CN108478797A - ATF3 is preparing the application in treating nasopharyngeal carcinoma Paclitaxel Chemotherapy hypersitization medicine as target site - Google Patents
ATF3 is preparing the application in treating nasopharyngeal carcinoma Paclitaxel Chemotherapy hypersitization medicine as target site Download PDFInfo
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Abstract
Application the invention discloses ATF3 as target site in preparing treatment nasopharyngeal carcinoma Paclitaxel Chemotherapy hypersitization medicine.The present invention's research shows that activating transcription factor 3 (3 ATF3 of Activating transfactor) expression in nasopharyngeal carcinoma taxol resistance cell is significantly lower than paclitaxel-sensitive cell, and ATF3 can increase the chemosensitivity of pacilitaxel of nasopharyngeal carcinoma by regulating and controlling HIPPO signal paths.Therefore the potential target spot that ATF3 may be treated as nasopharyngeal carcinoma chemotherapy enhanced sensitivity.Therefore, ATF3 can be used for preparing the chemotherapy sensitizing medicine of nasopharyngeal carcinoma as target site.Especially ATF3 can be used for preparing the drug that regulation and control HIPPO accesses increase nasopharyngeal carcinoma cell to chemosensitivity of pacilitaxel as target site.The NPC is acute pyogenic infection of finger tip nasopharyngeal carcinoma.
Description
Technical field
The invention belongs to biomedicine technical fields, and in particular to ATF3 is preparing treatment nasopharyngeal carcinoma taxol as target spot
Application in chemical therapy sensitivity-enhancing.
Background technology
Nasopharyngeal carcinoma is a kind of squamous cell carcinoma for being common in East Asia, Southeast Asia and North African Area Highly invasive, except heredity
Except factor and environmental factor, Epstein-Barr virus plays important role in its pathogenesis, this is also that it is different from other
In place of head and neck neoplasm, nasopharyngeal carcinoma cell is relatively high for the sensibility of radioactive ray and the anatomical structure of incidence is extremely multiple
It is miscellaneous, therefore for the Nasopharyngeal Carcinoma Patients in early stage, radiotherapy is extremely effective preferred therapeutic modality.AJCC according to latest edition
(American Joint Commission on Cancer AJCC) Staging System has about 60%~70% nasopharyngeal carcinoma to suffer from
Person has been in III phases or IV phases when making a definite diagnosis, and simple is poor for the Nasopharyngeal Carcinoma Patients prognosis of late stage using radiotherapy, with
A series of researchs and clinical data afterwards show putting complex treatment and first use new adjuvant chemotherapy effective before radiotherapy
Local-advanced nasopharyngeal carcinoma survival is improved, and taxol is then a line medicine of new adjuvant chemotherapy before Synchronous chemoradiotherapy and radiotherapy
Object, but treatment failure is led to due to acquired resistance with the progressive part patient for the treatment of, explore nasopharyngeal carcinoma taxol
Drug resistant molecular mechanism and the method for finding taxol enhanced sensitivity are particularly significant and intentional for the treatment of clinical superior nasopharynx
Justice.
Long-chain non-coding RNA (long non coding RNA lncRNA) is that a kind of length is more than 200 nucleotide
Albumen is not encoded or can encode the transcription product of a small amount of small peptide section.More and more evidences show that lncRNA can pass through
A series of mechanism such as the regulation and control after the epigenetics of DNA is modified or chromatin level modulation is transcribed and transcription participate in cells and increase
Grow the pathological processes such as apoptosis, autophagy, invasion transfer and radiation resistance.It is more to find that lncRNA may participate in spite of part research
Medicine drug resistance is adjusted, but lncRNA adjust nasopharyngeal carcinoma chemosensitivity of pacilitaxel research it is very rare and specific mechanism is still unknown
Really.Therefore, finding the relevant lncRNA of nasopharyngeal carcinoma chemosensitivity of pacilitaxel has highly important clinical meaning.
Invention content
This research is intended to provide a kind of novel targets can be used for treating nasopharyngeal carcinoma Paclitaxel Chemotherapy enhanced sensitivity.In the present invention
In, we carry out the strain of nasopharyngeal carcinoma cell CNE-1 taxol resistances, the strain of HNE-2 taxol resistances and its corresponding parent plant
LncRNA chips test and analyze and are found that MRVI1-AS1 can form positive feedback signal ring with transcription factor ATF3 and pass through regulation and control
HIPPO signal paths increase nasopharyngeal carcinoma chemosensitivity of pacilitaxel.Therefore the discovery in the present invention very likely becomes nasopharynx
The new method of cancer taxol enhanced sensitivity.
We carry out chip results first to the strain of nasopharyngeal carcinoma taxol resistance and parent's plant lncRNA cDNA microarrays
Analysis finds that expressions of the MRVI1-AS1 in persister is significantly lower than parent plant, and subsequent qRT-PCR confirms MRVI1-AS1
The low expression in nasopharyngeal carcinoma taxol resistance strain, the function of further cell experiment verification MRVI1-AS1, the results show that
MRVI1-AS1 is overexpressed in persister can dramatically increase nasopharyngeal carcinoma chemosensitivity of pacilitaxel, we are into one in zoopery
The function of confirming MRVI1-AS1 of step.
This research is it has also been found that ATF3 expressions in nasopharyngeal carcinoma persister are less than parent plant, and MRVI1-AS1 can lead to
Crossing ceRNA mechanism, targeted inhibition miR-513a-5p and miR-27b-3p raises the expression of ATF3 simultaneously, further
Research finds that the expression that ATF3 is raised in nasopharyngeal carcinoma can also increase nasopharyngeal carcinoma chemosensitivity of pacilitaxel.
In the next research, we have further found that ATF3 can be opened as transcription factor with MRVI1-AS1
Sub-area combines to promote the expression of MRVI1-AS1, this shows that MRVI1-AS1 can form one mutually just with ATF3
The regulation and control ring of feedback.And MRVI1-AS1/ATF3 regulation and control rings can adjust the activity of HIPPO accesses, finally we have found that passing through tune
Section HIPPO accesses can increase nasopharyngeal carcinoma chemosensitivity of pacilitaxel.
In short, present invention demonstrates that expression is less than parent plant in ATF3 nasopharyngeal carcinoma persisters, being overexpressed ATF3 can pass through
Regulate and control HIPPO signal paths to increase nasopharyngeal carcinoma chemosensitivity of pacilitaxel.Therefore, ATF3 may be used as nasopharyngeal carcinoma chemotherapy enhanced sensitivity
Potential target spot.
Therefore, ATF3 is preparing the application in treating nasopharyngeal carcinoma Paclitaxel Chemotherapy hypersitization medicine as target site, especially
ATF3 is being prepared as target site by regulating and controlling HIPPO accesses increase nasopharyngeal carcinoma cell in the drug of chemosensitivity of pacilitaxel
Application.The nasopharyngeal carcinoma refers to NPC.
Description of the drawings
Fig. 1：A：Mtt assay detects the proliferation after two pairs of nasopharyngeal carcinoma cell taxol treatments, the proliferation of HNE-2/T, CNE-1/T
It is apparently higher than CNE-1 and HNE-2.B：Chip results prompt fold differences ＞ 5, and the lncRNA of length ＜ 2000 shares 6.C：
It is transfected in CNE-1/T after MRVI1-AS1 is overexpressed plasmid and uses taxol treatment, mtt assay is as a result, it has been found that MRVI1-AS1 can increase
CNE-1/T cell paclitaxel-sensitives, TAXOL are CNE-1/T cell IC50 concentration.D-F：Flow cytometry, Clone formation are real
It tests and scratch experiment is further discovered that CNE-1/T cell paclitaxel-sensitives can be increased by being overexpressed MRVI1-AS1 in CNE-1/T
Property, TAXOL is CNE-1/T cell IC25 concentration.G-H：Transplantable tumor experiment finds to use purple after CNE-1/T is overexpressed MRVI1-AS1
China fir alcohol is significantly greater than control group to the inhibiting effect of tumor volume, and TAXOL is CNE-1/T cell IC25 concentration.I：Transplantable tumor is exempted from
Epidemic disease groupization finds that CNE-1/T is overexpressed MRVI1-AS1 and with after taxol treatment, the expression of ki67 is decreased obviously, and TAXOL is
CNE-1/T cell IC25 concentration.Each group histogram data is the average value of independent repeated trials three times；Bars indicates standard
Difference, * p ＜ 0.05, * * p ＜ 0.01.
Fig. 2：A：Fluorescence in situ hybridization result shows that MRVI1-AS1 is expressed in simultaneously in the core of CNE-1 cells and in cytoplasm.
B-C：The prompt of miRNA chips is overexpressed miR-513a-5p and miR-27b-3p expression quantity when CNE-1/T is overexpressed MRVI1-AS1
Significantly lower than CNE-1/T control groups, and there may be same with miR-513a-5p and miR-27b-3p simultaneously by MRVI1-AS1
Binding site.D:QRT-PCR shows that miR-513a-5p and miR-27b-3p expression quantity is apparently higher than CNE-1 in CNE-1/T, and
Their expression is decreased obviously after being overexpressed MRVI1-AS1.E-F：Luciferase reporter gene and RNA pull down
Experimental result shows that MRVI1-AS1 can be directly targeted miR-513a-5p and miR-27b-3p.Each group histogram data is three times
The average value of independent repeated trials；Bars indicates that standard deviation, * p ＜ 0.05, * * p ＜ 0.01, * * * p ＜ 0.001, TAXOL represent
CNE-1/T cell IC25 concentration taxols.
Fig. 3：A：MTT experiment result indicates that the expression for inhibiting miR-513a-5p and miR-27b-3p can increase Japanese yew
Lethality of the alcohol to nasopharyngeal carcinoma cell.B-D：Flow cytometry, colony formation, scratch experiment result are shown in CNE-1/T
Middle inhibition miR-513a-5p and miR-27b-3p and with after taxol treatment, cell Proliferation and transfer are considerably less than control group, carefully
Born of the same parents' apoptosis is then higher than control group.Each group histogram data is the average value of independent repeated trials three times；Bars indicates standard
Difference, * p ＜ 0.05, * * p ＜ 0.01, * * * p ＜ 0.001, TAXOL represent CNE-1/T cell IC25 concentration taxols
Fig. 4：A-B：Chip results show the expression quantity of ATF3 when CNE-1/T is overexpressed MRVI1-AS1 compared to CNE-1/T
Rising occurs in group, and miR-513a-5p, miR-27b-3p and ATF3 have same binding site.C-F：WB and qRT-PCR is aobvious
Show that MRVI1-AS1 can express water by MRVI1-AS1 by inhibiting miR-513a-5p and miR-27b-3p to restore ATF3.G：
Dual-Luciferase experiment shows that miR-513a-5p and miR-27b-3p can target ATF3.After H is overexpressed ATF3 in CNE-1/T
With taxol treatment, MTT detections find that ATF3 can increase CNE-1/T cell paclitaxel-sensitives.Each group histogram data is
The average value of independent repeated trials three times；Bars indicates standard deviation, * p ＜ 0.05, * * p ＜ 0.01, * * * p ＜ 0.001, TAXOL
Concentration for the treatment of is CNE-1/T IC25 paclitaxel concentrations
Fig. 5：A：Flow cytometry is overexpressed influence of the taxol to apoptosis in nasopharyngeal carcinoma cells after ATF3.B：Clone shape
The influence that taxol is proliferated nasopharyngeal carcinoma cell after being overexpressed ATF3 is detected at experiment.C：After scratch experiment detection is overexpressed ATF3
Influence of the taxol to metastatic ability of nasopharyngeal carcinoma cells.Each group histogram data is the average value of independent repeated trials three times；
Bars indicates standard deviation, and * p ＜ 0.05, * * p ＜ 0.01, * * * p ＜ 0.001, TAXOL concentration for the treatment of is CNE-1/T IC25 Japanese yews
Determining alcohol.
Fig. 6：A：JASPAR software predictions ATF3 and MRVI1-AS1 promoter region binding sites have one close to sintering
And scoring is higher.B：GEO database analysis learns that the expression of ATF3 and MRVI1-AS1 are proportionate.C-E：QRT-PCR is examined
It surveys in CNE-1/T and is overexpressed or knocks out the expression of miR-513a-5p, miR-27b-3p, MRVI1-AS1 after ATF3.F：It is double
Luciferase detects MRVI1-AS1 promoter regions wild type and saltant type and ATF3 cotransfection 293T vehicles cells, detects fluorescence
Plain enzymatic activity.G：ChIP-PCR experimental verification MRVI1-AS1 promoter regions are combined with ATF3, and IgG is negative control group.Each group
Histogram data is the average value of independent repeated trials three times；Bars indicates standard deviation, * p ＜ 0.05, * * p ＜ 0.01.
Fig. 7：A-B：The expression of WB and qRT-PCR detections FRMD6, RASSF1, MOB1, TAZ in CNE-1 and CNE-1/T
Amount.C：ChIP-PCR experimental verification RASSF1 promoter regions are combined with ATF3, and IgG is negative control group.D：WB experiment detections
The expression of RASSF1 and TAZ under the conditions of different disposal.E：It is overexpressed MRVI1- in immunohistochemistry verification CNE-1/T transplantable tumors
The expression of RASSF1 and TAZ after AS1.F-G：MTT and Flow cytometry RASSF1 and TAZ are to taxol to nasopharyngeal carcinoma
The influence of proliferation and apoptosis.Each group histogram data is the average value of independent repeated trials three times；Bars indicates standard deviation, * p
＜ 0.05, * * p ＜ 0.01, * * * P ＜ 0.01.
Specific implementation mode
The present invention will be further explained with reference to the accompanying drawings and examples
Embodiment 1
MRVI1-AS1 can increase nasopharyngeal carcinoma chemosensitivity of pacilitaxel in vivo and in experiment in vitro
We carry out MTT detections to confirm the Japanese yew of tumour cell to CNE-1, CNE-1/T, HNE-2, HNE-2/T first
Alcohol drug resistance (Figure 1A), later to above four plants of cells row lncRNA cDNA microarrays, after screening, it has been found that MRVI1-AS1
Expression quantity is significantly lower than parent plant (Figure 1B) in two plants of persisters, and MTT results show to work as is overexpressed MRVI1- in persister
After AS1, CNE-1/T cells have obtained significant reverse to the drug resistance of taxol.Flow cytometry, Clone formation and cut are real
Test further demonstrate be overexpressed MRVI1-AS1 can increase nasopharyngeal carcinoma chemosensitivity of pacilitaxel (Fig. 1 C-F).In order to further
The function of MRVI1-AS1 is verified, we are by CNE-1, CNE-1/T and are overexpressed the CNE-1/T of MRVI1-AS1 to mouse progress
It is subcutaneously injected, according to grouping with or without taxol treatment, it has been found that compared with the control group, MRVI1-AS1 overexpression groups are used
Gross tumor volume smaller (Fig. 1 G-H) after taxol treatment.Tumor proliferation index of correlation Ki67, Ke Yifa are detected by immunohistochemistry
The Ki67 positive rates of existing MRVI1-AS1 overexpression groups are significantly less than control group (Fig. 1 I).No matter result above proves MRVI1-AS1
In vivo or it can increase nasopharyngeal carcinoma chemosensitivity of pacilitaxel in vitro.
Embodiment 2
It is quick that MRVI1-AS1 can adsorb miR-513a-5p and miR-27b-3p increase nasopharyngeal carcinoma Paclitaxel Chemotherapies by targeting
Perception
In order to further explore the mechanism that MRVI1-AS1 increases nasopharyngeal carcinoma chemosensitivity of pacilitaxel, we determine first
The positioning of MRVI1-AS1 in the cell finds it while being expressed in nucleus and cytoplasm (Fig. 2A).Then we to CNE-1,
CNE-1/T and CNE-1/T+MRVI1-AS1 is overexpressed three samples of strain and has carried out the detection of miRNA and mRNA chips, in miRNA
We have found that miR-27b-3p is overexpressed in strain in CNE-1/T+MRVI1-AS1 compared with expression quantity with miR-513a-5p in chip
CNE-1/T is decreased obviously (Fig. 2 B), and they simultaneously at the ends 3'UTR of MRVI1-AS1, there are possible binding site (Fig. 2 C).
In CNE-1 cells and CNE-1/T cells detect miR-27b-3p and miR-513a-5p expression, find they
Expression quantity in CNE-1/T cells is apparently higher than CNE-1 cells, and after being overexpressed MRVI1-AS1 in CNE-1/T cells,
The expression quantity of miR-27b-3p and miR-513a-5p is decreased obviously (Fig. 2 D) compared to control group, illustrates that MRVI1-AS1 can be born
To the expression of regulation and control miR-27b-3p and miR-513a-5p.We determine MRVI1-AS1 using Dual-Luciferase experiment
With the targeting relationship of miR-27b-3p and miR-513a-5p, miR-27b-3p analogies, miR-513a-5p analogies are divided
Not with MRVI1-AS1 wild types and saltant type cotransfection 293T vehicles cells, MRVI1-AS1 wild types miR-27b- is as a result shown
The Dual-Luciferase activity relative value and blank of 3p analogies group and MRVI1-AS1 wild type miR-513a-5p analogies groups
Group is compared with NC groups to be substantially reduced, and miR-27b-3p analogies and miR-513a-5p analogies are glimmering to MRVI1-AS1 saltant types
Light element enzymatic activity has no significant effect (Fig. 2 E).In addition, we additionally use RNApull down experiments further verification MRVI1-
As a result relationship between AS1 and miR-27b-3p and miR-513a-5p shows Bio-miR-513a-5p groups and Bio-miR-
The enrichment degree of MRVI1-AS1 is apparently higher than Bio-miR-NC, Bio-miR-513a-5p-Mut and Bio-miR- in 27b-3p groups
27b-3p-Mut groups (Fig. 2 G), the above result shows that MRVI1-AS1 can be targeted in conjunction with miR-27b-3p and miR-513a-5p simultaneously
Inhibit its expression.Then we use purple after inhibiting the expression of miR-27b-3p and miR-513a-5p in CNE-1/T cells
China fir alcohol handle cell, respectively row flow cytometry, colony formation, MTT and scratch experiment detect they to Apoptosis,
As a result the influence of proliferation and transfer is shown after inhibiting miR-27b-3p and miR-513a-5p respectively, the CNE- after taxol treatment
1/T apoptosis rates rise compared to control group detail, and proliferation rate and transfer ability are then then decreased obviously compared to control group
(Fig. 3 A-D).The above results show that MRVI1-AS1 can be thin to increase nasopharyngeal carcinoma by targeting miR-27b-3p and miR-513a-5p
Born of the same parents' paclitaxel-sensitive.
Embodiment 3
MRVI1-AS1 can be by restoring ATF3 expression quantity to increase nasopharyngeal carcinoma chemosensitivity of pacilitaxel
In order to further study the downstream target gene of MRVI1-AS1, miR-27b-3p and miR-513a-5p, we are right
The result of mRNA chips is analyzed, and combines GEO, TCGA database and miRNA target prediction tools (microRNA.org
And TargetScan), it is found that significantly returning occur in ATF3 expressions after CNE-1/T overexpressions MRVI1-AS1 in chip
It rises (Fig. 4 A)；And target spot prediction result shows that ATF3 exists simultaneously the binding site with miR-27b-3p and miR-513a-5p
(Fig. 4 B)；The expression that qRT-PCR results are shown in ATF3 after overexpression MRVI1-AS1 in CNE-1/T cells obviously rises
(Fig. 4 E), and after inhibiting the expression of miR-27b-3p or miR-513a-5p in CNE-1/T cells, the expression of ATF3 is same
Sample obviously rises (Fig. 4 F).This show miR-513a-5p and miR-27b-3p can negative regulation ATF3 expression, and MRVI1-
AS1 can promote the expression of ATF3, WB experiments to further demonstrate that MRVI1-AS1 can be by targeting miR-27b-3p and miR-
513a-5p raises the expression quantity (Fig. 4 C-D) of ATF3.In order to further confirm miR-27b-3p and miR-513a-5p and ATF3
Between relationship, we are tested using Dual-Luciferase, by miR-27b-3p analogies and miR-513a-5p analogies respectively with
ATF3 wild types and saltant type cotransfection 293T vehicles cells, as a result show ATF3 wild types miR-27b-3p analogies group and
The Dual-Luciferase activity of ATF3 wild type miR-513a-5p analogies groups is substantially reduced compared with blank group and NC groups, and
MiR-27b-3p analogies have no significant effect (Fig. 4 G) with miR-513a-5p analogies to ATF3 mutant luciferase activity,
This shows that miR-27b-3p and miR-27b-3p can be directly targeted AFT3.In order to further explore ATF3 and nasopharyngeal carcinoma taxol
Relationship between chemosensitivity, we are overexpressed AFT3 in CNE-1/T cells, with MTT after taxol treatment cell and gram
The grand proliferation rate for forming the CNE-1/T that experiment detection discovery is overexpressed ATF3 is significantly lower than control group (Fig. 4 H, Fig. 5 B)；Streaming is thin
The detection of born of the same parents' art finds that the apoptosis rate for being overexpressed the CNE-1/T cells after ATF3 is apparently higher than control group (Fig. 5 A)；Scratch removal is real
It tests detection and finds that the CNE-1/T transfer abilities for being overexpressed ATF3 are then significantly lower than control group (Fig. 5 C).The above results show
MRVI1-AS1 can be thin to increase nasopharyngeal carcinoma to restore the expression quantity of ATF3 by targeting miR-27b-3p and miR-513a-5p
Born of the same parents' chemosensitivity of pacilitaxel.
Therefore present invention discover that MRVI1-AS1 can pass through the expression water of targeted inhibition miR-513a-5p and miR-27b-3p
It puts down to raise the expression quantity of ATF3 to increase nasopharyngeal carcinoma chemosensitivity of pacilitaxel.
Embodiment 4
Transcription factor ATF3 can the positive expression for promoting MRVI1-AS1 in nasopharyngeal carcinoma
The target gene that ATF3 is miR-27b-3p and miR-513a-5p is we demonstrated in research before, and
MRVI1-AS1 can restore the expression of ATF3 by targeting miR-27b-3p and miR-513a-5p.But ATF3 as transcription because
Son can also be combined the expression to controlling gene with promoter.There is scholar to have been found that ATF3 has in kinds of tumors before
There is promotion Apoptosis, reduce the effect of transfer, therefore we guess whether ATF3 can be combined with the promoter of MRVI1-AS1
To regulate and control its expression.In order to verify this guess, we use gene database (http first://
Www.genome.ucsc.edu/ the base sequence of MRVI1-AS1 starting points upstream 1500) is found, then is combined by transcription factor
Site estimation database (http://jaspar.genereg.net/) it is predicted with transcription factor ATF3, it finds
The possible binding sites of ATF3 at 1432bp-1439bp, and score more a height of 7.145 points (Fig. 6 A).Pass through GEO databases point
Expression positive correlation each other in nasopharyngeal carcinoma of MRVI1-AS1 and ATF3 is learnt in analysis, this shows that ATF3 is possible to positive feedback
Adjust MRVI1-AS1 (Fig. 6 B).Next ATF3 is overexpressed in CNE-1/T cells, PCR detections find the table of MRVI1-AS1
Up to horizontal apparent increase, the expression of miR-27b-3p and miR-513a-5p declines, and the expression of ATF3 is inhibited then to obtain
Antipodal result (Fig. 6 C-E), this show the expression of ATF3 and MRVI1-AS1 be positively correlated and and miR-27b-3p
And the expression of miR-513a-5p is negatively correlated.It is double glimmering in order to further prove the relationship between ATF3 and MRVI1-AS1
Light element enzyme experimental result shows that ATF3 can be combined with MRVI1-AS1 promoter regions.In addition, we have also carried out ChIP-PCR
It is verified, is overexpressed ATF3 and silence ATF3 respectively in CNE-1/T cells, with ATF3 antibody sediment composite and takes out
DNA is carried, for MRVI1-AS1 promoter region design primers, the expression of row PCR detections MRVI1-AS1.As a result table was shown
Enrichment degree up to MRVI1-AS1 in the CNE-1/T cells of ATF3 obviously rises, and in the CNE-1/T cells of silence ATF3
The enrichment degree of MRVI1-AS1 is then decreased obviously (Fig. 6 G).The above result shows that transcription factor ATF3 can be opened with MRVI1-AS1
Mover combines and the expression of positive regulation MRVI1-AS1.
Embodiment 5
It is quick that MRVI1-AS1 may regulate and control HIPPO signal paths increase nasopharyngeal carcinoma Paclitaxel Chemotherapy by transcription factor ATF3
Perception
In order to further probe into the molecular mechanism that MRVI1-AS1 increases nasopharyngeal carcinoma chemotherapy sensibility, we are to mRNA chips
Further analysis is carried out.Ironically, in chip results there is not a series of HIPPO signal paths relevant molecule expression quantity
Same variation, middle and upper reaches regulatory factor RASSF1, FRMD6 and associated kinase MOB1 are overexpressed in CNE-1 and CNE-1/
Expression in MRVI1-AS1 groups is apparently higher than CNE-1/T groups, and the final effect molecule TAZ of HIPPO-TAZ signal paths
Expression trend it is then exactly the opposite.Chip results are verified by RT-PCR and WB, the wherein expression quantity of FRMD6 is in each group
Middle no significant difference, RASSF1 and the MOB1 expression quantity in CNE-1 groups are apparently higher than CNE-1/T groups, it should be noted that TAZ's
MRNA expressions no significant difference in each group, but CNE-1 groups are apparently higher than CNE-1/T groups (Fig. 7 A- on protein level
B), illustrating TAZ albumen, there are posttranslational modification effects in CNE-1 parent plants.Then we by Jaspar and ALGGEN into
Row analysis finds that ATF3 and RASSF1 promoter regions have a binding site close to translation initiation region, and two softwares have it is same
Site, FRMD6 promoter regions also have binding site with ATF3 but ATF3 overexpressions have no significant effect FRMD6 levels.MOB1
Promoter region is then without the higher ATF3 binding sites that score.ChIP experimental results further prove that ATF3 can positive regulation
The expression (Fig. 7 C) of RASSF1.Since RASSF1 is the upstream regulation factor of HIPPO, signal path, it is presumed that ATF3 whether
It can inhibit the expression quantity of TAZ by raising the expression of RASSF1, WB results, which are shown in CNE-1/T, is overexpressed ATF3
When TAZ expression quantity be significantly lower than control group, at the same time inhibit RASSF1 expression when TAZ expression quantity have significantly
Go up (Fig. 7 D), this shows that ATF3 can inhibit the expression of TAZ by promoting the expression quantity of RASSF1.And when we
The expression quantity of RASSF1 is significantly raised when being overexpressed MRVI1-AS1 in CNE-1/T and TAZ is then substantially reduced, while inhibiting ATF3
Expression after RASSF1 and TAZ expression trend it is then exactly the opposite (Fig. 7 D), immunohistochemistry also show be overexpressed MRVI1-AS1 when
The expression of RASSF1 rises and the expression of TAZ declines (Fig. 7 E).These are statistics indicate that MRVI1-AS1 can be by turning
Factors A TF3 is recorded to adjust HIPPO signal paths.Fluidic cell then shows that ATF3 can be inhibited by HIPPO accesses with MTT results
The expression of TAZ increases nasopharyngeal carcinoma chemosensitivity of pacilitaxel (Fig. 7 F-G).Above-mentioned the results show MRVI1-AS1 can lead to
It crosses ATF3 and regulates and controls HIPPO-TAZ signal paths to improve nasopharyngeal carcinoma chemosensitivity of pacilitaxel.
Claims (3)
1.ATF3 is preparing the application in treating nasopharyngeal carcinoma Paclitaxel Chemotherapy hypersitization medicine as target site.
2. application as described in claim 1, which is characterized in that the application refers to that ATF3 passes through tune as target site in preparation
It controls HIPPO accesses and increases nasopharyngeal carcinoma cell to the application in the drug of chemosensitivity of pacilitaxel.
3. application as claimed in claim 1 or 2, which is characterized in that the nasopharyngeal carcinoma refers to NPC.
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN110777204A (en) * | 2019-11-23 | 2020-02-11 | 中南大学湘雅三医院 | Application of MAFG-AS1 AS target site detection reagent in preparation of medicines for treating bladder cancer |
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2018
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Non-Patent Citations (3)
| Title |
|---|
| MARK M. WHITMORE ET AL: "Negative Regulation of TLR-Signaling Pathways by Activating Transcription Factor-3", 《THE JOURNAL OF IMMUNOLOGY》 * |
| ZHI-HUI YANG ET AL: "Cytokine and chemokine modification by Toll-like receptor polymorphisms is associated with nasopharyngeal carcinoma.", 《CANCER SCI》 * |
| 赵景娟等: "紫杉醇治疗晚期鼻咽癌的疗效分析与临床研究", 《中国医药指南》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110777204A (en) * | 2019-11-23 | 2020-02-11 | 中南大学湘雅三医院 | Application of MAFG-AS1 AS target site detection reagent in preparation of medicines for treating bladder cancer |
| CN110777204B (en) * | 2019-11-23 | 2023-04-07 | 中南大学湘雅三医院 | Application of MAFG-AS1 knock-out reagent in preparation of medicines for treating bladder cancer |
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