CN106048073B - Biomarker and its application of a kind of oophoroma auxiliary diagnosis or outcome prediction - Google Patents
Biomarker and its application of a kind of oophoroma auxiliary diagnosis or outcome prediction Download PDFInfo
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- CN106048073B CN106048073B CN201610688244.2A CN201610688244A CN106048073B CN 106048073 B CN106048073 B CN 106048073B CN 201610688244 A CN201610688244 A CN 201610688244A CN 106048073 B CN106048073 B CN 106048073B
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Abstract
The invention discloses a kind of oophoroma auxiliary diagnosis, perhaps the biomarker of outcome prediction and its application are mainly used for preparing the real time fluorescent quantitative detection reagent of oophoroma auxiliary diagnosis or outcome prediction.Confirm that LINC01420 expresses up-regulation in ovarian cancer tissue by research, ovarian cancer patients poor prognosis of the ovarian cancer patients of highly expressed LINC01420 than low expression LINC01420, therefore, the expression of LINC01420 can provide strong molecular biology foundation for the prognosis of ovarian cancer patients, have far-reaching clinical meaning and important popularization and application foreground.
Description
Technical field
The invention belongs to oncomolecularbiology fields, and in particular to a kind of oophoroma auxiliary diagnosis or outcome prediction are used
Biomarker long-chain non-coding RNA LINC01420, and detect the reagent of the rna expression amount to be used to prepare oophoroma auxiliary
Help the purposes of diagnosis or outcome prediction preparation.
Background technique
The Human Genome Project and its subsequent DNA element encyclopedia plan (The Encyclopedia of DNA
Elements Project, ENCODE) research achievement shows that protein coding gene sequence only accounts for the 1- of human genomic sequence
3%, and in human genome the transcribed sequences of the overwhelming majority be long-chain non-coding RNA (Long non-coding RNA,
lncRNA).LncRNA is universally present in various biologies, and with the raising of biological complex degree, lncRNA in genome
The ratio of sequence also correspondingly increases, and prompts lncRNA significant during biological evolution.With lncRNA constantly quilt
It was found that their function is gradually interpreted, scientists discovery lncRNA actively participates in vital movement different layers extensively
In the function controlling in face, the field brand-new as one has become international life science field new forward position and hot spot.
LncRNA can be used as signal (signal), induction (guide), bait (decoy) or the bracket of functional protein
(scaffold) various ways such as molecule regulate and control base in chromatin reconstruct, genetic transcription, translation and the multiple levels such as protein modified
The expression of cause, and irreplaceable role is played during including the basic physiologicals such as development, immune, reproduction, it is prior
It is that the expression of lncRNA and functional disturbance are closely connected with a variety of diseases of the mankind including malignant tumour one
It rises.Therefore, the function of lncRNA is furtherd investigate, is disclosed by the lncRNA hereditary information transfer mode mediated and expression regulation net
Network not only can annotate and illustrate again the structure and function of genome from the angle other than protein coding gene, deeply find
The essence and rule of vital movement are expected to recognize from a new visual angle a variety of mankind's common diseases including tumour
Pathogenesis, and new molecular marker and therapy target are provided for the Clinics and Practices of these diseases.
Oophoroma is common one of the malignant tumour of female reproductive system, and the death rate occupies first of gynecological tumor, to women
Life causes to seriously threaten.Since the tissue anatomical structure and endocrine function of ovary are more complex, oophoroma early symptom not allusion quotation
Type, the organization type of preoperative identification of ovarian tumour and good pernicious extremely difficult.The occurrence and development of tumour are polygenes participation, multistep
Suddenly, multistage complex process, LncRNA may also play important role during the occurrence and development of oophoroma.Recently
We have found lncRNA LINC01420 (NCBI accession number:NR_015367) in oophoroma in oophoroma
The significant up-regulation of expression, LINC01420 express high patient and are easier to shift, and it is low that life span is shorter than lncRNA expression
Patient, therefore can be used for the Index for diagnosis of oophoroma for the detection preparation of the lncRNA.
Summary of the invention
The object of the present invention is to provide the biomarker long-chain of a kind of oophoroma auxiliary diagnosis or outcome prediction is non-
The reagent of the rna expression amount is used to prepare oophoroma auxiliary diagnosis in coding RNA LINC01420, and detection ovarian cancer tissue
Or the purposes of outcome prediction preparation.
The biomarker of a kind of oophoroma auxiliary diagnosis or outcome prediction is long-chain non-coding RNA
LINC01420, the sequence of long-chain non-coding RNA LINC01420 is as shown in SEQ NO:1.
Detect the application of the reagent of long-chain non-coding RNA LINC01420 expression quantity in ovarian cancer tissue, the reagent
It is used to prepare oophoroma auxiliary diagnosis or outcome prediction preparation；The sequence such as SEQ of long-chain non-coding RNA LINC01420
Shown in NO:1.
The oophoroma auxiliary diagnosis or outcome prediction preparation is real time fluorescent quantitative detection reagent.
Include drawing for real time fluorescent quantitative detection LINC01420 expression in the real time fluorescent quantitative detection reagent
Object sequence:
Forward primer: 5'-CACTCTACCCTCCGCACC-3',
Reverse primer: 5'-AGGAAGTGAAATCGTGCTGA-3'.
Include reference gene GAPDH Specific PCR primers in the real time fluorescent quantitative detection reagent:
Forward primer 5 '-ACCACAGTCCATGCCATCAC-3 '
Reverse primer 5 '-TCCACCACCCTGTTGCTGTA-3 '.
The present invention is from reverse transcription, real time fluorescent quantitative method have detected after extract RNA in sample by oophoroma and oophoroma
The expression of LINC01420, LINC01420 expresses up-regulation in ovarian cancer tissue as the result is shown.Prompt LINC01420 can be used as ovum
The molecular marker of nest cancer prognosis prediction.The present invention provides strong molecule life for the auxiliary diagnosis and prognosis prediction of oophoroma
Object tool has far-reaching clinical meaning and important popularization and application foreground.
Detailed description of the invention
Fig. 1 is the relationship of expression Yu the ovarian cancer patients prognosis of LINC01420 in oophoroma；
Real-Time Fluorescent Quantitative PCR Technique detects expression of the lncRNA LINC01420 in oophoroma (28), ovum
In nest cancer the high expression of LINC01420 is related to the poor prognosis of ovarian cancer patients, i.e. the highly expressed patient of LINC01420 is always
Life span (Over-all survival) will be significantly lower than LINC01420 low expression or the patient not expressed, LINC01420
High patient is expressed to be easier due to tumor recurrence, transfer and death.
Specific embodiment
The present invention is further illustrated below in conjunction with specific embodiment, is not intended to limit the present invention.
Embodiment 1, quantitative real-time PCR detection confirm that LINC01420 expresses up-regulation in oophoroma
1. materials and methods:
28 ovarian cancer tissues, extracted total RNA are collected, 1 μ g RNA carries out real time fluorescent quantitative after reverse transcription is at cDNA
PCR.5 '-CACTCTACCCTCCGCACC-3 ' of LINC01420 forward primer, as shown in SEQ NO:2 and reverse primer 5 '-
AGGAAGTGAAATCGTGCTGA-3'.As shown in SEQ NO:3.
GAPDH forward primer for control is 5 '-ACCACAGTCCATGCCATCAC-3 ' as shown in SEQ NO:4, and
Reverse primer 5 '-TCCACCACCCTGTTGCTGTA-3 ', as shown in SEQ NO:5.
Real-time fluorescence quantitative PCR reaction system
Real-time fluorescence quantitative PCR reaction step
The amplification curve and melting curve of real-time fluorescence quantitative PCR, the expression intensity root of each gene are confirmed after reaction
After CT value (threshold cycle values), reference gene (GAPDH) markization, using group t-test checking computation
P value.
2. result
Effect of follow-up visit by telephone has been carried out to 28 ovarian cancer patients, inquired in detail they start time, treatment condition, whether there is or not
Recurrence, whether there is or not suffering from other diseases, recurrence and death time etc. again, and register life span and state, and to ovarian cancer tissue
The survival analysis that the expression of middle LINC01420 and the life span of patient and state carry out, discovery LINC01420 high express patient
(High, the 16) mean survival time is significantly shorter than LINC01420 low expression or the patient not expressed (Low, 12, Fig. 1).It says
Bright LINC01420 is a molecular labeling relevant to ovarian cancer prognosis, and lncRNA expression is high, patient's poor prognosis.
Claims (4)
1. detecting the application of the reagent of long-chain non-coding RNA LINC01420 expression quantity in ovarian cancer tissue, which is characterized in that institute
The reagent stated is used to prepare oophoroma auxiliary diagnosis or outcome prediction preparation；The sequence of long-chain non-coding RNA LINC01420
As shown in SEQ NO:1.
2. application according to claim 1, which is characterized in that the oophoroma auxiliary diagnosis or outcome prediction preparation
For real time fluorescent quantitative detection reagent.
3. application according to claim 2, which is characterized in that comprising being used in the real time fluorescent quantitative detection reagent
Real time fluorescent quantitative detects the primer sequence of LINC01420 expression:
Forward primer: 5 '-CACTCTACCCTCCGCACC-3 ',
Reverse primer: 5 '-AGGAAGTGAAATCGTGCTGA-3 '.
4. application according to claim 2 or 3, which is characterized in that include in the real time fluorescent quantitative detection reagent
Reference gene GAPDH Specific PCR primers:
Forward primer 5 '-ACCACAGTCCATGCCATCAC-3 '
Reverse primer 5 '-TCCACCACCCTGTTGCTGTA-3 '.
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| CN110066872A (en) * | 2019-03-01 | 2019-07-30 | 浙江理工大学 | LncRNA UCA1 is as the application in ovarian cancer diagnosis or the biomarker of outcome inspection |
| CN112980960B (en) * | 2021-04-23 | 2022-05-10 | 中南大学 | Application method of long-chain non-coding RNA LOC730101 and detection and treatment preparation |
| CN113234819B (en) * | 2021-04-23 | 2022-03-18 | 中南大学 | Application method of long-chain non-coding RNA LOC107987064 and detection and treatment preparation |
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| CN104388543A (en) * | 2014-10-27 | 2015-03-04 | 中南大学 | In situ hybridization probe, reagent and application of long non-coding RNA LOC401317 |
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| CN104388543A (en) * | 2014-10-27 | 2015-03-04 | 中南大学 | In situ hybridization probe, reagent and application of long non-coding RNA LOC401317 |
Non-Patent Citations (3)
| Title |
|---|
| BC062451.1;Anonymous;《GenBank》;20050509;第1-2页 |
| Long non-coding RNA expression in primary human monocytes;Hoda Mirsafian et al;《Genomics》;20160108;第108卷(第1期);第37-45页 |
| 长链非编码RNA AFAP1-AS1在消化系统肿瘤中的表达及意义;赵艳华等;《肿瘤》;20140131;第34卷(第1期);第39-46页 |
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