wherein male
Property 36, women 24, made a definite diagnosis through pathological examination, the judgement of histopathology type is referring to WHO pathological staging；It is obtained in art
Colon cancer tissue and cancer beside organism's sample, and pursue the proper method preservation (RNA is avoided to degrade).
RNA extracting
Cancerous tissue/cancer beside organism is taken, is gone in the centrifuge tube of 1.5ml after being ground using liquid nitrogen method, the TRIzol of 750 μ l
LS Reagent reagent and 20 μ l glacial acetic acid, acutely oscillation tube body extremely mixes manually.
Sample is incubated for 5 minutes in 15 to 30 DEG C after homogenate.
The chloroform of 0.2ml is added in the sample of the TRIzol LS Reagent reagent homogenate of every 750 μ l.
Manually acutely oscillation tube body 15 seconds after, 15 to 30 DEG C incubation 2-3 minutes.
12000 × g is centrifuged 15 minutes at 4 DEG C.Mixing liquid is divided into the red phenol chloroform phase of lower layer after centrifugation, middle layer and
The colourless aqueous phase on upper layer.RNA is all distributed in water phase.
Water phase is transferred in new centrifuge tube.Add the isopropanol (1ml TRIZOL reagent) of 0.5ml, water phase and isopropanol are mixed
It closes to precipitate RNA therein.
It is incubated for 10 minutes for 15 to 30 DEG C after mixing, is centrifuged 10 minutes in 4 DEG C of 12000 × g.
Supernatant is removed, 75% ethyl alcohol of at least 1ml is added in the sample of every 1ml TRIZOL reagent homogenate, cleans RNA
Precipitating.After oscillation, 4 DEG C of 7500 × g are centrifuged 5 minutes.
Removal ethanol solution, air drying RNA precipitate 5-10 minutes.Partly soluble RNA sample A260/280 ratio
It will be less than 1.6.When dissolving RNA, the water that no RNA enzyme is first added is blown and beaten several times repeatedly with rifle, is then incubated for 10 minutes for 55 to 60 DEG C.
The RNA solution of acquisition is stored in -70 DEG C.
CDNA synthesis
Configure in PCR pipe: RNA1-2 μ g, 0.5 μ g/ μ l N6 (random primer) 1 μ l, dNTPs Mix (10mM) 1 μ l adds nothing
The H of RNA enzyme2O to 13.7 μ l of total volume；Mixed liquor 70 DEG C warm bath 5 minutes, then rapid ice bath is placed 2 minutes.
Prepare reverse transcription system, 4 μ l of RT reaction solution 5X First-Strand Buffer sequentially added in centrifuge tube,
1 μ l, RNase Ihibitor of 0.1M DTT, 0.3 μ l, SuperScript III RT, 1 μ l carries out reverse transcription, reaction condition
42 DEG C of 1h, 70 DEG C of 20min, ice bath save cDNA or are placed in -20 DEG C of placements.
Real-time fluorescence quantitative PCR
Realtime PCR reaction system.Shown in system configurations table 1, the primer sets of use are as shown in table 2.
1 Realtime PCR reaction system of table
2 real-time fluorescence quantitative PCR primer of table
PCR plate is placed in progress PCR reaction in Realtime PCR instrument.All indexs are pressed following procedure and are carried out: 95
DEG C, 10min；40 (95 DEG C, 10 seconds of PCR cycle；60 DEG C, 30 seconds).In order to establish the melting curve of PCR product, amplified reaction
After, by 95 DEG C, 10 seconds；60 DEG C, 30 seconds；95 DEG C, program carries out within 15 seconds；GAPDH is as internal reference.Use 2-△△CtMethod calculates.
Statistics and interpretation of result
The present invention carries out statistical analysis using SPSS 17.0 and GraphPad Prism 5 (San Diego, CA).
Student t is examined for analyzing the difference between two groups.Using single factor test variance point between circRNA and clinical pathological factors
Analysis.Using the colon cancer tissue pathological replacement for excision of performing the operation as goldstandard, receiver operating characteristic curve (ROC) is analyzed for determining
The diagnostic value of circ RNA.P < 0.05 is considered to have statistical significance.
3 three circRNA of table and colorectal cancer patients clinical case parameters relationship are analyzed
According to statistical analysis, the expression and cancerous tissue of hsa_circ_0036847 and hsa_circ_0086569 break up
There is certain relationship, 3 circRNA expressions are unrelated with gender, age, Tumor size, Lymph Node Metastasis situation and TNM stage
(indifference is anisotropic between group).
Real-time quantitative PCR data are shown, in institute's collecting sample of the present invention, compare normal tissue, 3 kinds in colon cancer tissue
The obvious up-regulation of circRNA (hsa_circ_0036847, hsa_circ_0064628, hsa_circ_0086569) expression, ginseng
See Fig. 4-Fig. 6 (P < 0.05).
It is analyzed by ROC curve and calculates area under Receiver operating curve (AUC) to detect differential expression
The diagnosis performance of circRNA.The AUC of hsa_circ_0036847, hsa_circ_0064628, hsa_circ_0086569 distinguish
It is respectively 0.853-0.971,0.677-0.912,0.901-0.988 (Fig. 7-Fig. 9) for 0.906,0.853,0.961,95%CI.
The results show that 3 kinds of circRNA are used equally for the marker as diagnosis of colon cancer.
3 circRNA join together composition molecular marked compound express spectra AUC be 0.975, for colon cancer diagnosis imitate
Can be higher, further, it is also possible in conjunction with clinically common colon radiography, to further increase early screening and the prevention of colon cancer
Efficiency.
Above-described embodiment is the preferable embodiment of the disclosure, but embodiment of the present disclosure is not by above-described embodiment
It limits, made changes, modifications, substitutions, combinations, simplifications under other any spiritual essence and principles without departing from the disclosure,
It should be equivalent substitute mode, be included within the protection scope of the disclosure.
SEQUENCE LISTING
<110>Shandong Inst. of Tumor Prevention and Cure (Shandong Tumor Hospital)
<120>it is used for the marker of colon cancer screening
<130>
<160> 11
<170> PatentIn version 3.5
<210> 1
<211> 481
<212> DNA
<213> hsa_circ_0036847
<400> 1
gttcaatttc agaagcttca gcaactgcgc cttacacagt accatggagg atccttacca 60
aatgtgagcc agctgcggag cagtgcgtca gagtttcagc cgtcatttca ccaagctgat 120
aatgttcggg gaacccgcca tcacgggctg gtggagaggc catccaggaa ccgcttccac 180
cccctccacc gaaggtctgg ggacaagcca gggcgacaat ttgatggtag tgcttttgga 240
gccaattatt cctcacagcc tctggatgag agttggccaa ggcagcagcc tccttggaaa 300
gacgaaaagc atcctgggtt caggctgaca tctgcactta acaggaccaa ttctgattct 360
gctcttcaca cgagtgctct gagtaccaag ccccaggacc cctatggagg agggggccag 420
tcggcctggc ctgccccata catgggtttc tgtgatggtg agaataatgg acatggggaa 480
g 481
<210> 2
<211> 918
<212> DNA
<213> hsa_circ_0064628
<400> 2
ttgtcatgga tgcactggta gaagatgata tctgtattct gaatcatgaa aaagcccata 60
agagagatac agtgactcca gtttcaatat attcaggaga tgaatctgtt gcttcccatt 120
ttgctcttgt cactgcatat gaagacatca aaaaacgact taaggattca gagaaagaga 180
actctttgtt aaagaagaga ataagatttt tggaagaaaa gctaatagct cgatttgaag 240
aagaaacaag ttccgtggga cgagaacaag taaataaggc ctatcatgca tatcgagagg 300
tttgcattga tagagataat ttgaagagca aactggacaa aatgaataaa gacaactctg 360
aatctttgaa agtattgaat gagcagctac aatctaaaga agtagaactc ctccagctga 420
ggacagaggt ggaaactcag caggtgatga ggaatttaaa tccaccttca tcaaactggg 480
aggtggaaaa gttgagctgt gacctgaaga tccatggttt ggaacaagag ctggaactga 540
tgaggaaaga atgtagcgat ctcaaaatag aactacagaa agccaaacaa acggatccat 600
atcaggaaga caatctgaag agcagagatc tccaaaaact aagcatttca agacagagta 660
tctgtagcac agcctggagt gcagtggcgc aatcatggga tagcatgaat tttgaggact 720
gttccctctt cgcataagaa tagaagtagc catcagagtt catatacttt ttcaaccctt 780
tttcctcaaa tacgtatcat ttttacttaa gtaacttcaa ttctgtgttt taagctatct 840
aataattgtt gtttctacta tgttaagata gcctctctac attgttaccc tccatccaat 900
aaaaaatact gatctgac 918
<210> 3
<211> 226
<212> DNA
<213> hsa_circ_0086569
<400> 3
ggagagtccg gtagtgaaag gcaatgcgct gttagctcta agcagccttg ctgtcgtcgt 60
atctagacat gaagccagcc tctcctcaga ctctgacggg ctcctggagg ttcaacctaa 120
tttcctttca atgaaagagt gggtttccat ggtacttgat acactcttgg tcattgtgga 180
tagccattac caacccagag ggcaacttct ctcctggttt tattat 226
<210> 4
<211> 20
<212> DNA
<213>primer
<400> 4
agacgaaaag catcctgggt 20
<210> 5
<211> 21
<212> DNA
<213>primer
<400> 5
tctgaaattg aaccttcccc a 21
<210> 6
<211> 23
<212> DNA
<213>primer
<400> 6
tctacattgt taccctccat cca 23
<210> 7
<211> 21
<212> DNA
<213>primer
<400> 7
gcagtgacaa gagcaaaatg g 21
<210> 8
<211> 21
<212> DNA
<213>primer
<400> 8
ggatagccat taccaaccca g 21
<210> 9
<211> 18
<212> DNA
<213>primer
<400> 9
ggagcccgtc agagtctg 18
<210> 10
<211> 20
<212> DNA
<213>primer
<400> 10
ctcacagttg ccatgtagac 20
<210> 11
<211> 18
<212> DNA
<213>primer
<400> 11
gctcaattta tagaaacc 18
Claims (10)
- One or more of 1.hsa_circ_0036847, hsa_circ_0064628, hsa_circ_0086569 group cooperation For application of the marker in the product that preparation is used for colon cancer screening.
- 2. application according to claim 1, which is characterized in that the product for colon cancer screening includes detection patient's colon The reagent or medicine of hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_0086569 expression in tissue Object.
- 3. application according to claim 1, which is characterized in that the product for colon cancer screening includes: by fixed in real time It measures PCR, in situ hybridization, genetic chip or gene sequencing and detects hsa_circ_0036847, hsa_circ_0064628 or hsa_ Circ_0086569 expression is with the product of diagnosing colon.
- 4. application according to claim 1-3, which is characterized in that for the product of colon cancer screening, the product Hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_ in detection patient's colonic tissue can be passed through 0086569 expression carrys out screening colon cancer, testing result show hsa_circ_0036847, hsa_circ_0064628 or Hsa_circ_0086569 expression is significantly raised compared with normal tissue.
- 5. the fluorescence quantification PCR primer group of the circRNA marker for colon cancer screening, which is characterized in that the primer sets It is respectively used for amplifying hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_0086569.
- 6. fluorescence quantification PCR primer group according to claim 5, which is characterized in that for expanding hsa_circ_ 0036847 primer sets sequence includes SEQ ID NO:4 and SEQ ID NO:5；For expanding drawing for hsa_circ_0064628 Object group sequence includes SEQ ID NO:6 and SEQ ID NO:7；Primer sets sequence for expanding hsa_circ_0086569 includes SEQ ID NO:8 and SEQ ID NO:9.
- 7. the kit of the circRNA marker for colon cancer screening, including quantitative fluorescent PCR described in claim 5 or 6 Primer sets.
- 8. kit according to claim 7, which is characterized in that further include the primer sets for expanding reference gene；Preferably, reference gene is GAPDH；It is furthermore preferred that including SEQ ID NO:10 and SEQ ID NO:11 for expanding the primer sets sequence of GAPDH.
- 9. the method for colon cancer screening, which is characterized in that pass through hsa_circ_0036847, hsa_circ_0064628 or hsa_ The relative expression quantity of circ_0086569 judges to suffer from the risk of colon cancer；Preferably, pass through hsa_circ_0036847, hsa_circ_0064628 or hsa_ in detection patient's colonic tissue Circ_0086569 expression carrys out screening colon cancer, and testing result shows hsa_circ_0036847, hsa_circ_ 0064628 or hsa_circ_0086569 expression is significantly raised compared with normal tissue, then it represents that suffers from colon cancer；Preferably, the method also includes expanding to carry out screening in conjunction with colon radiography.
- One or more of 10.hsa_circ_0036847, hsa_circ_0064628 or hsa_circ_0086569 combination Application in the product of preparation colon cancer prognosis or treatment.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811641665.5A CN109457033B (en) | 2018-12-29 | 2018-12-29 | Marker for colon cancer screening |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811641665.5A CN109457033B (en) | 2018-12-29 | 2018-12-29 | Marker for colon cancer screening |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109457033A true CN109457033A (en) | 2019-03-12 |
| CN109457033B CN109457033B (en) | 2020-03-10 |
Family
ID=65615756
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811641665.5A Active CN109457033B (en) | 2018-12-29 | 2018-12-29 | Marker for colon cancer screening |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109457033B (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113493828A (en) * | 2021-07-12 | 2021-10-12 | 暨南大学 | Application of circular RNA in molecular marker of intestinal polyp |
Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2013144202A1 (en) * | 2012-03-29 | 2013-10-03 | Roche Diagnostics Gmbh | Biomarkers for discriminating healthy and/or non-malignant neoplastic colorectal cells from colorectal cancer cells |
| CN103667516A (en) * | 2014-01-07 | 2014-03-26 | 山东大学齐鲁医院 | miRNAs specific expression profile and diagnosis model for early colonic adenocarcinoma and rectal adenocarcinoma |
| CN103694332A (en) * | 2009-07-07 | 2014-04-02 | 清华大学 | New tumor marker |
| CN103966328A (en) * | 2014-05-07 | 2014-08-06 | 新乡医学院 | miRNA marker related to malignant transformation of colitis and proctitis and kit |
| CN105385752A (en) * | 2015-03-23 | 2016-03-09 | 复旦大学 | Detection method of human colon cancer protein marker Spondin-2 and detection kit thereof |
| CN105891484A (en) * | 2016-05-06 | 2016-08-24 | 中南大学湘雅医院 | Group of colon cancer metastasis related tumor stroma markers and application |
| CN106399464A (en) * | 2015-07-31 | 2017-02-15 | 复旦大学 | Human colorectal carcinoma molecular marker COL3A1 and application thereof |
| CN108624690A (en) * | 2018-06-19 | 2018-10-09 | 山东省疾病预防控制中心 | A kind of half adenocarcinoma of colon early screening detection kit of left and right and detection method |
| CN108753974A (en) * | 2018-06-27 | 2018-11-06 | 深圳华大生命科学研究院 | A kind of colorectal cancer tumor markers and its detection method and device |
| CN108866195A (en) * | 2018-08-17 | 2018-11-23 | 柳州市工人医院 | MiR-3942-5p is preparing the application in the general cancer diagnosis of damage-free type and prognosis drug as tumor markers |
-
2018
- 2018-12-29 CN CN201811641665.5A patent/CN109457033B/en active Active
Patent Citations (10)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103694332A (en) * | 2009-07-07 | 2014-04-02 | 清华大学 | New tumor marker |
| WO2013144202A1 (en) * | 2012-03-29 | 2013-10-03 | Roche Diagnostics Gmbh | Biomarkers for discriminating healthy and/or non-malignant neoplastic colorectal cells from colorectal cancer cells |
| CN103667516A (en) * | 2014-01-07 | 2014-03-26 | 山东大学齐鲁医院 | miRNAs specific expression profile and diagnosis model for early colonic adenocarcinoma and rectal adenocarcinoma |
| CN103966328A (en) * | 2014-05-07 | 2014-08-06 | 新乡医学院 | miRNA marker related to malignant transformation of colitis and proctitis and kit |
| CN105385752A (en) * | 2015-03-23 | 2016-03-09 | 复旦大学 | Detection method of human colon cancer protein marker Spondin-2 and detection kit thereof |
| CN106399464A (en) * | 2015-07-31 | 2017-02-15 | 复旦大学 | Human colorectal carcinoma molecular marker COL3A1 and application thereof |
| CN105891484A (en) * | 2016-05-06 | 2016-08-24 | 中南大学湘雅医院 | Group of colon cancer metastasis related tumor stroma markers and application |
| CN108624690A (en) * | 2018-06-19 | 2018-10-09 | 山东省疾病预防控制中心 | A kind of half adenocarcinoma of colon early screening detection kit of left and right and detection method |
| CN108753974A (en) * | 2018-06-27 | 2018-11-06 | 深圳华大生命科学研究院 | A kind of colorectal cancer tumor markers and its detection method and device |
| CN108866195A (en) * | 2018-08-17 | 2018-11-23 | 柳州市工人医院 | MiR-3942-5p is preparing the application in the general cancer diagnosis of damage-free type and prognosis drug as tumor markers |
Non-Patent Citations (1)
| Title |
|---|
| XIAOFENG WANG等: "The regulatory mechanism of Hsp90α secretion and its function in tumor malignancy", 《PNAS》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113493828A (en) * | 2021-07-12 | 2021-10-12 | 暨南大学 | Application of circular RNA in molecular marker of intestinal polyp |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109457033B (en) | 2020-03-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Huang et al. | Serum tRNA‐derived fragments (tRFs) as potential candidates for diagnosis of nontriple negative breast cancer | |
| CN108103198B (en) | One kind blood plasma miRNA marker relevant to cancer of pancreas auxiliary diagnosis and its application | |
| CN105950753B (en) | One kind blood plasma miRNA marker relevant to adenocarcinoma of lung auxiliary diagnosis and its application | |
| CN108624688B (en) | Application of hsa _ circ _0012755 as prostate cancer molecular target in preparation of medicines and kits | |
| CN106755344A (en) | Molecular marked compound and its application for the diagnosis of cancer of pancreas clinical prognosis | |
| CN109609633A (en) | One kind serum miRNA marker relevant to Computer-aided Diagnosis of Breast Cancer and its application | |
| CN109929932A (en) | Application in blood in two kinds of long-chain non-coding RNA Combining diagnosis esophageal squamous cell carcinomas | |
| CN109055557A (en) | One kind serum miRNA marker relevant to cancer of pancreas auxiliary diagnosis and its application | |
| CN108531586A (en) | A kind of relevant cycle miRNA marker and its application on X chromosome of and Computer-aided Diagnosis of Breast Cancer | |
| CN107881239A (en) | The miRNA marker related to colorectal cancer transfer and its application in blood plasma | |
| CN109609634A (en) | One kind circulation miRNA marker relevant to carcinoma of endometrium auxiliary diagnosis and its application | |
| CN106119347B (en) | The primer and kit of colorectal cancer transfer detection based on serum exosomal microRNAs | |
| CN109457033A (en) | Marker for colon cancer screening | |
| CN105219841B (en) | A kind of detection kit and its application of lung cancer differential expression microRNA | |
| CN109593851A (en) | One kind blood plasma miRNA marker relevant to Computer-aided Diagnosis of Breast Cancer and its application | |
| CN108929910A (en) | One kind serum miRNA marker relevant to adenocarcinoma of lung auxiliary diagnosis and its application | |
| CN108753981A (en) | Application of the quantitative detection of HOXB8 genes in colorectal cancer Index for diagnosis | |
| CN107858425A (en) | Applications and detection method of the miRNA 4741 as primary hepatic carcinoma diagnosis mark | |
| CN107746886B (en) | One kind blood plasma miRNA marker relevant to colorectal cancer auxiliary diagnosis and its application | |
| CN107723365B (en) | One kind blood plasma miRNA marker relevant to lung squamous cancer auxiliary diagnosis and its application | |
| CN110257514A (en) | A kind of new cancer of the esophagus blood miRNA marker and its application | |
| CN109022586A (en) | One kind blood plasma miRNA marker relevant to cervical carcinoma auxiliary diagnosis and its application | |
| CN111455057B (en) | Kit, device and method for lung cancer diagnosis | |
| CN107723366B (en) | One kind serum miRNA marker relevant to cardia cancer auxiliary diagnosis and its application | |
| CN116083583A (en) | Application of miRNA-483-5p in preparation of pancreatic cancer liver metastasis diagnosis kit |
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 |