CN107432933A - Applications of the miR 22 as target site in the chemotherapy sensitizing medicine for preparing carcinoma of urinary bladder - Google Patents
Applications of the miR 22 as target site in the chemotherapy sensitizing medicine for preparing carcinoma of urinary bladder Download PDFInfo
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- CN107432933A CN107432933A CN201710568140.2A CN201710568140A CN107432933A CN 107432933 A CN107432933 A CN 107432933A CN 201710568140 A CN201710568140 A CN 201710568140A CN 107432933 A CN107432933 A CN 107432933A
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
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- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/337—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having four-membered rings, e.g. taxol
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
- G01N33/5008—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics
- G01N33/5011—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells for testing or evaluating the effect of chemical or biological compounds, e.g. drugs, cosmetics for testing antineoplastic activity
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
Abstract
Present invention demonstrates that miR 22 can target negativity regulation and control REDD1 expression, and the low expressions in Bladder Cancer of miR 22.MiR 22 can regulate and control axle by " REDD1 autophagy " influences sensitiveness of the transitional cell bladder carcinoma cell line to Paclitaxel Chemotherapy.Therefore the potential target spot that miR 22 may treat as Chemotherapy for Bladder Cancer enhanced sensitivity.Therefore, miR 22 can be used for the chemotherapy sensitizing medicine for preparing carcinoma of urinary bladder as target site.Especially miR 22 can be used for preparing as target site suppresses medicine of the autophagy increase transitional cell bladder carcinoma cell line to chemosensitivity of pacilitaxel.The carcinoma of urinary bladder refers to BUC.
Description
Technical field
The invention belongs to biomedicine technical field, and in particular to miR-22 is preparing the chemotherapy of carcinoma of urinary bladder as target site
Application in enhanced sensitivity medicine.
Background technology
BUC is most common malignant tumour in urinary system, has high relapse rate and high progression rates.Surgery excision is BUC
Standard treatments, but 50% Myometrial involvement BUC patient recur after surgery and in transferase 12 year it is dead.Combined chemotherapy energy
The recurrence rate of carcinoma of urinary bladder is reduced, improves survival.Microtubule stabilizer paclitaxel has been found to have well BUC
Curative effect.Recent three phases clinical trial finds to add on the basis of standard chemotherapy regimen cisplatin plus gemcitabine (GC)
Paclitaxel PCG schemes are added to effectively improve BUC general reaction rate and overall survival.But taxol resistance
Produce or patient is insensitive to taxol limits its clinical application.Therefore how to increase BUC is urgently to the sensitiveness of taxol
Clinical problem to be solved.
The activation of autophagy is one of important mechanisms caused by chemotherapy resistance, and it can assist tumour cell antagonism taxol etc. to change
The metabolic stress that medicine band is come is treated, so as to improve the survival rate of cell.REDD1 be stress GAP-associated protein GAP, anoxic, stress be with
A variety of DNA damages stimulate lower induced expression.REDD1 can participate in cell growth by suppressing mTOR signal paths, tumour is formed,
The regulation and control of cell ageing.Existing document report REDD1 plays oncogene in the tumours such as oophoroma, prostate cancer, non-small
Play tumor suppressor gene in the tumours such as cell lung cancer, breast cancer.But effects of the REDD1 in BUC there is no relevant report.REDD1
Induction autophagy can be also participated in by mTOR-EEF2K approach.Research report REDD1 induction autophagy promote myeloma cell,
The generation of prostate gland cancer cell chemotherapy resistance.But influence of the autophagy of REDD1 mediations to paclitaxel treatment BUC drug resistances
It is unclear.And regulation and control REDD1 mechanism is not known yet.
The content of the invention
Experiments indicate that REDD1 plays oncogene in BUC, its regulate and control the propagation of transitional cell bladder carcinoma cell line, apoptosis and
Autophagy.It is the potential treatment means for increasing transitional cell bladder carcinoma cell line chemosensitivity to suppress REDD1-EEF2K- autophagy axles.
Research report REDD1 expression and biological action have tissue specificity in different tumours.But REDD1 with
BUC patient clinicals pathology or the relation of prognosis there is no relevant report, and biological actions of the REDD1 in BUC is more unclear.This
Experiment finds versus normal tissues first, and REDD1 significantly increases in BUC tissue expressions.Further clinical analysis is shown
REDD1 high expression and the invasion and attack of BUC height, Lymph Node Metastasis and poor prognosis are closely related, and are BUC independent prognostic factors.Prompting
REDD1 is likely to become the potential index of BUC prognosis evaluations.
In order to further explore biological functions of the REDD1 in carcinoma of urinary bladder, we are in the high T24 cells for expressing REDD1
Middle suppression REDD1 expression, REDD1 is overexpressed in low expression REDD1 RT4 cells.It was found that wing after silence REDD1 expression
Guang cancer cell multiplication ability declines, apoptosis increase, is overexpressed proliferation of human bladder cancer cells ability increase after REDD1, and apoptosis is reduced.
Prompting REDD1 plays oncogene in transitional cell bladder carcinoma cell line.Our result is with REDD1 in oophoroma 15 , 16 And prostate cancer 17
In serve oncogene report it is consistent.REDD1 as stress GAP-associated protein GAP, can be by anoxic, nutritional deficiency, energy pressure etc.
Stressed condition excites.Tumour cell, especially advanced tumors cell are stored in above-mentioned all kinds of stress reactions, therefore tumour is thin
The expression that born of the same parents raise REDD1 is probably to escape a kind of self-protective mechanism of apoptosis.REDD1 is as the cancer base such as RAS, HIF-1 α
Regulation and control link, may suppress apoptosis and promote tumour progression by have activated anti-apoptotic program among cause.Research is found
The mechanism that REDD1 promotes cell survival may be relevant with activation AKT signal paths and suppression mTOR signal paths.
MTOR signal paths can adjust the activity of ULK1 protein activation compounds and then suppress autophagy.EEF2K is
Calcium/calmodulin-dependent enzyme, its up-regulated expression can trigger autophagy.Research finds mTOR signal paths
EEF2K degraded can also be promoted by phosphorylation EEF2K Ser78/Ser366 sites, and then it is horizontal to lower autophagy.And
REDD1 is MTOR inhibitor, can promote autophagy by the stable EEF2K of MTOR-EEF2K approach expression 20 .Therefore REDD1-
EEF2K- autophagy regulation and control axle is present.Cell EEF2K expression rises after REDD1 is overexpressed in this experiment, the horizontal up-regulation of autophagy, and
Knock out REDD1 and suppress EEF2K expression and autophagy level, further demonstrate above-mentioned regulation and control axle in transitional cell bladder carcinoma cell line first.
The autophagy of REDD1 mediations promotes the survival of cell.Therefore the autophagy that REDD1 promotes is probably that REDD1 plays cancer in carcinoma of urinary bladder
One of mechanism of gene action.
Appropriate activation autophagy can promote cell adapted stress and to promote to survive.Tumour cell autophagy increase is Chemoresistance
Or one of the reason for insensitive, therefore selective depression autophagy key regulatory molecule can be effectively increased chemosensitivity 10 , 33 .I
Find that in T24 cells silence REDD1 lowers autophagy after, sensitiveness increase of the cell to taxol；On in RT4 cells
After adjusting REDD1 activation autophagy, cell declines to the sensitiveness of taxol.Previous research also report that suppression REDD1- autophagy axles can
Promote myeloma cell, prostate gland cancer cell to the sensitiveness of chemotherapeutics.The taxol pair of IC25 dosage in this experiment
T24 and RT4 cell autophagies are horizontal and REDD1 expression has no significant effect (data are not presented).Taxol, which is reported, can promote ovum
The autophagy of nest cancer, lung carcinoma cell etc., but suppress the autophagy of MCF-7, SK-BR-3 breast cancer cell.Prompt taxol swollen in difference
Effect in oncocyte to autophagy is different, and the threshold value difference that autophagy may be triggered with the state and taxol of tumour cell is relevant.
In addition, present invention demonstrates that miR-22 can target negativity regulation and control REDD1 expression, and miR-22 is in Bladder Cancer
Middle low expression.MiR-22 can be by the way that " " regulation and control axle influences sensitiveness of the transitional cell bladder carcinoma cell line to Paclitaxel Chemotherapy to REDD1- autophagy.Cause
The potential target spot that this miR-22 may treat as Chemotherapy for Bladder Cancer enhanced sensitivity.
Therefore, miR-22 can be used for the chemotherapy sensitizing medicine for preparing carcinoma of urinary bladder as target site.Especially miR-
22 can be used for preparing medicine of the suppression autophagy increase transitional cell bladder carcinoma cell line to chemosensitivity of pacilitaxel as target site.It is described
Carcinoma of urinary bladder refers to BUC.
Brief description of the drawings
Fig. 1：Patient's poor prognosis of the high expression of REDD1.(A) immunohistochemical analysis REDD1 in 112 Bladder Cancers and
Expression in 32 normal bladder tissue paraffin specimens.(B) that 112 BUC patients are divided into REDD1 height expression group, REDD1 is low
Expression group, the life span difference that Kaplan-Meier curves methods analyze two groups of patients calculate P with log-rank test
REDD1 SABC representative graph in value (C) BUC tissues and normal structure.
Fig. 2：Propagation, apoptosis and the autophagy of REDD1 regulation and control BUC cells are horizontal.(A) WB detects bladder cancer cell lines
REDD1 expression in BIU87,5637, T24, EJ and RT4.(B) different shRNA-REDD1 (sh-1, sh-2, sh-3) transfections
T24 cells, REDD1 are overexpressed plasmid transfection RT4 cells.Ability of cell proliferation is detected with CCK-8 methods.(C) to intracellular LC3B
Fluorescence labeling is carried out, with the intracellular LC3B expressions of flow cytomery.(D) withering with flow cytomery cell
Die.(E) cell cycle of flow cytomery cell is used.(F) after intervening REDD1 expression, WB detections Caspase 3, c-
PARP, LC3-I/II, EEF2K expression.****P<0.0001, * * P<0.01, * P<0.05；
Fig. 3：REDD1 influences sensitiveness of the carcinoma of urinary bladder to taxol.(A) CCK-8 detects ability of cell proliferation.(B) WB is examined
Survey Caspase 3, c-PARP, LC3-I/II, REDD1 expression.
Fig. 4：MiR-22 Targeted-controls REDD1 expression.(A) microRNA.org software predictions miR-22 and REDD1 is deposited
In two potential binding sites.(B) QRT-PCR detects the expression of miR-22 in BIU87,5637, T24, EJ and RT4.(C)
QRT-PCR detections miR-22 expression after miR-22 mimics transfection T24 cells, anti-miR-22 transfection RT4 cells.(D)
After intervening miR-22 expression, WB detects REDD1 expression.(E) cotransfections of Lipofectamine 3000 are used in 293T cells
MiR-22 mimics and pMIR-REDD1-3 ' UTR-WT or pMIR- REDD1-3 ' UTR-MUT1or pMIR-REDD1-3 '
UTR-MUT2.Fluorescence activity is detected after transfection 48h. ****P<0.0001,***P<0.001, * * P<0.01；
Fig. 5：Propagation, apoptosis and the autophagy of miR-22 regulation and control transitional cell bladder carcinoma cell lines are horizontal.(A) miR-22 mimics transfect T24
And after miR-22 inhibitor transfections RT4, CCK-8 methods detection ability of cell proliferation；(B) flow cytomery cell is used
Apoptosis；(C) cell cycle of flow cytomery cell is used；(D) fluorescence labeling is carried out to intracellular LC3B, uses streaming
Cell instrument detects intracellular LC3B expressions；(E) after intervening miR-22 expression, WB detections Caspase 3, c-PARP, LC3-
I/II expression；
Fig. 6：MiR-22 influences sensitiveness of the carcinoma of urinary bladder to taxol.(A) CCK-8 detects ability of cell proliferation；(B) WB
Detect Caspase 3, c-PARP, LC3-I/II, REDD1 expression；
Fig. 7：Recover REDD1 expression can antagonism miR-22 mediation paclitaxel-sensitive.(A) WB detects Caspase 3, c-
PARP, LC3-I/II, REDD1, EEF2K expression；(B) CCK-8 detects ability of cell proliferation；(C) detection of drain cell instrument is thin
Born of the same parents' apoptosis；(D) flow cytomery cell LC3B is horizontal；
Fig. 8：Nude mice by subcutaneous is tested into knurl confirms that REDD1 and miR-22 influences sensitiveness of the tumour cell to taxol.
Embodiment
Sensitiveness of the carcinoma of urinary bladder to taxol can be increased by suppressing autophagy by lowering REDD1 expression
Collect 112 Bladder Cancers and the paraffin section row immunohistochemical analysis of 32 normal bladder tissues.As a result show
Show relative to normal bladder tissue, REDD1 significantly high expression (Figure 1A) in Bladder Cancer.Kaplan-Meier curves
Analysis finds high expression REDD1 bladder cancer patients poor prognosis (Figure 1B).
The high expression in BUC due to REDD1, and the high low expressions of REDD1 and patient's prognosis are closely related, we are further
Explore the influence for intervening REDD1 expression to bladder cancer cell lines propagation, apoptosis.We analyze bladder cancer cell lines first
REDD1 expression in BIU87,5637, T24, EJ and RT4, find in high invasiveness T24, EJ REDD1 expressions compared with
Height, expression is relatively low (Fig. 2A) in low invasiveness RT4, BIU87,5637.We select the T24 cells of the high expression of REDD1
REDD1 knockouts are carried out, selects the RT4 of REDD1 low expressions to carry out REDD1 overexpressions, as a result shows shRNA-REDD1-3 (sh-3)
REDD1 expression can be substantially knocked out, and plasmid REDD1 can significantly improve REDD1 levels (Fig. 2 B) in RT4.We use CCK-8
Detect ability of cell proliferation, flow cytomery Apoptosis.As a result it is shown in T24 after silence REDD1 expression, cell
Multiplication capacity declines, Apoptosis increase (P<0.001)；And after being overexpressed REDD1 in RT4 cells, ability of cell proliferation increases
Height, Apoptosis reduce (P=0.037) (Fig. 2 B, D).Apoptosis index Caspase 3 and cleaved PARP (c-PARP) exist
Increase is expressed after REDD1 silences, the expression downward (Fig. 2 F) after REDD1 overexpressions, REDD1 is further demonstrated and is suppressing wing
Effect in Guang cancer cell-apoptosis.The detection of cell cycle finds that REDD1 influences cell cycle progression (P<0.001, P=
0.002, respectively, Fig. 2 E).Increase we have discovered that LC3B expressions are relative relative to RT4 cells, in T24
It is high.The expression of LC3B expressions is lowered after silence REDD1 in T24, and LC3B expressions are expressed after REDD1 is overexpressed in RT4
Rise (Fig. 2 C).Prompting REDD1 also assists in the horizontal regulation and control of transitional cell bladder carcinoma cell line autophagy.WB detections find LC3-II expression water
Put down and decline after REDD1 expression is lowered, rise after REDD1 overexpressions, further demonstrate above-mentioned discovery (Fig. 2 F).Due to
REDD1 can induce autophagy by activating EEF2K20.Therefore we further have detected EEF2K expression.It was found that REDD1
Overexpression have activated EEF2K expression, and silence REDD1 can suppress EEF2K expression (Fig. 2 F), prompt thin in carcinoma of urinary bladder
" REDD1-EEF2K- autophagy " axle equally exists in born of the same parents.Above-mentioned experiment prompting REDD1 participates in carcinoma of urinary bladder propagation, apoptosis and autophagy
Regulation and control.
REDD1, which is reported, participates in the regulation and control of kinds cancer chemosensitivity.But whether REDD1 regulates and controls transitional cell bladder carcinoma cell line to purple
The sensitiveness of China fir alcohol is unclear.In order to probe into whether REDD1 works in taxol induced transitional cell bladder carcinoma cell line death is regulated and controled,
We transfect REDD1 silence expression plasmids in T24 cells, after REDD1 overexpression plasmids are transfected in RT4 cells, carry out
The processing of Taxol IC25 drug concentrations.CCK-8 method ability of cell proliferation, as a result shows：After the expression of REDD1 silences, Japanese yew is added
Alcohol is to the cytotoxicities of T24 cells, but REDD1 is overexpressed can suppress cytotoxicity of the taxol to RT4 cells to a certain degree
(Fig. 3 A).Further, in the T24 cells after taxol treatment after silence REDD1 expression, Caspase 3and c-PARP
Expression rise.Caspase 3 and the c-PARP up-regulation (Fig. 3 B) of REDD1 energy antagonism taxol induceds are overexpressed in RT4 cells.
Above-mentioned experimental result prompting REDD1 participates in the apoptosis in bladder of regulation and control taxol induced.
MiR-22 Targeted-controls REDD1 expression
In order to illustrate the upstream regulatory mechanism of REDD1 regulation and control transitional cell bladder carcinoma cell line biological functions, we use
The softwares such as microRNA.org, TargetScan may regulate and control the miRNAs of REDD1 expression to predict.It was found that REDD1-3 ' UTR
May target for modulation (Fig. 4 A) in the presence of 2 miR-22.Detected in bladder cancer cell lines BIU87,5637, T24, EJ and RT4
MiR-22 expression, finds miR-22 low expressions in high expression REDD1 T24, EJ cell, and low expression REDD1 RT4,
The relatively high expression (Fig. 4 B) of miR-22 in BIU87,5637 cells.Inverse relationship be present in prompting miR-22 and REDD1 expression.I
In T24 cells transfect miR-22 mimicis, in RT4 silence miR-22 express, as a result show miR-22 mimicis
The expression of miR-22 in T24 cells can substantially be increased, and anti-miR-22 can significantly inhibit the expression of miR-22 in RT4
(Fig. 4 C).MiR-22 mimicis can suppress T24 cells REDD1 expression simultaneously, and anti-miR-22 can promote RT4 cells
REDD1 expression (Fig. 4 D).Prompt miR-22 energy negativity regulation and control REDD1 expression.Further we utilize in 293T cells
Dual-Luciferase is tested to probe into whether miR-22 can be directly targeted regulation and control REDD1.MiR-22, which is overexpressed, can substantially suppress the
luciferase activitiesof the reporter genes with wild-type 3’-UTR(3’-UTR-WT)of
REDD1 and-UTR (3 '-UTR-MUT2) of of mutant-type 23 ' REDD1.But miR-22 is to mutant-type 1
3 '-UTR (3 '-UTR-MUT1) of REDD1 luciferase activity have no significant effect (Fig. 3 E).Prompt REDD1
The sites of 3 '-UTR mutant-type 1 are miR-22 specific regulatory control target spot.To sum up, miR-22 can Targeted-control REDD1
Expression.
Due to propagation, apoptosis and the autophagy of REDD1 regulation transitional cell bladder carcinoma cell lines, and miR-22 Targeted-controls REDD1 expression,
Whether so miR-22 also influences the above-mentioned biological behaviour of carcinoma of urinary bladderT24 cells are overexpressed cell propagation energy after miR-22
Power declines, apoptosis increase.And in RT4 cells after silence miR-22 expression, ability of cell proliferation lifting, apoptosis reduces (figure:
5A、B).The detection of cell cycle finds that miR-22 influences cell cycle progression (Fig. 5 C).Apoptosis index Caspase 3 and c-
PARP is overexpressed expression increase in miR-22, is expressed after the expression of miR-22 silences and lowers (Fig. 5 E).MiR-22 is prompted to participate in wing
The regulation and control of Guang cancer cell multiplication, apoptosis.The expression of LC3B expressions is lowered after miR-22 is overexpressed in further T24, and
The expression of LC3B expressions rises (Fig. 5 D) after silence miR-22 expression in RT4.WB detects LC3-II expression, as a result proves
MiR-22 expresses has negativity regulation relation (Fig. 5 E) with LC3-II levels.Above-mentioned experiment prompting miR-22 participates in carcinoma of urinary bladder and increased
Grow, the regulation and control of apoptosis and autophagy.
We explore miR-22 to influence of the taxol to transitional cell bladder carcinoma cell line cytotoxicity, as a result show：miR-22
Mimics adds toxicity of the taxol to T24, and silence miR-22 expression can suppress taxol to a certain degree to RT4's
Toxicity (figure:6A).Further, after taxol treatment T24 cells, transfection miR-22 mimics can promote Caspase
3and c-PARP expression.Regrettably silence miR-22 acts on unobvious to the Caspase 3 and c-PARP of taxol induced
(Fig. 6 B).Above-mentioned experimental result prompting miR-22 participates in the apoptosis in bladder of regulation and control taxol induced.
Because REDD1 and miR-22 can increase sensitiveness of the transitional cell bladder carcinoma cell line to Taxol, and miR-22 can Targeted-control
REDD1 expression, therefore We conducted functional rehabilitation experiment, explore miR-22 whether by suppress REDD1 expression and then
Promote sensitiveness of the transitional cell bladder carcinoma cell line to Taxol.Compared with having transfected miR-22 mimics T24, T24 cell cotransfections
After miR-22 mimics and REDD1 are overexpressed plasmid, REDD1 expression is gone up.And with having transfected miR-22 inhibitor's
RT4 cells are compared, and REDD1 is expressed in the cotransfection RT4 cells of miR-22 inhibitor and REDD1 silence expression plasmids
Decline.The change of EEF2K expression also changes over positive correlation (figure with REDD1 expression:7A).Transfected with miR-22 mimics
Group is compared, and miR-22 mimics and REDD1, which is overexpressed plasmid co-transfection group, can substantially suppress poison of the taxol to T24 cells
Property.Compared with miR-22 inhibitor transfection groups, miR-22 inhibitor and REDD1 silence expression plasmid cotransfection groups
Toxicity (Fig. 7 A-C) of the taxol to RT4 cells can substantially be increased.In addition, miR-22 mimics and REDD1 is overexpressed plasmid
Autophagy caused by cotransfection energy restored miR-22 mimics is horizontal to be declined, miR-22 inhibitor and REDD1 silences
Expression plasmid can suppress the autophagy (Fig. 7 A, D) of miR-22 inhibitor inductions.Further We conducted Clone formation
Experiment.It was found that taxol can substantially suppress the Clone formation number of T24 and RT4 cells, the miR-22 mimics in T24 cells
Suppression of the taxol to cell clonal formation can be increased, but REDD1 can reverse miR-22 mimics toxic action.In RT4
MiR-22 inhibitor can suppress the toxicity of taxol in cell, but silence REDD1 can reverse miR-22 inhibitor couple
The protective effect of cell.Summary experimental result, miR-22 is we demonstrated at least partially by Targeted-control REDD1's
Sensitiveness of the expression increase transitional cell bladder carcinoma cell line to Taxol.
Last animal model is used to assess influences of the REDD1 to chemosensitivity of pacilitaxel.With sh-REDD1 plasmids or
After miR-22 mimics lower REDD1 expression, T24 cells one-tenth knurl ability declines, and taxol is to T24 cell subcutaneous transplantation knurls
Lethal effect increase (Fig. 8 A, C).After plasmid or miR-22 inhibitor up-regulation REDD1 expression being overexpressed using REDD1,
The increase of RT4 cells one-tenth knurl ability, and taxol weakens (Fig. 8 B, D) to the lethal effect of RT4 cell subcutaneous transplantation knurls.
MiR-22 plays tumor suppressor gene in numerous tumours, can be by targetting the genes such as MMP14, SIRT1, Gal-9
Participate in the occurrence and development of the tumours such as regulation and control stomach cancer, clear-cell carcinoma, liver cancer.Our experiment is also demonstrated that miR-22 in carcinoma of urinary bladder
Play oncogene in cell.MiR-22, which is reported, participates in the regulation and control of osteosarcoma, colon cancer chemotherapy sensitiveness.We demonstrate that
MiR-22 can increase sensitiveness of the transitional cell bladder carcinoma cell line to taxol by suppressing REDD1- autophagy axle.This last experiment passes through body
Interior experiment further proves that the expression for suppressing REDD1 by RNAi or miR-22 can increase transitional cell bladder carcinoma cell line to the quick of taxol
Perception.Sensitiveness of the transitional cell bladder carcinoma cell line to taxol can be increased by suppressing REDD1 expression by miR-22.
Claims (4)
- Applications of the 1.miR-22 as target site in the chemotherapy sensitizing medicine for preparing carcinoma of urinary bladder.
- 2. application as claimed in claim 1, it is characterised in that the application refers to that miR-22 can be used for making as target site The standby autophagy that suppresses increases transitional cell bladder carcinoma cell line to the application in the medicine of chemosensitivity of pacilitaxel.
- 3. application as claimed in claim 1, it is characterised in that the carcinoma of urinary bladder refers to BUC.
- Applications of the 4.miR-22 in targeting negativity regulation and control REDD1 expression preparations are prepared.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111918968A (en) * | 2018-03-14 | 2020-11-10 | 贝斯以色列女执事医疗中心 | Inhibitors of Micro-RNA 22 |
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Non-Patent Citations (2)
| Title |
|---|
| JINGYU WANG ET AL: "Molecular mechanisms and clinical applications of miR-22 in regulating malignant progression in human cancer", 《INT J ONCOL》 * |
| NADINE HOUÉDÉ ET AL: "Targeting the genetic alterations of the PI3K–AKT–mTOR pathway: Its potential use in the treatment of bladder cancers", 《PHARMACOLOGY & THERAPEUTICS》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111918968A (en) * | 2018-03-14 | 2020-11-10 | 贝斯以色列女执事医疗中心 | Inhibitors of Micro-RNA 22 |
| CN111918968B (en) * | 2018-03-14 | 2023-11-24 | 贝斯以色列女执事医疗中心 | Inhibitors of Micro-RNA 22 |
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| Wang et al. | MicroRNA-874 inhibits cell proliferation and induces apoptosis in human breast cancer by targeting CDK9 | |
| Huang et al. | LncRNA GAS5-AS1 inhibits glioma proliferation, migration, and invasion via miR‐106b-5p/TUSC2 axis | |
| Meng et al. | miR-183-5p functions as a tumor suppressor in lung cancer through PIK3CA inhibition | |
| Ebrahimi et al. | MicroRNA-mediated redox regulation modulates therapy resistance in cancer cells: clinical perspectives | |
| Chen et al. | MiR-1297 regulates the growth, migration and invasion of colorectal cancer cells by targeting cyclo-oxygenase-2 | |
| Cai et al. | MicroRNA-146b-5p inhibits the growth of gallbladder carcinoma by targeting epidermal growth factor receptor | |
| Alfardus et al. | MicroRNA regulation of glycolytic metabolism in glioblastoma | |
| Wang et al. | MicroRNA-19a functions as an oncogenic microRNA in non-small cell lung cancer by targeting the suppressor of cytokine signaling 1 and mediating STAT3 activation | |
| Guo et al. | miR-503 suppresses tumor cell proliferation and metastasis by directly targeting RNF31 in prostate cancer | |
| Wang et al. | Negative regulation of PTEN by MicroRNA-221 and its association with drug resistance and cellular senescence in lung cancer cells | |
| Carlos-Reyes et al. | Biological adaptations of tumor cells to radiation therapy | |
| Rao et al. | MiR-219a-5p enhances cisplatin sensitivity of human non-small cell lung cancer by targeting FGF9 | |
| Adil et al. | Targeting Akt-associated microRNAs for cancer therapeutics |
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