CN105506158B - The application method of long-chain non-coding RNA LOC284454 - Google Patents
The application method of long-chain non-coding RNA LOC284454 Download PDFInfo
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- CN105506158B CN105506158B CN201610067499.7A CN201610067499A CN105506158B CN 105506158 B CN105506158 B CN 105506158B CN 201610067499 A CN201610067499 A CN 201610067499A CN 105506158 B CN105506158 B CN 105506158B
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Abstract
The invention discloses the application of the reagent of long-chain non-coding RNA LOC284454 expression quantity in detection ovarian cancer tissue, it is mainly used for preparing the real time fluorescent quantitative detection reagent of oophoroma auxiliary diagnosis or outcome prediction.Confirm that LOC284454 expresses up-regulation in ovarian cancer tissue by research, ovarian cancer patients poor prognosis of the ovarian cancer patients than low expression LOC284454 of height expression LOC284454, therefore, the expression of LOC284454 is used for the prognosis prediction of ovarian cancer patients, strong molecular biology foundation can be provided for the prognosis of ovarian cancer patients, there is far-reaching clinical meaning and important popularization and application foreground.
Description
Technical field
The invention belongs to oncomolecularbiology fields, and in particular to long-chain non-coding RNA in detection ovarian cancer tissue
The reagent of LOC284454 expression quantity is used to prepare the purposes of oophoroma auxiliary 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, that is, malignant tumor of ovary is common one of the malignant tumour of female sex organ, and disease incidence is only second to son
Cervical carcinoma and carcinoma of uterine body and arrange and occupy third position.But ovarian epithelium mortality of carcinoma accounts for the first place of all kinds of gynecological tumors, to women
Life causes to seriously threaten.Since the embryonic development of ovary, anatomic tissue and endocrine function are more complex, 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 LOC284454 (NCBI accession number:NR_036515) in oophoroma in oophoroma
The significant up-regulation of expression, LOC284454 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 reagents of long-chain non-coding RNA LOC284454 expression quantity in detection ovarian cancer tissue
It is used to prepare the purposes of oophoroma auxiliary diagnosis or outcome prediction preparation.
Detect the application of the reagent of long-chain non-coding RNA LOC284454 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 LOC284454
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 LOC284454 expression in the real time fluorescent quantitative detection reagent
Object sequence:
Forward primer: 5 '-TGAAATCAGATGGGAGGAGGC-3 ',
Reverse primer: 5 '-AGACATGACACACAGCACGA-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 LOC284454, LOC284454 expresses up-regulation in ovarian cancer tissue as the result is shown.Prompt LOC284454 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 that LOC284454 expresses up-regulation situation in ovarian cancer tissue；
Real-Time Fluorescent Quantitative PCR Technique detects lncRNA LOC284454 by oophoroma (28) and oophoroma (12)
In expression, P < 0.001.
Fig. 2 is the relationship of expression Yu the ovarian cancer patients prognosis of LOC284454 in oophoroma；
The high expression of LOC284454 is related to the poor prognosis of ovarian cancer patients in oophoroma, i.e. LOC284454 high expression
The total life span (Over-all survival) of patient will significantly lower than LOC284454 low expression or the patient not expressed,
LOC284454 expresses high patient and is easier to recur.
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 LOC284454 express in oophoroma up-regulation 1. materials and
Method:
Collect by 12 oophoromas and 28 ovarian cancer tissues, extracted total RNA, 1 μ g RNA after reverse transcription is at cDNA, into
Row real-time fluorescence quantitative PCR.LOC284454 forward primer is 5 '-
TGAAATCAGATGGGAGGAGGC-3 ', as shown in SEQ NO:2 and reverse primer 5 '-
AGACATGACACACAGCACGA-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
1 94℃ 5min
2 95℃ 10sec
3 58℃ 30sec
4 72℃ 20sec
5 Plate read
6 82℃ 30sec
7 Plate read
8 Go to step 2for more 39times
9 55.0 DEG C of from of Perform melting curve 95.0 DEG C of to, 0.2 DEG C of read every, hold
for 1sec
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
LOC284454 do not express or express in control tissue by the cancer it is very low, and in ovarian cancer tissue high expression P <
0.001 (Fig. 1)
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 LOC284454 and the life span of patient and state carry out, discovery LOC284454 high express patient
Mean survival time is significantly shorter than LOC284454 low expression or the patient not expressed (Fig. 2).Illustrate LOC284454 be one with
The relevant molecular labeling of ovarian cancer prognosis, lncRNA expression is high, patient's poor prognosis.
Claims (4)
1. detecting the application of the reagent of long-chain non-coding RNA LOC284454 expression quantity in ovarian cancer tissue, which is characterized in that institute
The reagent stated is used to prepare oophoroma outcome prediction preparation；The sequence such as SEQ NO:1 of long-chain non-coding RNA LOC284454
It is shown.
2. application according to claim 1, which is characterized in that the oophoroma outcome prediction preparation is fixed for real-time fluorescence
Measure 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 LOC284454 expression:
Forward primer: 5 '-TGAAATCAGATGGGAGGAGGC-3 ',
Reverse primer: 5 '-AGACATGACACACAGCACGA -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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Families Citing this family (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105907858A (en) * | 2016-05-04 | 2016-08-31 | 李青 | Method of enabling lncRNA RP11 697E22.2 to be used as reversal ovarian carcinoma intervention target |
| 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 |
| CN115232874B (en) * | 2021-09-24 | 2023-04-07 | 四川大学华西第二医院 | Application of long-chain non-coding RNA in regulation and control of ovarian cancer progression |
Citations (2)
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|---|---|---|---|---|
| CN102433326A (en) * | 2011-04-25 | 2012-05-02 | 北京大学 | Long non-coding RNA and application thereof |
| CN104388556A (en) * | 2014-11-11 | 2015-03-04 | 中国人民解放军第二军医大学 | Application of long non-coding RNA in preparation of medicament for diagnosing or treating ovarian cancer |
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| US9410206B2 (en) * | 2011-11-30 | 2016-08-09 | John Wayne Cancer Institute | Long noncoding RNA (lncRNA) as a biomarker and therapeutic marker in cancer |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102433326A (en) * | 2011-04-25 | 2012-05-02 | 北京大学 | Long non-coding RNA and application thereof |
| CN104388556A (en) * | 2014-11-11 | 2015-03-04 | 中国人民解放军第二军医大学 | Application of long non-coding RNA in preparation of medicament for diagnosing or treating ovarian cancer |
Non-Patent Citations (2)
| Title |
|---|
| Histo-genomic stratification reveals the frequent amplification/overexpression of CCNE1 and BRD4 genes in non-BRCAness high grade ovarian carcinoma;Oumou Goundiam等;《International Journal of Cancer》;20150421;第137卷;第1896页左栏第2段至右栏第1 段 * |
| NCBI Reference Sequence: NR_036515.1;无;《GenBank》;20150521;核酸序列 * |
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