CN102796820B - Hepatocellular carcinoma-related klotho gene single-nucleotide polymorphism and its construction method and use - Google Patents
Hepatocellular carcinoma-related klotho gene single-nucleotide polymorphism and its construction method and use Download PDFInfo
- Publication number
- CN102796820B CN102796820B CN2012102997487A CN201210299748A CN102796820B CN 102796820 B CN102796820 B CN 102796820B CN 2012102997487 A CN2012102997487 A CN 2012102997487A CN 201210299748 A CN201210299748 A CN 201210299748A CN 102796820 B CN102796820 B CN 102796820B
- Authority
- CN
- China
- Prior art keywords
- hepatocellular carcinoma
- klotho gene
- klotho
- polymorphism
- nucleotide polymorphism
- 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.)
- Expired - Fee Related
Links
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides hepatocellular carcinoma-related klotho gene single-nucleotide polymorphism and its construction method and use. The hepatocellular carcinoma-related klotho gene single-nucleotide polymorphism and its construction method have the advantages that 1, through a detection method provided by the invention, polymorphism of a single nucleotide located in an exon 4 area of a human klotho gene can be detected fast and simply and it is diagnosed that if an individual has susceptibility of hepatocellular carcinoma so that the hepatocellular carcinoma-susceptible population screening and hepatocellular carcinoma prevention are promoted; 2, a T allele point of rs648202 is used as a drug design target point for drug screening so that an active molecule adjusting a klotho gene expression level is selected and development of a novel antitumor drug is promoted; 3, through primer sequences and HaeIII endonuclease provided by the invention, simple, efficient and specific detection of rs648202 polymorphism is realized; and 4, according to hepatocellular carcinoma-related klotho gene single-nucleotide polymorphism, a hepatocellular carcinoma genetic diagnosis kit can be constructed.
Description
Technical field
The invention provides its construction process of klotho gene mononucleotide polymorphism and application thereof that a kind of primary hepatocellular carcinoma (hcc) is relevant, belong to the genetically engineered biological technical field.
Background technology
Primary hepatocellular carcinoma (hcc) (is called for short liver cancer, HCC) be one of modal malignant tumour in world wide, the data presentation of in December, 2007 ACS (ACS) issue: increase 66.7 ten thousand examples liver cancer year, dead 59.8 ten thousand examples, wherein approximately 50% new trouble case and death in China, be that liver cancer is larger to China people's health threat, become China second tumour killer, not only make the patient suffering can't bear, survival rate is low, and brings serious economy and psychological burden to family, society.
Single nucleotide polymorphism (SNP) refers on genomic level the polymorphism that the variation by the single core thuja acid causes.It is modal a kind of in the heritable variation of the mankind, accounts for more than 90% of all known polymorphisms.SNP is as the early stage sudden change of genetic stability, closely related with disease.Apparently higher than non-patient, point out this SNP disease-related therewith when the frequency of occurrences of a kind of SNP in the patient, by analyzing and researching both haplotype and linkage disequilibrium, the Disease-causing gene of any the unknown in genome can be located.The effect of SNP performance in the Disease-causing gene location mainly comprises: the one, finding the SNP that causes a disease, the appearance of this SNP can cause gene expression amount and (or) variation of protein expressioning product structure, thereby cause the generation of certain disease or make individual to certain disease-susceptible humans; The 2nd, SNP is as a kind of genetic marker, chain with disease or phenotype.
Klotho finds the gene relevant with aging in 1997, is positioned at human chromosomal 13q12, and length is about 50kb, 5 exons and 4 introns, consists of.The expression deletion of Klotho gene in mouse causes its various phenotypes that the mankind aging occurs being similar to, as arteriosclerosis, osteoporosis, pulmonary emphysema, the lost of life, skin atrophy, infertility, ataxia etc.In recent years, increasing research discovery klotho gene can be regulated IGF, Wnt, the signal paths such as TGF-β l, FGF, participate in the oxidative stress process, and relevant with the immune deficiency of body, the vital role of the similar cancer suppressor gene of performance in the carcinogenesis of human of malignant tumour.Therefore can utilize that klotho is used as that tumour occurs, the mark of classification and as target spot for antineoplastic treatment.
Domestic and foreign literature and patent to prior art are retrieved, the relevant report of so far there are no any klotho gene pleiomorphism and hepatocellular carcinoma susceptibility relation.
Summary of the invention
The object of the invention is to fill up the blank of prior art, its construction process of klotho gene mononucleotide polymorphism and application thereof that a kind of primary hepatocellular carcinoma (hcc) is relevant are provided.
The invention provides a kind of method that detects the klotho gene mononucleotide polymorphism that primary hepatocellular carcinoma (hcc) is relevant, adopt the PCR-RFLP method, comprise the following steps:
(1) take the people's complete genome DNA to be measured that comprises the klotho gene is template, and pcr amplification obtains the DNA sequence dna that comprises the rs648202 mononucleotide polymorphic in klotho gene extron 4th district, i.e. sequence shown in the 2190th to the 2352nd in SEQ ID No.1,
Wherein, the PCR primer sequence is as follows:
Upstream primer is F:5 '-ATAGCCTTGCAGGCTGATTG-3 '
Downstream primer is R:5 '-TTCTTTGGTTCAGCCAGTCC-3 '
(2) pcr amplification product is carried out after enzyme is cut carrying out the rs648202 single nucleotide polymorphism typing by agarose gel electrophoresis through restriction enzyme Hae III, the DNA fragmentation length after CC genotype enzyme is cut is still 163bp; CT genotype enzyme is cut the rear DNA fragmentation that produces 163bp, 100bp, tri-kinds of length of 63bp; TT genotype enzyme is cut the DNA fragmentation of rear generation 100bp, bis-kinds of length of 63bp.
The present invention also provides a kind of isolating nucleic acid, have sequence shown in the 2190th to the 2352nd in SEQ ID No.1, and the base of the 2255th in SEQ ID No.1 is T.
The present invention also provides a kind of Auele Specific Primer that detects the klotho gene mononucleotide polymorphism that primary hepatocellular carcinoma (hcc) is relevant, upstream primer is F:5 '-ATAGCCTTGCAGGCTGATTG-3 ', and downstream primer is R:5 '-TTCTTTGGTTCAGCCAGTCC-3 '.
The present invention also provides a kind of diagnostic kit of primary hepatocyte hepatocarcinoma susceptibility, and it comprises:
(1) primer of specific amplification klotho gene mononucleotide polymorphism rs648202, wherein, upstream primer is F:5 '-ATAGCCTTGCAGGCTGATTG-3 ', downstream primer is R:5 '-TTCTTTGGTTCAGCCAGTCC-3 ';
(2) detect amplified production and compare the reagent that whether exists variation required with normal klotho gene.
Utilize the nucleotide sequence of the pleomorphism site relevant to primary hepatocyte hepatocarcinoma provided by the present invention, can build the test kit that primary hepatocyte hepatocarcinoma Susceptible population is carried out to genetic screening, target spot using it as medicinal design, find out the bioactive molecule with adjusting klotho genetic expression, thereby promote the discovery of new antitumor drug.
The present invention has following technique effect:
(1) method that the present invention sets up can detect the genotype that is positioned at human chromosomal 13q12 zone klotho gene extron 4 district mononucleotide polymorphism site rs648202 quickly and easily, judge by detecting T allelic having or not whether individuality has susceptibility to primary hepatocyte hepatocarcinoma, thereby be conducive to the examination of primary hepatocyte hepatocarcinoma Susceptible population and the prevention of primary hepatocyte hepatocarcinoma;
(2) utilize the T loci of rs648202 to carry out the screening of medicine as the medicinal design target spot, filter out the bioactive molecule that can regulate the klotho gene expression dose, promote the discovery of new antitumor drug;
(3) utilize primer sequence provided by the invention and Hae III endonuclease to detect easy, efficiently, specifically rs648202 polymorphic;
(4) utilize the klotho gene mononucleotide polymorphism that primary hepatocellular carcinoma (hcc) provided by the invention is relevant, build the test kit that primary hepatocyte hepatocarcinoma is carried out to the heredity diagnosis;
(5) because klotho has participated in malignant proliferation, apoptosis, pathologic process that invasion and attack are relevant to cellular activities such as transfers, so the present invention provides experience and application foundation to the relation of further investigated klotho and other diseases from now on.
The accompanying drawing explanation
Figure l is for being used Auele Specific Primer to carry out the agarose gel electrophoresis figure after pcr amplification, Hae III enzyme are cut to klotho gene extron 4th district.M:DNA molecular weight standard wherein; The l:CT genotype; The 2:CC genotype; The 3:TT genotype; 4: blank.
Embodiment
Further illustrate the present invention below in conjunction with specific embodiment.Should be noted, following examples, only for the present invention is described, limit the scope of the invention and be not used in.
(1) collect blood sample and DNA extraction
Collect sporadic primary hepatocyte hepatocarcinoma clinical samples 212 examples from The Second Affiliated Hospital of Nanjing Medical University, all patients all make a definite diagnosis through histopathology, male 150 examples wherein, female's 62 examples.Patient's mean age is 62 years old; Control group 288 examples, the mean age is 61 years old.The age of case group and control group, sex form there was no significant difference (P>0.05).Above result show two groups aspect age, sex equilibrium comparable.All research objects
All signaturesInformed Consent Form.
Use ordinary method to isolate white corpuscle from the above-mentioned blood sample of collecting, and adopt classical
PhenolAfter-chloroform method extracting white corpuscle genomic dna, the sample unification is diluted to 0.1 μ g/ μ l, as DNA profiling, remaining DNA is positioned over-20 ℃ of preservations.
(2) design and synthesize nucleotide primer, primer sequence is as follows,
Upstream primer is F:5 '-ATAGCCTTGCAGGCTGATTG-3 ',
Downstream primer is R:5 '-TTCTTTGGTTCAGCCAGTCC-3 '.
Carry out polymerase chain reaction (PCR)
Carry out the PCR reaction system: total amount is 20 μ l
Reaction conditions is set in the pcr amplification instrument:
What obtain klotho gene extron 4 district 163bp after final amplification comprises the polymorphic DNA sequence dna of rs648202 mononucleotide, i.e. sequence shown in the 2190th to the 2352nd in SEQ ID No.1.
(3) DNA sequence dna amplification obtained carries out the single nucleotide polymorphism somatotype
To the DNA fragmentation product obtained in step (2), utilize restriction endonuclease to carry out the restriction fragment length polymorphism somatotype.
Concrete grammar is: the restriction enzyme HaeIII and the corresponding 10 * enzyme that add 5U in 5 μ l pcr amplification products
Cut reactionDamping fluid 1.0 μ l, add water to 10 μ l, after mixing, under 37 ℃, digests 6 hours.
Getting enzyme cutsProduct 10 μ l add 1.0 μ l
The loading bufferingLiquid, fully mix, the well that adds 3% sepharose, do reference with DNA Marker, the electrophoresis working fluid is 1 * TBE, and sample is moved to positive pole by negative pole, constant voltage 80V approximately will coagulate the chamber plate and be placed on ultraviolet transilluminator silica glass platform and detected after 40 minutes, observe each swimming lane and fluorescence band whether occurs, gel images analyser enzyme analysis is cut the genotype type of product, takes pictures and records enzyme and cut result.As shown in Figure 1, the DNA fragmentation length of CC genotype enzyme after cutting is still 163bp; CT genotype enzyme is cut the rear DNA fragmentation that produces 163bp, 100bp, tri-kinds of length of 63bp; TT genotype enzyme is cut the DNA fragmentation of rear generation 100bp, bis-kinds of length of 63bp.
(4) correlation analysis between Klotho coding region rs648202 mononucleotide polymorphic and primary hepatocyte hepatocarcinoma genetic predisposition
Statistical method: adopt SPSS11.0 software building database.Allelotrope and genotypic frequency to the klotho gene of case and contrast
The distribution state property andDemographic variables, smoking and the variable such as drink adopt χ
2Whether check, exist genotype or allelic distribution bias to judge selected crowd, thereby determine its representativeness to whole crowd.Obtain odds ratio (OR) and 95% credibility interval (CI), all OR with single argument logistic regression analysis
Value is through sexWith the age, correct.Statistical study SAS9.0 software, P<0.05 is for statistics has significant difference, and all checks are two-tailed test.
Statistics is as follows:
1. the genetic equilibrium of Klotho gene rs648202 loci gene type frequency check
Klotho gene rs648202 loci gene type frequency is carried out to the check of Hardy-Weniberg genetic equilibrium, χ
2=2.272, P=0.132, difference not statistically significant between this locus gene frequency observed value and expected value, show that this genotype frequency all meets the Hardy-Weniberg ratio, sample has colony's representativeness.
2. the rs648202 polymorphic site genotype between primary hepatocyte hepatocarcinoma patient group and normal healthy controls group and loci distribution frequency are as shown in table 1 below:
The genotype that table 1Klotho gene rs648202 is polymorphic and gene frequency and with the relation of liver cancer
CC, CT and TT genotype frequency are respectively 57.1%37.7% and 5.2% in case, in contrast, are respectively 68.7%, 27.1% and 4.2%, and genotypic distributional difference has significant statistical significance (P=0.026).GA and AA merging are compared with GG, and both still have significant statistical significance (P=0.007).The Logistic regression analysis shows, with carrying the CC wild-type, compares, and the genotypic individual danger that liver cancer occurs of CT of carrying variation significantly raises, and increased value is 59.6%, and the danger of carrying CT+TT mutator gene type person increases by 60.4%.
These results suggest, klotho gene extron 4 district rs648202 are significant sites to the primary hepatocyte hepatocarcinoma susceptible.When this site is carried T allelotrope the genotype in this site is T/T or C/T, the individuality that its individual danger that liver cancer occurs is C/C apparently higher than genotype, show that the genotypic individuality of C/T or T/T is to the primary hepatocyte hepatocarcinoma susceptible, and the genotypic individuality of C/C is to primary hepatocyte hepatocarcinoma susceptible not comparatively.
Organization Applicant
----------------------
Street :
City :
State :
Country :
PostalCode :
PhoneNumber :
FaxNumber :
EmailAddress :
<110 > OrganizationName: The Second Affiliated Hospital of Nanjing Medical University
Individual Applicant
--------------------
Street :
City :
State :
Country :
PostalCode :
PhoneNumber :
FaxNumber :
EmailAddress :
<110 > LastName: Huang
<110 > FirstName: daybreak
<110> MiddleInitial :
<110> Suffix :
Application Project
-------------------
<120 > Title: its construction process of klotho gene mononucleotide polymorphism that a kind of primary hepatocellular carcinoma (hcc) is relevant and
Application
<130> AppFileReference :
<140> CurrentAppNumber :
<141> CurrentFilingDate : ____-__-__
Sequence
--------
<213> OrganismName :
<400> PreSequenceString :
cgcgcagcat gcccgccagc gccccgccgc gccgcccgcg gccgccgccg ccgtcgctgt 60
cgctgctgct ggtgctgctg ggcctgggcg gccgccgcct gcgtgcggag ccgggcgacg 120
gcgcgcagac ctgggcccgt ttctcgcggc ctcctgcccc cgaggccgcg ggcctcttcc 180
agggcacctt ccccgacggc ttcctctggg ccgtgggcag cgccgcctac cagaccgagg 240
gcggctggca gcagcacggc aagggtgcgt ccatctggga tacgttcacc caccaccccc 300
tggcaccccc gggagactcc cggaacgcca gtctgccgtt gggcgccccg tcgccgctgc 360
agcccgccac cggggacgta gccagcgaca gctacaacaa cgtcttccgc gacacggagg 420
cgctgcgcga gctcggggtc actcactacc gcttctccat ctcgtgggcg cgagtgctcc 480
ccaatggcag cgcgggcgtc cccaaccgcg aggggctgcg ctactaccgg cgcctgctgg 540
agcggctgcg ggagctgggc gtgcagcccg tggtcaccct gtaccactgg gacctgcccc 600
agcgcctgca ggacgcctac ggcggctggg ccaaccgcgc cctggccgac cacttcaggg 660
attacgcgga gctctgcttc cgccacttcg gcggtcaggt caagtactgg atcaccatcg 720
acaaccccta cgtggtggcc tggcacggct acgccaccgg gcgcctggcc cccggcatcc 780
ggggcagccc gcggctcggg tacctggtgg cgcacaacct cctcctggct catgccaaag 840
tctggcatct ctacaatact tctttccgtc ccactcaggg aggtcaggtg tccattgccc 900
taagctctca ctggatcaat cctcgaagaa tgaccgacca cagcatcaaa gaatgtcaaa 960
aatctctgga ctttgtacta ggttggtttg ccaaacccgt atttattgat ggtgactatc 1020
ccgagagcat gaagaataac ctttcatcta ttctgcctga ttttactgaa tctgagaaaa 1080
agttcatcaa aggaactgct gacttttttg ctctttgctt tggacccacc ttgagttttc 1140
aacttttgga ccctcacatg aagttccgcc aattggaatc tcccaacctg aggcaactgc 1200
tttcctggat tgaccttgaa tttaaccatc ctcaaatatt tattgtggaa aatggctggt 1260
ttgtctcagg gaccaccaag agagatgatg ccaaatatat gtattacctc aaaaagttca 1320
tcatggaaac cttaaaagcc atcaagctgg atggggtgga tgtcatcggg tataccgcat 1380
ggtccctcat ggatggtttc gagtggcaca gaggttacag catcaggcgt ggactcttct 1440
atgttgactt tctaagccag gacaagatgt tgttgccaaa gtcttcagcc ttgttctacc 1500
aaaagctgat agagaaaaat ggcttccctc ctttacctga aaatcagccc ctagaaggga 1560
catttccctg tgactttgct tggggagttg ttgacaacta cattcaagta gataccactc 1620
tgtctcagtt taccgacctg aatgtttacc tgtgggatgt ccaccacagt aaaaggctta 1680
ttaaagtgga tggggttgtg accaagaaga ggaaatccta ctgtgttgac tttgctgcca 1740
tccagcccca gatcgcttta ctccaggaaa tgcacgttac acattttcgc ttctccctgg 1800
actgggccct gattctccct ctgggtaacc agtcccaggt gaaccacacc atcctgcagt 1860
actatcgctg catggccagc gagcttgtcc gtgtcaacat caccccagtg gtggccctgt 1920
ggcagcctat ggccccgaac caaggactgc cgcgcctcct ggccaggcag ggcgcctggg 1980
agaaccccta cactgccctg gcctttgcag agtatgcccg actgtgcttt caagagctcg 2040
gccatcacgt caagctttgg ataacgatga atgagccgta tacaaggaat atgacataca 2100
gtgctggcca caaccttctg aaggcccatg ccctggcttg gcatgtgtac aatgaaaagt 2160
ttaggcatgc tcagaatggg aaaatatcca tagccttgca ggctgattgg atagaacctg 2220
cctgcccttt ctcccaaaag gacaaagagg tggctgagag agttttggaa tttgacattg 2280
gctggctggc tgagcccatt ttcggctctg gagattatcc atgggtgatg agggactggc 2340
tgaaccaaag aaacaatttt cttcttcctt atttcactga agatgaaaaa aagctaatcc 2400
agggtacctt tgactttttg gctttaagcc attataccac catccttgta gactcagaaa 2460
aagaagatcc aataaaatac aatgattacc tagaagtgca agaaatgacc gacatcacgt 2520
ggctcaactc ccccagtcag gtggcggtag tgccctgggg gttgcgcaaa gtgctgaact 2580
ggctgaagtt caagtacgga gacctcccca tgtacataat atccaatgga atcgatgacg 2640
ggctgcatgc tgaggacgac cagctgaggg tgtattatat gcagaattac ataaacgaag 2700
ctctcaaagc ccacatactg gatggtatca atctttgcgg atactttgct tattcgttta 2760
acgaccgcac agctccgagg tttggcctct atcgttatgc tgcagatcag tttgagccca 2820
aggcatccat gaaacattac aggaaaatta ttgacagcaa tggtttcccg ggcccagaaa 2880
ctctggaaag attttgtcca gaagaattca ccgtgtgtac tgagtgcagt ttttttcaca 2940
cccgaaagtc tttactggct ttcatagctt ttctattttt tgcttctatt atttctctct 3000
cccttatatt ttactactcg aagaaaggca gaagaagtta caaatagttc tgaacatttt 3060
tctattcatt cattttgaaa taattatgca gacacatcag ctgttaacca tttgcacctc 3120
taagtgttgt gaaactgtaa atttcataca tttgacttct agaaaacatt tttgtggctt 3180
atgacagagg ttttgaaatg ggcataggtg atcgtaaaat attgaataat gcgaatagtg 3240
cctgaatttg ttctcttttt gggtgattaa aaaactgaca ggcactataa tttctgtaac 3300
acactaacaa aagcatgaaa aataggaacc acaccaatgc aacatttgtg cagaaatttg 3360
aatgacaaga ttaggaatat tttcttctgc acccacttct aaatttaatg tttttctgga 3420
agtagtaatt gcaagagttc gaatagaaag ttatgtacca agtaaccatt tctcagctgc 3480
cataataatg cctagtggct tcccctctgt caaatctagt ttcctatgga aaagaagatg 3540
gcagatacag gagagacgac agagggtcct aggctggaat gttcctttcg aaagcaatgc 3600
ttctatcaaa tactagtatt aatttatgta tctggttaat gacatacttg gagagcaaat 3660
tatggaaatg tgtattttat atgatttttg aggtcctgtc taaaccctgt gtccctgagg 3720
gatctgtctc actggcatct tgttgagggc cttgcacata ggaaactttt gataagtatc 3780
tgcggaaaaa caaacatgaa tcctgtgata ttgggctctt caggaagcat aaagcaattg 3840
tgaaatacag tataccgcag tggctctagg tggaggaaag gaggaaaaag tgcttattat 3900
gtgcaacatt atgattaatc tgattataca ccatttttga gcagatcttg gaatgaatga 3960
catgaccttt ccctagagaa taaggatgaa ataatcactc attctatgaa cagtgacact 4020
actttctatt ctttagctgt actgtaattt ctttgagttg atagttttac aaattcttaa 4080
taggttcaaa agcaatctgg tctgaataac actggatttg tttctgtgat ctctgaggtc 4140
tattttatgt ttttgctgct acttctgtgg aagtagcttt gaactagttt tactttgaac 4200
tttcacgctg aaacatgcta gtgatatcta gaaagggcta attaggtctc atcctttaat 4260
gccccttaaa taagtcttgc tgattttcag acagggaagt ctctctatta cactggagct 4320
gttttataga taagtcaata ttgtatcagg caagataaac caatgtcata acaggcattg 4380
ccaacctcac tgacacaggg tcatagtgta taataatata ctgtactata taatatatca 4440
tctttagagg tatgattttt tcatgaaaga taagcttttg gtaatattca ttttaaagtg 4500
gacttattaa aattggatgc tagagaatca agtttatttt atgtatatat ttttctgatt 4560
ataagagtaa tatatgttca ttgtaaaaat ttttaaaaca cagaaactat atgcaaagaa 4620
aaaataaaaa ttatctataa tctcagaacc cagaaatagc cactattaac atttcctacg 4680
tattttattt tacatagatc atattgtata tagttagtat ctttattaat ttttattatg 4740
aaactttcct ttgtcattat tagtcttcaa aagcatgatt tttaatagtt gttgagtatt 4800
ccaccacagg aatgtatcac aacttaaccg ttcccgtttg ttagactagt ttcttattaa 4860
tgttgatgaa tgttgtttaa aaataatttt gttgctacat ttactttaat ttccttgact 4920
gtaaagagaa gtaattttgc tccttgataa agtattatat taataataaa tctgcctgca 4980
actttttgcc ttctttcata atcataaaaa aa 5012
<212> Type : DNA
<211> Length : 5012
SequenceName : 1
SequenceDescription :
Claims (1)
1. the purposes of a species-specific primer on the diagnostic kit for preparing the primary hepatocellular carcinoma (hcc) susceptibility, it is characterized in that, described Auele Specific Primer, upstream primer is F:5'-ATAGCCTTGCAGGCTGATTG-3', downstream primer is R:5'-TTCTTTGGTTCAGCCAGTCC-3', described diagnostic kit, for detection of the genotype in klotho gene rs648202 site, detects and adopts the PCR-RFLP method, comprises the following steps:
(1) the people's complete genome DNA to be measured that comprises the klotho gene of take is template, carry out with above-mentioned Auele Specific Primer the DNA sequence dna that comprises the rs648202 mononucleotide polymorphic that pcr amplification obtains klotho gene extron 4th district, i.e. sequence shown in the 2190th to the 2352nd in SEQ ID No.1;
(2) pcr amplification product is carried out after enzyme is cut carrying out the rs648202 single nucleotide polymorphism typing by agarose gel electrophoresis through restriction enzyme Hae III.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2012102997487A CN102796820B (en) | 2012-08-22 | 2012-08-22 | Hepatocellular carcinoma-related klotho gene single-nucleotide polymorphism and its construction method and use |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN2012102997487A CN102796820B (en) | 2012-08-22 | 2012-08-22 | Hepatocellular carcinoma-related klotho gene single-nucleotide polymorphism and its construction method and use |
Publications (2)
Publication Number | Publication Date |
---|---|
CN102796820A CN102796820A (en) | 2012-11-28 |
CN102796820B true CN102796820B (en) | 2013-12-04 |
Family
ID=47196117
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN2012102997487A Expired - Fee Related CN102796820B (en) | 2012-08-22 | 2012-08-22 | Hepatocellular carcinoma-related klotho gene single-nucleotide polymorphism and its construction method and use |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN102796820B (en) |
Families Citing this family (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN102961739A (en) * | 2012-12-12 | 2013-03-13 | 黄曙 | Application of KLOTHO protein |
CN107312829A (en) * | 2017-04-24 | 2017-11-03 | 首都医科大学附属北京佑安医院 | The mononucleotide polymorphism site related from different recurrence possibility liver cancer and its application |
CN107312830B (en) * | 2017-04-24 | 2021-04-13 | 首都医科大学附属北京佑安医院 | Single nucleotide polymorphism sites related to liver cancers with different degrees of malignancy and application thereof |
CN106957921B (en) * | 2017-05-18 | 2020-12-29 | 中国水产科学研究院珠江水产研究所 | SNP locus suitable for klotho gene of micropterus salmoides fed with artificial compound feed and application of SNP locus in breeding |
CN107557461B (en) * | 2017-10-20 | 2021-02-19 | 武汉赛云博生物科技有限公司 | Detection method of nucleic acid mass spectrum for early screening of liver cancer susceptibility genes |
CN108333355B (en) * | 2018-02-01 | 2020-05-26 | 黄曙 | Application of KLOTHO protein in preparation of reagent for diagnosing gastrointestinal stromal tumor risk |
CN112915192B (en) * | 2021-03-05 | 2022-10-25 | 南方医科大学南方医院 | Application of KP-1 in preparation of medicine for treating chronic liver diseases |
CN113444730A (en) * | 2021-03-17 | 2021-09-28 | 昆明市延安医院 | Screening and constructing method of primary hepatocyte klotho gene transduction stem cells |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2152295B1 (en) * | 2007-05-08 | 2018-04-18 | Tel HaShomer Medical Research Infrastructure and Services Ltd. | Klotho protein and related compounds for the treatment of cancer |
CA2718869C (en) * | 2008-03-19 | 2018-10-30 | China Synthetic Rubber Corporation | Methods and agents for the diagnosis and treatment of hepatocellular carcinoma |
CN101608217A (en) * | 2008-06-20 | 2009-12-23 | 上海主健生物工程有限公司 | Liver cancer genetic test kit |
US20120014927A1 (en) * | 2008-12-03 | 2012-01-19 | Tel Hashomer Medical Research Infrastructure And Services Ltd. | Methods and kits for determining predisposition to cancer |
EP2388336A1 (en) * | 2010-05-19 | 2011-11-23 | Signature Diagnostics AG | Method and kits for diagnosing colorectal cancer |
-
2012
- 2012-08-22 CN CN2012102997487A patent/CN102796820B/en not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
---|---|
CN102796820A (en) | 2012-11-28 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN102796820B (en) | Hepatocellular carcinoma-related klotho gene single-nucleotide polymorphism and its construction method and use | |
CN103436606B (en) | Kit for auxiliary diagnosis and/or prognosis judgment of esophageal carcinoma | |
Shih et al. | Association of TNF-α polymorphism with susceptibility to and severity of non-small cell lung cancer | |
Velickovic et al. | Intragenic PTEN/MMAC1 loss of heterozygosity in conventional (clear-cell) renal cell carcinoma is associated with poor patient prognosis | |
Karlsson et al. | MAGI1 copy number variation in bipolar affective disorder and schizophrenia | |
CN103911452B (en) | Chinese population deaf gene screening kit and application thereof | |
CN101298629B (en) | Application of LRRC4 gene promoter zone methylation detection to glioma diagnosis and detection system | |
de Andrade et al. | Chromoblastomycosis in the Amazon region, Brazil, caused by Fonsecaea pedrosoi, Fonsecaea nubica, and Rhinocladiella similis: Clinicopathology, susceptibility, and molecular identification | |
Yuan et al. | Genetic profile characterization and population study of 21 autosomal STR in C hinese K azak ethnic minority group | |
Lu et al. | Additional genomic duplications in AZFc underlie the b2/b3 deletion-associated risk of spermatogenic impairment in Han Chinese population | |
CN109182527B (en) | Interferon related kit for prognosis evaluation and chemotherapy effect prediction in glioma | |
CN110283908A (en) | A kind of colorectal cancer auxiliary diagnosis SNP marker and its application | |
Xue et al. | Indirubin attenuates mouse psoriasis-like skin lesion in a CD274-dependent manner: an achievement of RNA sequencing | |
Krueger et al. | Secreted toxins from Staphylococcus aureus strains isolated from keratinocyte skin cancers mediate pro-tumorigenic inflammatory responses in the skin | |
CN107254546B (en) | SNP marker related to breast cancer neoadjuvant chemotherapy curative effect and application thereof | |
He et al. | Mutation in the hair cell specific gene POU4F3 is a common cause for autosomal dominant nonsyndromic hearing loss in Chinese Hans | |
Mo et al. | Positive association between IL-16 rs11556218 T/G polymorphism and cancer risk: a meta-analysis | |
CN101250589B (en) | Reagent case for detecting POU3F4 gene 499C>T mutation | |
CN102304584A (en) | GJB2 gene p.V37I mutation diagnostic kit for diagnosing late-onset deafness | |
Xavier et al. | Gene expression profiling and association studies implicate the neuregulin signaling pathway in Behcet's disease susceptibility | |
CN103642921A (en) | Selection, detection and application of catalase gene tagging single nucleotide polymorphic sites | |
CN104099338B (en) | MYO15A gene mutation body and its application | |
CN105838720A (en) | PTPRQ gene mutant and application thereof | |
CN103602671A (en) | SNP in KRT8 gene exon area and determination method thereof | |
Rasi et al. | Saliva is a reliable and practical source of germline DNA for genome-wide studies in chronic lymphocytic leukemia |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
C06 | Publication | ||
PB01 | Publication | ||
C10 | Entry into substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
C14 | Grant of patent or utility model | ||
GR01 | Patent grant | ||
CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20131204 Termination date: 20150822 |
|
EXPY | Termination of patent right or utility model |