CN109679957A - IncRNALNC_004208 and its detection reagent are preparing the application in glioma prognostic agent - Google Patents
IncRNALNC_004208 and its detection reagent are preparing the application in glioma prognostic agent Download PDFInfo
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Abstract
The invention discloses a kind of IncRNALNC_004208 and its detection reagent to prepare the application in glioma prognostic agent.The present invention passes through research and utilization lncRNA sequencing discovery, lncRNA LNC_004208 is in overexpression in the brain glioblastoma cell system that Temozolomide is resisted, by having carried out Effect of follow-up visit by telephone to 104 Patients with gliomas, their start time has been inquired in detail, rank, treatment condition, whether there is or not recurrences, whether there is or not suffer from other diseases again, whether there is or not use other drugs, recurrence and death time etc., and register life span and state, it was found that the patient that high expression patient's mean survival time and progression free survival phase are significantly shorter than low expression or do not express, and it is related with the reactivity of Temozolomide.Illustrate that LNC_004208 is molecular labeling relevant to glioma prognosis, lncRNA LNC_004208 expression is high, patient's poor prognosis.
Description
Technical field
The invention belongs to oncomolecularbiology fields, and in particular to a kind of IncRNALNC_004208 and its detection reagent
Preparing application and corresponding reagent in glioma prognostic agent.
Background technique
Glioma is the most common malignant tumour of central nervous system, accounts for about the 80% of primary malignancy brain tumor, root
It is counted according to European neural tumor association (EANO), glioma every Annual occurence rate in the whole world is about 6/100,000.Glioma
Treatment method is global problem, especially malignant glioblastoma, even if taking optimum treatment scheme, patient's prognosis is still
It is old very undesirable, mean survival time (MST) only 13.3-15 months.Temozolomide (Temozolomide, TMZ) is to go out in the latest 20 years
It is existing it is a kind of treat the preferable new drug of glioma effect, recommend through U.S. FDA approval, the comprehensive cancer network how-tos of US National
Have become the preferred chemotherapeutics for the treatment of glioma.TMZ belongs to novel imidazole piperazine class and takes orally antineoplastic, is easy to through blood tumor
Barrier, activated product MTIC are played cytotoxicity and are made by N3, N7, O6 guanines and the alkylation of O3 adenine dnas
With.Due in tumour cell factor or acquired resistance appearance, Patients with gliomas to TMZ treatment gradually produce
It resists, oncotherapy curative effect reduces.
O6-Methylguanine DNA-Methyltransferase (MGMT) is by O6 methyl abnormal on transfer DNA, to inhibit
The DNA damage that TMZ is mediated causes glioma cell TMZ to resist.It recent studies have shown that, other than MGMT, tumour cell epidermis
The exception of many A signal pathways such as growth factor (epidermal growth factor receptor, EGFR) both participates in
TMZ resist generation, however its specific mechanism has not yet been elucidated.Therefore, further investigated TMZ resists the molecular mechanism occurred, and adopts
It takes effective measures to be intervened, important new approaches will be provided for the chemotherapeutic efficacy for improving glioma.
Recent study finds that lncRNAs express spectra has significant difference, brain glue in different classes of glioma cells in tissue
The morbidity and prognosis of matter tumor and the expression of lncRNAs and functional disturbance are closely related.Molecular pathology is in accurate Medical Era
Extremely important effect is increasingly played in tumor research and diagnosis and treatment.With tumor development Mechanism Study and targeted drug
The progress at full speed of exploitation and clinical application, tumor-related molecules pathological examination also from tissue or cytologic specimen, extend to liquid
Body biopsy；The analysis means of use are also more sensitive, stage construction and high throughput, and gradually stride forward to intelligence.Accurate molecule point
Type has become indispensable link during diagnosing tumor, treatment and process management.Especially across tumour happening part, with
Exploration and practice of the genotype as the treatment mode of direct target, puts forward new requirements to molecular pathology.
Summary of the invention
The first purpose of the invention is to provide a kind of IncRNALNC_004208, sequence is shown in SEQ NO:1.The present invention with
LncRNA LNC_004208 is Diagnosing Gliomas, prognosis and the treatment new strategy of target, is research lncRNAs in brain glue
Molecular mechanism in matter tumor TMZ resistance provides new experimental evidence.
A second object of the present invention is to provide a kind of reagents for detecting above-mentioned IncRNALNC_004208 expression quantity to make
Application in standby glioma prognosis or detection TMZ sensitive formulation.
Further, the reagent of the detection IncRNA LNC_004208 expression quantity is for detecting glioma cells in tissue.
Further, the reagent of the detection IncRNALNC_004208 expression quantity includes real time fluorescent quantitative detection examination
Agent.
Primer for real time fluorescent quantitative detection lncRNA LNC_004208 expression quantity includes following two pairs: (according to not
It with the primer of Position Design, can all use, one pair of them can also be selected)
LNC_004208 (1)-forward primer 5 '-GCCAATCCAAACCGAAAGCC-3 ', is shown in SEQ NO:2；
LNC_004208 (1)-reverse primer 5 '-AGACTCAGACCCCAGGCTAC-3 ', is shown in SEQ NO:3；
LNC_004208 (2)-forward primer 5 '-TTTGCTACATCCCAGCTCCA-3 ', is shown in SEQ NO:4；
LNC_004208 (2)-reverse primer 5 '-TCCCCCTTTCCCTTGTAGGT-3 ', is shown in SEQ NO:5.
Third object of the present invention is to provide a kind of glioma prognosis or detection TMZ sensibility reagents, should
The sequence of IncRNALNC_004208 is shown in SEQ NO:1.
The glioma prognosis or detection TMZ sensibility reagent, for detection IncRNALNC_004208 expression quantity
Reagent.
The reagent of the detection IncRNA LNC_004208 expression quantity includes real time fluorescent quantitative detection reagent.
Primer for real time fluorescent quantitative detection lnc RNA LNC_004208 expression includes following two pairs or a pair:
LNC_004208 (1)-forward primer 5 '-GCCAATCCAAACCGAAAGCC-3 '
LNC_004208 (1)-reverse primer 5 '-AGACTCAGACCCCAGGCTAC-3 '
LNC_004208 (2)-forward primer 5 '-TTTGCTACATCCCAGCTCCA-3 '
LNC_004208 (2)-reverse primer 5 '-TCCCCCTTTCCCTTGTAGGT-3 '.
Also contain reference gene β-actin specific primer in reagent:
Forward primer 5 '-CATGTACGTTGCTATCCAGGC-3 ' is shown in SEQ NO:6；
Reverse primer 5 '-CTCCTTAATGTCACGCACGAT-3 ' is shown in SEQ NO:7.
Also contain in reagent:
(1) the extracted total RNA agents useful for same from glioma cells in tissue, including stablizing solution, reagent, chloroform, isopropyl
Alcohol, without enzyme water；
It (2) is cDNA agents useful for same, including reverse transcription buffering by RNA LNC_004208 reverse transcription by template of total serum IgE
Liquid, triphosphoric acid base deoxidation nucleotide, RNase inhibitor, MMLV reverse transcriptase and random primer；
(3) by cDNA real-time quantitative PCR agents useful for same, including real time fluorescent quantitative SYBR dyestuff, without enzyme water.
The present invention is by research and utilization lncRNA sequencing discovery, the brain that lncRNA LNC_004208 is resisted in Temozolomide
Him has been inquired in detail by having carried out Effect of follow-up visit by telephone to 104 Patients with gliomas in overexpression in glioma cell line
Start time, rank, treatment condition, whether there is or not recurrence, whether there is or not suffer from again other diseases, whether there is or not use other drugs, recurrence and
Death time etc., and register life span and state.By in glioma cells in tissue LNC_004208 expression and patient
Life span and the survival analysis that carries out of state, it is found that high expression patient's mean survival time and progression free survival phase are obviously short
The patient not expressed in low expression or, and it is related with the reactivity of Temozolomide.Illustrate that LNC_004208 is pre- with glioma
Relevant molecular labeling afterwards, lncRNA LNC_004208 expression is high, patient's poor prognosis.
Detailed description of the invention
Fig. 1 is that transcript profile sequencing technologies and lncRNA sequencing technologies analyze TMZ sensitivity/resisting cell express spectra；
Fig. 2 is the lncRNA for screening glioma TMZ and resisting differential expression in cell；
Fig. 3 is the lncRNA for screening and verifying glioma TMZ and resist differential expression in cell；
Fig. 4 is the correlation that patient survival is expressed with lncRNA；
Fig. 5 is the reactivity and the correlation of lncRNA expression of patient, temozolomide.
Specific embodiment
The present invention is further illustrated below in conjunction with specific embodiment, is not intended to limit the present invention.
1 transcript profile sequencing technologies of embodiment and lncRNA sequencing technologies analyze TMZ sensitivity/resisting cell express spectra
Using transcript profile sequencing technologies and lncRNA sequencing technologies (Illumina HiSeqTM2000) glioma, is chosen
The T98G-R/U-118MG-R cell line that T98G/U-118MG cell line and TMZ are resisted carries out expression pattern analysis.It is being screened
Before, we use Cuffmerge software first, merge to the transcript that each sample is spliced, and remove wherein chain
The uncertain transcript in direction obtains the complete transcription group information of this sequencing.5 steps (Figure 1A) of screening process point or less:
Step1: transcript exon number screening: a large amount of low expression amounts in filtering transcript profile splicing result, low confidence level
Single exon transcript selects exon number > (such as without particular/special requirement, plant can retain exon number=1 to=2 transcript
Transcript)；
Step2: transcript length screening: selection transcript length > 200bp transcript；
Step3: it the known annotation screening of transcript: by Cuffcompare software, screens out and the region database annotation exon
There is the transcript of overlapping, and by database, has overlapping lncRNA as database with this splicing region transcript exon
Annotation lncRNA brings subsequent analysis into；
Step4: the screening of transcript expression quantity: calculating the expression quantity of every transcript by Cuffquant, select FPKM >
=0.5 transcript (being 2 for single exon transcript screening threshold value)；
Step5: coding potential screening: it is divided into CNCI, CPC, PFAM, phyloCSF is (when species are nonmammalian, no
Carry out phyloCSF analysis).
And (Figure 1B) is analyzed by following software:
CNCI: it is carried out by CNCI (Coding-Non-Coding Index) software.CNCI can be according to adjacent three core
The frequency spectrum of thuja acid efficiently differentiates the sequence of coding and non-coding.CNCI can effectively turn incomplete transcript and antisense
This is recorded to predicting；
CPC: it is carried out by CPC (Coding Potential Calculator, coding potential calculate) software.It will turn
Record is originally BLAST with known albumen database and is compared, and Biological Sequence feature of the CPC according to each encoder block of transcript passes through
The classifier of support vector machines assesses the coding potential of transcript.Ten folding cross validations show that CPC can be distinguished accurately
Transcript with code capacity；
PFAM: it is carried out by pfamscan software.Albumen, high quality known to Pfam-A data-base recording major part
Structural domain, and Pfam-B database then more comprehensively covers structural domain family, is the supplement to Pfam-A；We will turn
The Homology search that protein sequence and Pfam-A and Pfam-B database in this each encoder block are hmmscan is recorded, can compare and arrive
The sequence of the two databases, i.e. with the transcript of some structural domain, be considered to have code capacity, and compare less than transcription
Originally it is particularly likely that the transcript of non-coding；
PhyloCSF (carries out this analysis just for fraction of mammalian cells): being carried out by PhyloCSF software.Password
Sub- replacement frequency (Codon Substitution Freuqencies, CSF) refers to the replacement of certain codon in Multiple Sequence Alignment
Appearance frequency, coding and noncoding region codon replacement frequency ratio be one effectively distinguish one section of sequence can encode
The method of albumen；PhyloCSF can pass through in the genome alignment result of the several species of transcript in conjunction with systematic evolution tree
The codon replacement frequency of code area and noncoding region gives a mark to the coding potential of transcript.PhyloCSF utilizes several species
Between whole genome sequence compare one section of genome area of document definition whether have coding potential.Pass through literature query, Wo Menfa
PhyloCSF threshold is not quite similar between existing different species, therefore randomly chooses the known of this project research species certain amount first
Coding and lncRNA gene carries out threshold analysis, then screens candidate transcription this analysis result.This analysis is only limitted to mammal.
Final analysis result identifies 5166 lncRNA (Fig. 1 C-F) altogether.Differential expression is carried out using cuffdiff software
Analysis, screens the lncRNA (Fig. 2) of 94 differential expressions.
The lncRNA that glioma TMZ resists differential expression in cell is screened and verified to embodiment 2
(1) cell total rna extracts (TRIzol method)
By cell inoculation in 6 orifice plates, it is placed in 37 DEG C, 5%CO2It is incubated in incubator.After cell completely it is adherent cover with after,
1000 μ l TRIzol are added into every hole.It stands to after being completely dissolved, is transferred in 1.5ml EP pipe.0.2ml chloroform is added,
Acutely oscillation 15 seconds, are placed at room temperature for 3min.It is centrifuged 15min in 4 DEG C, 12000rpm, supernatant is transferred in new EP pipe.It is added
0.5ml isopropanol, is placed at room temperature for 10min.4 DEG C, 12000rpm centrifugation 10min after, there is gelatinous precipitate in EP bottom of the tube.It discards
Supernatant discards supernatant liquid with 1ml75 ℅ ethanol washing RNA precipitate, and in 4 DEG C, 7500rpm centrifugation 5min again.Air is dry
After in dry 5min, 25 μ l DEPC water are added, being beaten with pipette tips suction dissolves it sufficiently.It is placed in -80 DEG C of preservations.
(2) reverse transcription synthesizes cDNA
1. taking 5.0 μ g RNA, following reacted constituent is added in EP pipe
Mixing is jiggled after the completion of being added, is incubated at 65 DEG C 5 minutes, is then incubated for 1 minute on ice.
2. adding following substance (cDNA synthetic mixture) in order in EP pipe
It mixes gently, is collected after brief centrifugation.PCR instrument reverse transcription process: in 25 DEG C of 10min, 50 DEG C of 50min.At 85 DEG C
5min carries out termination reaction, cooled on ice.The cDNA of synthesis is placed in -20 DEG C of preservations.
(3)Real time-PCR
PCR primer sequence is as follows:
LNC_004208 (1)-forward primer 5 '-GCCAATCCAAACCGAAAGCC-3 '
LNC_004208 (1)-reverse primer 5 '-AGACTCAGACCCCAGGCTAC-3 '
LNC_004208 (2)-forward primer 5 '-TTTGCTACATCCCAGCTCCA-3 '
LNC_004208 (2)-reverse primer 5 '-TCCCCCTTTCCCTTGTAGGT-3 '.
The expression of differential expression lncRNA is further verified by Real-Time PCR, it was demonstrated that lncRNA XLOC_
024808 in the significantly high expression of T98G-R, U-118MG-R cell line, and the results are shown in attached figure 3.
The correlation that 3 patient survival of embodiment is expressed with lncRNA
By having carried out Effect of follow-up visit by telephone to 104 Patients with gliomas, inquired in detail they start time, rank,
Treatment condition, whether there is or not recurrence, whether there is or not suffer from other diseases again, whether there is or not using other drugs, recurrence and death time etc., and register
Life span and state.The life that the life span and state of LNC_004208 expression and patient in glioma cells in tissue are carried out
It deposits analysis, it is found that high expression patient's progression free survival phase (Fig. 4 A) and mean survival time are significantly shorter than low expression or do not express
Patient (Fig. 4 B).
The correlation of the reactivity of 4 patient, temozolomide of embodiment and lncRNA expression
The foundation of Human Brain Gliomas sample database
Essential information is collected: the basic conditions such as age, personal and family when the preoperative age for collecting patient, gender, morbidity
Medical history, the past treatment condition and computed tomography (CT) or Magnetic resonance imaging (MRI) result.It opens within postoperative 6 months
Begin to carry out follow-up investigation, follow-up content include include postoperative radiation and chemotherapy situation, the size of knurl and position (CT, MRI)
And the Progression free survival (Progression-free survival, PFS) of patient, total existence (Overall survival,
OS) etc..
Tumor tissues sample collection: the quasi- patients with gliomas for carrying out surgical resection therapy of selection signs informed consent form.Group
Knitting sample materials must carry out as early as possible after tissue is in vitro under aseptic condition.Obtained tissue is cut into several piece, a part mark respectively
Originally it is put into the sterile cryopreservation tube for having posted patient information, is put into portable liquid nitrogen bottle immediately, unloading enters laboratory liquid later
It is saved in nitrogen tank, the extraction for tissue RNA, DNA, total protein；Another part sample is put into 10% formaldehyde and fixes, and selects a time
Paraffin section is made in paraffin embedding.
Define TMZ sensitivity/drug resistance according to the recent evaluation criterion of WHO solid tumor: complete incidence graph (CR), part alleviate (PR),
Improvement (MR) is defined as TMZ sensitivity；Stable disease (SD), progress (PD) are defined as TMZ drug resistance.Wherein, CR refers to that tumor focus is complete
It totally disappeared mistake, and at least maintain 4 weeks or more；PR refers to volume-diminished >=50% of tumor focus, maintain 4 weeks or more；MR refers to neoplastic disease
The volume-diminished 25-50% of stove；SD refers to that the volume of tumor focus increases or reduce < 25%, and no new lesion occurs；PD refers to tumour
The volume of lesion increases >=25%, or has new lesion to occur.
The analysis of glioma cells in tissue sample
This seminar and the cooperation of Xiangya Hospital, Central-South China Univ.'s neurosurgery, collect the sample of 104 gliomas, collected
Sample is divided into TMZ sensitivity and two groups of TMZ drug resistance according to the recent evaluation criterion of WHO solid tumor.By Real-Time PCR upper
The rna expression level for detecting lncRNA LNC_004208 in glioma cells in tissue sample is stated, lncRNA LNC_004208 is analyzed
Whether there are correlations with glioma TMZ drug resistance for expression.In addition, the Clinical Study Protocol of this experiment, clinical investigation table
Lattice and patient's informed consent form are audited and are ratified by Xiangya Hospital, Central-South China Univ.'s Medical Ethics Committee.
This seminar successful collection 104 clinical case histories using the parallel resection of glioma patient with operation of TMZ chemotherapy provide
Material and follow up data, chemotherapeutic efficacy evaluation criterion is according to WHO oncotherapy reaction normals in 2000 and CA125 judgment criteria, area
It is divided into TMZ chemotherapy resistance and TMZ chemosensitivity patient.As a result, it has been found that the reactivity that lncRNA expresses high patient, temozolomide is poor
In the patient of low expression.The sensibility for the model analyzed is 56.81%, and specificity 63.63% is shown in Fig. 5.
Sequence table
<110>Xiangya Hospital, Central-South China Univ.
<120>IncRNALNC_004208 and its detection reagent are preparing the application in glioma prognostic agent
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gagaagaaat aaaagaaatc ccctctcact ctccaattct gttgttgcct ccatgcgcca 120
atccaaaccg aaagccagaa ggaagaagag tgaggagttt gctacatccc agctccacat 180
cactggaaat gggatttgag gtaatgacta ggagtttaac ctgtattccc tgcagttaga 240
aggacagtat tggtagcctg gggtctgagt ctagactaag aaatgaacat gcagcaggct 300
gtagtggttt tgtggcagca aaatcatgac ctgaagtttg gtgggaagct gttcttccac 360
agtttattgg gaaaaagact tgtaatttcc attgaccaga ccttcataga gaggcagcat 420
catctctctc ccaatccctt catccaaccg caccagctaa ttagaggacc tgattagcta 480
gaaggagaga aggagtaaaa gaattagggg aggaaaaaga ggggaagcca atcagcttcc 540
tttcttttgt tgtttgtcca cagtaggacc tacaagggaa agggggatat gcctgaaatc 600
aaggttagat tctttgatga tgacatagga ctggacattt taggggctga attgagactc 660
tagtaaatga atgaacagga attattaatt tataataaat cgtgaggcta cccagtttac 720
ctgaacaggg aaaaggggaa gaaatatctc cactcatcaa ctgtaaaaga caatgagaag 780
caaaactaga atcactttat gactacatcc catgagtcat gtttgttccc attatagtta 840
tattggcttg acctgatgat ttctgagagg aagtgttaac atcccctatt atggttatgt 900
ctttacctac tagtccttat atagttcact gggtttgctt tagcatatgt taaactgtta 960
cagctaggag aataaagact cctaactgtt ttataagttc ctagtaaaag ggactttgtg 1020
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aattatatca ctgttgccac agccattttt aaaataaatt ttactagata ttttttttcc 1140
atcctacatt tttaaacagt gtatcatttt gtttgggatg tgtctcttaa aaataccatt 1200
tagtaacttg ggttgtctat ttgttcttat tctggt 1236
<210> 2
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tttgctacat cccagctcca 20
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tccccctttc ccttgtaggt 20
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catgtacgtt gctatccagg c 21
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Claims (10)
1. a kind of IncRNALNC_004208, sequence is as shown in SEQ NO:1.
2. detecting the reagent of IncRNALNC_004208 expression quantity in preparation glioma prognosis or detection TMZ sensibility reagent
Using the sequence of the IncRNA LNC_004208 is as shown in SEQ NO:1.
3. application according to claim 2, which is characterized in that the detection IncRNA LNC_004208 expression quantity
Reagent is for detecting glioma cells in tissue.
4. application according to claim 2, which is characterized in that the detection IncRNA LNC_004208 expression quantity
Reagent includes real time fluorescent quantitative detection reagent.
5. application according to claim 4, which is characterized in that the primer of amplification lncRNA LNC_004208 includes following
It is a pair of or two pairs:
6. application according to claim 4, which is characterized in that the β-actin forward primer in detection reagent for control
For 5 '-CATGTACGTTGCTATCCAGGC-3 ', reverse primer 5 '-CTCCTTAATGTCACGCACGAT-3 '.
7. a kind of glioma prognosis or detection TMZ sensibility reagent, which is characterized in that for detection IncRNALNC_004208 table
Up to the reagent of amount, the sequence of IncRNA LNC_004208 is as shown in SEQ NO:1.
8. glioma prognosis according to claim 7 or detection TMZ sensibility reagent, which is characterized in that the inspection
The reagent for surveying IncRNA LNC_004208 expression quantity includes real time fluorescent quantitative detection reagent.
9. glioma prognosis according to claim 8 or detection TMZ sensibility reagent, which is characterized in that amplification
The primer of lncRNA LNC_004208 includes following a pair or two pairs:
LNC_004208 (1)-forward primer 5 '-GCCAATCCAAACCGAAAGCC-3 '
LNC_004208 (1)-reverse primer 5 '-AGACTCAGACCCCAGGCTAC-3 '
LNC_004208 (2)-forward primer 5 '-TTTGCTACATCCCAGCTCCA-3 '
LNC_004208 (2)-reverse primer 5 '-TCCCCCTTTCCCTTGTAGGT-3 '.
10. glioma prognosis according to claim 8 or detection TMZ sensibility reagent, which is characterized in that detection reagent
In for control β-actin forward primer be 5 '-CATGTACGTTGCTATCCAGGC-3 ', reverse primer 5 '-
CTCCTTAATGTCACGCACGAT-3’。
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110106180A (en) * | 2019-05-29 | 2019-08-09 | 中国医科大学附属盛京医院 | A kind of lncRNA molecule and its application in Treatment for Glioma/prognosis evaluation |
| CN110205320A (en) * | 2019-06-18 | 2019-09-06 | 中国医科大学附属盛京医院 | A kind of lncRNA molecule linc00998 and its application in Treatment for Glioma/prognosis evaluation |
| CN113533728A (en) * | 2021-06-03 | 2021-10-22 | 中国科学院生物物理研究所 | Biomarker of brain glioma and application thereof |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109609645B (en) * | 2019-01-22 | 2022-06-21 | 中南大学湘雅医院 | Application of reagent for detecting expression quantity of IncRNA LNC-004208 in preparation of glioma diagnostic reagent |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104587492A (en) * | 2015-01-07 | 2015-05-06 | 南方医科大学南方医院 | Application of long-chain non-coding RNA molecule SNHG18 in preparing medicine for treating brain glioma |
| CN107779508A (en) * | 2017-11-15 | 2018-03-09 | 哈尔滨医科大学 | Purposes of the lncRNA NEAT1 as molecular diagnosis mark in diagnosis of glioma reagent is prepared |
| CN108504737A (en) * | 2018-04-11 | 2018-09-07 | 中南大学湘雅医院 | Application of long-chain non-coding RNA TRA L R |
| CN109609645A (en) * | 2019-01-22 | 2019-04-12 | 中南大学湘雅医院 | The reagent of detection IncRNA LNC_004208 expression quantity is preparing the application in diagnosis of glioma reagent |
-
2019
- 2019-01-22 CN CN201910060752.XA patent/CN109679957B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104587492A (en) * | 2015-01-07 | 2015-05-06 | 南方医科大学南方医院 | Application of long-chain non-coding RNA molecule SNHG18 in preparing medicine for treating brain glioma |
| CN107779508A (en) * | 2017-11-15 | 2018-03-09 | 哈尔滨医科大学 | Purposes of the lncRNA NEAT1 as molecular diagnosis mark in diagnosis of glioma reagent is prepared |
| CN108504737A (en) * | 2018-04-11 | 2018-09-07 | 中南大学湘雅医院 | Application of long-chain non-coding RNA TRA L R |
| CN109609645A (en) * | 2019-01-22 | 2019-04-12 | 中南大学湘雅医院 | The reagent of detection IncRNA LNC_004208 expression quantity is preparing the application in diagnosis of glioma reagent |
Non-Patent Citations (8)
| Title |
|---|
| TUGCE BALCI等: "Analysis of dysregulated long non-coding RNA expressions in glioblastoma cells", 《GENE》 * |
| VECERA M 等: "Long Non-Coding RNAs in Gliomas: From Molecular Pathology to Diagnostic Biomarkers and Therapeutic Targets", 《INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES》 * |
| XU NINGBO 等: "Long noncoding RNA AC003092.1 promotes temozolomide chemosensitivity through miR-195/TFPI-2 signaling modulation in glioblastoma", 《 CELL DEATH & DISEASE》 * |
| ZENG HUIJUN 等: "Genomic profiling of long non-coding RNA and mRNA expression associated with acquiredtemozolomide resistance in glioblastoma cells", 《 INTERNATIONAL JOURNAL OF ONCOLOGY》 * |
| 徐鹏翔 等: "长链非编码RNA H19与高级别脑胶质瘤替莫唑胺抵抗的关系及其相关机制研究", 《中国临床药理学杂志》 * |
| 王向鹏: "胶质瘤相关lncRNA表达谱分析以及PAR5在胶质瘤中的功能和机制研究", 《 中国博士学位论文全文数据库 医药卫生科技辑》 * |
| 罗国轩 等: "植酸对人胶质瘤细胞U118MG增殖和凋亡的影响", 《中国医疗前沿》 * |
| 高元峰 等: "胶质瘤中异常表达的lncRNAs的研究进展", 《中国临床药理学与治疗学》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110106180A (en) * | 2019-05-29 | 2019-08-09 | 中国医科大学附属盛京医院 | A kind of lncRNA molecule and its application in Treatment for Glioma/prognosis evaluation |
| CN110106180B (en) * | 2019-05-29 | 2020-10-16 | 中国医科大学附属盛京医院 | lncRNA molecule and application thereof in glioma treatment/prognosis evaluation |
| CN110205320A (en) * | 2019-06-18 | 2019-09-06 | 中国医科大学附属盛京医院 | A kind of lncRNA molecule linc00998 and its application in Treatment for Glioma/prognosis evaluation |
| CN110205320B (en) * | 2019-06-18 | 2023-06-23 | 中国医科大学附属盛京医院 | lncRNA molecule linc00998 and application thereof in glioma treatment/prognosis evaluation |
| CN113533728A (en) * | 2021-06-03 | 2021-10-22 | 中国科学院生物物理研究所 | Biomarker of brain glioma and application thereof |
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