CN103937796A - Gastric cancer occurrence related competitive endogenous RNA (ribonucleic acid) - Google Patents
Gastric cancer occurrence related competitive endogenous RNA (ribonucleic acid) Download PDFInfo
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- CN103937796A CN103937796A CN201410150768.7A CN201410150768A CN103937796A CN 103937796 A CN103937796 A CN 103937796A CN 201410150768 A CN201410150768 A CN 201410150768A CN 103937796 A CN103937796 A CN 103937796A
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- fer1l4
- pten
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- endogenous rna
- gastric cancer
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Abstract
The invention discloses a gastric cancer occurrence related competitive endogenous RNA (ribonucleic acid). The competitive endogenous RNA is FER1L4-PTEN, wherein positive correlation property exists between FER1L4 and PTEN, namely when FER1L4 expression is high, PTEN expression is high, when the FER1L4 expression is low, the PTEN expression is low, the difference of expressions of FER1L4 in gastric cancer tissues and para-carcinoma tissues is 9.17 times, the FER1L4-PTEN is abnormal in expression in the gastric cancer tissues and has correlation with gastric cancer occurrence, and the competitive endogenous RNA has significance on further research on the effect of the gene expression regulating network in which lncRNA and mRNA take participate, in vital movement and diseases.
Description
Technical field
The present invention relates to competitive endogenous RNA, be specifically related to a pair of cancer of the stomach relevant competitive endogenous RNA occurs.
Background technology
In vivo, the transcript such as mRNA, pseudogene transcript, long-chain non-coding RNA (long noncoding RNA, lncRNA) can the identical Microrna (microRNA, miRNA) of competitive binding and the expression level that influences each other separately.These transcripts are called mutually competitive endogenous RNA (competing endogenous RNA, ceRNA).Research shows, ceRNA participates in the generation of normal activities process and disease, particularly in the generation evolution of tumour, plays an important role.
vAPA,
cNOT6L,
zEB2,
versicanwith
cD44affect the generation of the tumours such as colorectal cancer, liver cancer, mammary cancer and melanoma in ceRNA mode Deng mRNA;
pTENP1,
kRASP1with
oCT4-pg4affect the generation of the tumour such as prostate cancer and liver cancer in ceRNA mode Deng pseudogene transcript;
hULCaffect generation of liver cancer etc. Deng lncRNA in ceRNA mode.LncRNA is the desirable ceRNA of a class, because they are not translated active interference, and can be efficiently in conjunction with miRNA.Except
hULCin addition,
linc-MD1,
lincRNA-RoRwith
h19all can in ceRNA role, play a role Deng lncRNA; As the paper " Long non-coding RNA expression profile in human gastric cancer and its clinical significances " being published on " Journal of Translational Medicine " magazine for 2013 is pointed out lncRNA-
fER1L4expression level in stomach organization is lower 9.17 times than the level in cancer beside organism.
Summary of the invention
Technical problem to be solved by this invention is to provide a pair of cancer of the stomach relevant competitive endogenous RNA occurs, both
fER1L4with
pTENbetween in stomach organization, there is positive correlation, can further study gene expression regulation network that lncRNA and mRNA participate in, effect in vital movement and disease occur has great importance.
The present invention solves the problems of the technologies described above adopted technical scheme: relevant competitive endogenous RNA occurs a pair of cancer of the stomach, and this competitiveness endogenous RNA is
fER1L4-
pTEN.
The mediation miRNA of this competitiveness endogenous RNA is miR-106a-5p.
This competitiveness endogenous RNA is
fER1L4-
pTENscreening: utilize lncRNA expression level in lncRNA chip detection stomach organization and cancer beside organism, filter out the lncRNA-of 9.17 times of in stomach organization low expression
fER1L4; Use miRcode algorithm predicts with
fER1L4interactional miRNA, result shows
fER1L4on there is the binding site of miR-106a-5p; Further can adopt two luciferase report genes experimental results show that miR-106a-5p with
fER1L4interact; Obtain and experimental verification miR-106a-5p target mRNA from TarBase database, wherein
pTENfor one of target of miR-106a-5p, therefore
fER1L4with
pTENmediate by miR-106a-5p the ceRNA that partners.Adopt RT-qPCR to detect ceRNA's each other
fER1L4with
pTENexpression level in stomach organization, analyzes the dependency of its expression level, draws competitive endogenous RNA:
fER1L4with
pTENbetween in stomach organization, there is positive correlation.
Compared with prior art, the invention has the advantages that relevant competitive endogenous RNA occurs a pair of cancer of the stomach, this competitiveness endogenous RNA is
fER1L4-
pTEN;
fER1L4with
pTENbetween in stomach organization, there is positive correlation,
fER1L4express when high,
pTENexpress also high,
fER1L4express when low,
pTENexpress also low, and
fER1L4in stomach organization and cancer beside organism, express and differ 9.17 times, explanation
fER1L4-
pTENabnormal at expression in gastric cancer, there is dependency with cancer of the stomach, the effect of the gene expression regulation network participating in for further research lncRNA and mRNA in vital movement and disease generation has great importance.
Embodiment
Below in conjunction with embodiment, the present invention is described in further detail.
Use lncRNA chip Human LncRNA Array(Arraystar, Rockville, MD, USA) detect lncRNA expression level in 3 pairs of cancer of the stomach and cancer beside organism.This chip covers 18534 kinds of lncRNA.After hybridization, cleaning, use Axon GenePix 4000B Microarray Scanner(Molecular Devices, Sunnyvale, CA, USA) scan NimbleScan v2.5(Roche NimbleGen, Madison, WI, USA) analyze raw data, filter out the lncRNA-of 9.17 times of in stomach organization low expression
fER1L4.Use miRcode algorithm predicts with
fER1L4interactional miRNA, shows
fER1L4on there is the binding site of miR-106a-5p.Synthetic
fER1L4middle miR-106a-5p binding site upstream and downstream approximately 200 bp sequences, wherein in wild-type, binding site is 5 '-GCACUU-3 ', in saltant type, binding site is 5 '-UACAGG-3 ':
Wild-type sequence is:
TGAGCCTCCCCAGGCCCAGCAGGGGTCCACGTTGTCCCGGCTCACCCGAAAGAAGAAAAAGAAAGCCAGAAGGGATCAGACCCCAAAGGCGGTTCCGCAGCACTTGGACGCCAGCCCCGGTGCCGAGGGGCCTGAGATCCCCCGTGCCATGGAGGTGGAGGTGGAGGAGCTGCTGCCGCTGCCAGAGAATGTCCTGGCGCCCTGT
Saltant type sequence is:
TGAGCCTCCCCAGGCCCAGCAGGGGTCCACGTTGTCCCGGCTCACCCGAAAGAAGAAAAAGAAAGCCAGAAGGGATCAGACCCCAAAGGCGGTTCCGCATACAGGGGACGCCAGCCCCGGTGCCGAGGGGCCTGAGATCCCCCGTGCCATGGAGGTGGAGGTGGAGGAGCTGCTGCCGCTGCCAGAGAATGTCCTGGCGCCCTGT
Plasmid transfection the day before yesterday, HEKC HEK 293T is inoculated in 24 orifice plates and is cultivated.Use Lipofectamine 2000 Reagent(Invitrogen, Carlsbad, CA, USA) by triumphant miR-106a-5p eukaryon expression plasmid GV268(Ji, Shanghai, China), comprise
fER1L4the Fluc plasmid GV272(Ji of binding site sequence (wild-type or saltant type) is triumphant, Shanghai, China) and Renilla luciferase plasmid pRL-TK Vector(Promega in contrast, Madison, WI, USA) cotransfection HEK 293T cell.After transfection 24 hours, use Dual-Glo Luciferase Assay System(Promega, Madison, WI, USA) test kit processing cell, detects fluorescent value.Result demonstration, the relative fluorescence value that saltant type is a group exceeds one group approximately 56% of wild-type.This shows after binding site sudden change, miR-106a-5p couple
fER1L4restraining effect weaken, i.e. wild-type
fER1L4there is direct interaction with miR-106a-5p.TarBase database obtains the target through the miR-106a-5p of experimental verification,
pTENfor one of target of miR-106a-5p,
fER1L4with
pTENmediate by miR-106a-5p the ceRNA that partners.With in RT-qPCR detection stomach organization
fER1L4with
pTENexpression level: use GoTaq 2-Step RT-qPCR System(Promega, Madison, WI, USA) test kit carries out RT-qPCR,
fER1L4annealing temperature is 54 DEG C,
pTENannealing temperature is 53 DEG C,
gAPDHas outer reference, from 13 routine stomach organizations, analyze ceRNA's each other
fER1L4with
pTENin the dependency of expression in gastric carcinoma level, obtain table 1 result.The primer sequence is as follows respectively:
fER1L4forward primer: 5 '-CCGTGTTGAGGTGCTGTTC-3 '
fER1L4reverse primer: 5 '-GGCAAGTCCACTGTCAGATG-3 '
pTENforward primer: 5 '-GTTTACCGGCAGCATCAAAT-3 '
pTENreverse primer: 5 '-CCCCCACTTTAGTGCACAGT-3 '
gAPDHforward primer: 5 '-AAGGTGAAGGTCGGAGTCAA-3 '
gAPDHreverse primer: 5 '-AATGAAGGGGTCATTGATGG-3 '
In table 1 stomach organization
fER1L4with
pTENrelative expression's level
Stomach organization numbering | FER1L4Relative expression's level | PTENRelative expression's level |
1 | 0.23 | 1.27 |
2 | 0.19 | 0.74 |
3 | 0.28 | 1.10 |
4 | 0.21 | 0.90 |
5 | 0.26 | 1.11 |
6 | 0.18 | 0.93 |
7 | 0.29 | 1.08 |
8 | 0.25 | 0.93 |
9 | 0.26 | 0.94 |
10 | 0.37 | 2.2 |
11 | 0.22 | 0.95 |
12 | 0.20 | 0.62 |
13 | 0.29 | 0.87 |
From table 1, result shows:
fER1L4with
pTENexpression level in stomach organization is proportionate, relation conefficient
rbe 0.77.The gene expression regulation network that so further research lncRNA and mRNA participate in, the effect in vital movement and disease generation have great importance.
<110> University Of Ningbo
There is relevant competitive endogenous RNA in a pair of cancer of the stomach of <120>
<160> 8
<170> PatentIn version 3.5
<210> 1
<211> 205
<212> RNA
<400> 1
TGAGCCTCCC CAGGCCCAGC AGGGGTCCAC GTTGTCCCGG CTCACCCGAA50 AGAAGAAAAA GAAAGCCAGA AGGGATCAGA CCCCAAAGGC GGTTCCGCAG100 CACTTGGACG CCAGCCCCGG TGCCGAGGGG CCTGAGATCC CCCGTGCCAT150 GGAGGTGGAG GTGGAGGAGC TGCTGCCGCT GCCAGAGAAT GTCCTGGCGC200 CCTGT 205
<210> 2
<211> 205
<212> RNA
<400> 2
TGAGCCTCCC CAGGCCCAGC AGGGGTCCAC GTTGTCCCGG CTCACCCGAA50 AGAAGAAAAA GAAAGCCAGA AGGGATCAGA CCCCAAAGGC GGTTCCGCAT100 ACAGGGGACG CCAGCCCCGG TGCCGAGGGG CCTGAGATCC CCCGTGCCAT150 GGAGGTGGAG GTGGAGGAGC TGCTGCCGCT GCCAGAGAAT GTCCTGGCGC200 CCTGT 205
<210>3
<211> 19
<212> DNA
<213> artificial sequence
<400> 3
CCGTGTTGAG GTGCTGTTC 19
<210> 4
<211> 20
<212> DNA
<213> artificial sequence
<400> 4
GGCAAGTCCA CTGTCAGATG 20
<210> 5
<211> 20
<212> DNA
<213> artificial sequence
<400> 5
GTTTACCGGC AGCATCAAAT 20
<210> 6
<211> 20
<212> DNA
<213> artificial sequence
<400> 6
CCCCCACTTT AGTGCACAGT 20
<210> 7
<211> 20
<212> DNA
<213> artificial sequence
<400> 7
AAGGTGAAGG TCGGAGTCAA 20
<210> 8
<211> 20
<212> DNA
<213> artificial sequence
<400> 8
AATGAAGGGG TCATTGATGG 20
Claims (2)
1. there is relevant competitive endogenous RNA in a pair of cancer of the stomach, it is characterized in that this competitiveness endogenous RNA is
fER1L4-
pTEN.
2. there is relevant competitive endogenous RNA in a pair of cancer of the stomach as claimed in claim 1, and the mediation miRNA that it is characterized in that this competitiveness endogenous RNA is miR-106a-5p.
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Cited By (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104651364A (en) * | 2015-02-04 | 2015-05-27 | 中国人民解放军第二军医大学 | LncRNA capable of competitively consuming carcinogenic microRNAs, oncolytic adenovirus and application thereof |
CN106202993A (en) * | 2016-07-12 | 2016-12-07 | 王亚帝 | Utilize the method that mrna expression spectrum combines screening cardiotoxicity induced by anthracyclines gene with competitive endogenous RNA express spectra |
CN109576373A (en) * | 2019-01-02 | 2019-04-05 | 首都医科大学附属北京朝阳医院 | Application of the circ-VAPA as gastric cancer and diagnosis of colorectal carcinoma biomarker and therapy target |
CN111471772A (en) * | 2019-06-06 | 2020-07-31 | 徐州医科大学 | Molecular marker for cancer diagnosis |
CN113005179A (en) * | 2021-02-26 | 2021-06-22 | 广东省中医院(广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院) | Mass spectrum method for quantifying nucleic acid based on DNA-polypeptide probe technology and application |
-
2014
- 2014-04-16 CN CN201410150768.7A patent/CN103937796A/en active Pending
Non-Patent Citations (4)
Title |
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MEMORIAL SLOAN-KETTERING CANCER CENTER: "Pten target,FER1L4 target", 《MICRORNA.ORG - TARGETS AND EXPRESSION》 * |
SONG ET AL: "Long non-coding RNA expression profile in human gastric cancer and its clinical significances", 《JOURNAL OF TRANSLATIONAL MEDICINE》 * |
夏天等: "竞争性内源RNA: 一种全新的基因表达调控模式", 《中国生物化学与分子生物学报》 * |
王茂林等: "PTEN 基因在胃癌组织中的表达及临床意义", 《疾病监测与控制杂志》 * |
Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104651364A (en) * | 2015-02-04 | 2015-05-27 | 中国人民解放军第二军医大学 | LncRNA capable of competitively consuming carcinogenic microRNAs, oncolytic adenovirus and application thereof |
CN104651364B (en) * | 2015-02-04 | 2017-07-07 | 中国人民解放军第二军医大学 | A kind of LncRNA, oncolytic adenovirus of competitive consumption carcinogenicity microRNAs and application thereof |
CN106202993A (en) * | 2016-07-12 | 2016-12-07 | 王亚帝 | Utilize the method that mrna expression spectrum combines screening cardiotoxicity induced by anthracyclines gene with competitive endogenous RNA express spectra |
CN106202993B (en) * | 2016-07-12 | 2018-09-25 | 王亚帝 | A method of joint screening cardiotoxicity induced by anthracyclines gene |
CN109576373A (en) * | 2019-01-02 | 2019-04-05 | 首都医科大学附属北京朝阳医院 | Application of the circ-VAPA as gastric cancer and diagnosis of colorectal carcinoma biomarker and therapy target |
CN111471772A (en) * | 2019-06-06 | 2020-07-31 | 徐州医科大学 | Molecular marker for cancer diagnosis |
CN113005179A (en) * | 2021-02-26 | 2021-06-22 | 广东省中医院(广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院) | Mass spectrum method for quantifying nucleic acid based on DNA-polypeptide probe technology and application |
CN113005179B (en) * | 2021-02-26 | 2021-11-16 | 广东省中医院(广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院) | Mass spectrum method for quantifying nucleic acid based on DNA-polypeptide probe technology and application |
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