CN109528749A - Purposes of the long-chain non-coding RNA-H19 in preparation treatment hypophysis tumor medicine - Google Patents
Purposes of the long-chain non-coding RNA-H19 in preparation treatment hypophysis tumor medicine Download PDFInfo
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Abstract
The invention belongs to biomedicine technical fields, it is related to new application of the long-chain non-coding RNA-H19 in preparation treatment hypophysis tumor medicine, more particularly to long-chain non-coding RNA-H19 inhibits the effect of hypophysoma occurrence and development to go deep into Mechanism Study with it in vivo and in vitro, to achieve the purpose that treat hypophysoma.The present invention passes through cell culture, virus-mediated gene regulation experiment and the experiment of rat original position hypophysoma H19 injection model in vitro, the result shows that H19 reaches by inhibiting mTOR signal path downstream 4E-BP1 phosphorylation that growth of tumour cell is caused to block and inhibits growth of tumour cell purpose.Long-chain non-coding H19 of the invention can be used for preparing treatment hypophysis tumor medicine, is clinically used for treatment hypophysoma, can especially significantly inhibit tumour growth and achieve the effect that treatment.The present invention provides new strategy reference also for the especially intractable hypophysoma of clinical treatment hypophysoma.
Description
Technical field
The invention belongs to biomedicine technical field, the small molecule for being related to long-chain non-coding RNA (lncRNA)-H19 is medicinal
Purposes.More particularly to purposes of the H19 in preparation treatment hypophysis tumor medicine.
Background technique
It is investigated according to medical statistics, crowd's annual morbidity of China's pituitary adenoma is up to 7.5~15/,100,000 people, wherein just
Pituitary adenoma discovery rate is 10%~38.5% (average 22.5%) when the random mr imaging technique (MRI) of ordinary person group checks；
By 1,400,000,000 demographics of China human mortality quantity, annual new hair pituitary adenoma patients more than ten thousand people there are about 15-30, wherein prolactin pituitary
It is the most common hypotype in pituitary adenoma, about occupies 50% in all hypophysomas.Studies have shown that with medical technology progress,
Diagnostic level is increasingly advanced, the development of micrurgy and endoscopic technic, and hypophysoma incidence statistic is still in increase.Hypophysis
Adenoma causes the disorder of endocrine function, such as amenorrhoea-lactation and infertility, gigantism and acromegalia, and cushing syndrome etc.；With
And pituitary tumor increase compressing proximity structure leads to visual impairment and primary pituitary hypofunction etc., above-mentioned clinical symptoms seriously endanger
On the other hand the existence and quality of life of patient due to huge sufferer quantity, also brings great warp to country and patient home
The pressure for the pressure and medical resource of helping.
In clinical practice, prolactin pituitary choice drug treatment, wherein dopamine-receptor stimulant (Dopaminergic
Agonist, DA) it is that clinical first line therapy selects, China often uses bromocriptine, and American-European countries generally uses Cabergoline.It is above-mentioned
Drug enables to the patient of 80%-90% to effectively reduce gross tumor volume and restores normal prolactin(PRL (prolactin, PRL)
Level still about still has the patient of 10%-20% insensitive to bromocriptine even cabergoline treatment, in the industry by the portion
Point case is known as drug resistance case, and clinical practice is shown, even if the drug resistance patient, using operative treatment, the postoperative height that still faces is secreted
Newborn element mass formed by blood stasis can not restore the problems such as normal and tumor recurrence.In addition, other kinds of pituitary adenoma, such as growth hormone gland
The effective drug therapy of the shortages such as tumor, ACTH adenoma, TSH adenoma, quite a few patient belong to intractable hypophysoma；Therefore,
Drug resistance prolactinoma and intractable hypophysoma are still the huge difficult problem that the art clinic faces.
Long-chain non-coding RNA (long non-coding RNA, lncRNA) is that a kind of length is greater than 200 nucleic acid sequences
Single stranded RNA family, with controlling gene expressional function non-coding RNA.The study found that lncRNA takes part in Apoptosis tune
The processes such as control, tumor-infiltrated and transfer；In addition, the growth of tumour cell is also influenced by way of epigenetic regulation, therefore,
Important physiological action is played in tumor development and treatment.In recent years, research has found successively in tumor tissues
The lncRNA of some unconventionality expressions is related to the tumour of each system, and wherein lncRNA-H19 is earliest discovery and participates in tumour
One of important lncRNA of occurrence and development.H19 be insulin-like growth factor 2 (insulin-like growth factor2,
Igf2 the product of imprinted genes), is only expressed in embryonic tissue, is not expressed in adult tissue；It has been reported that and shows some swell
Tumor is related with the expression deletion of H19, such as Wilms tumor, bone marrow cell carcinoma, adrenal tumor.Meanwhile before present inventor
Phase is found and is verified in tissue specimen, H19 significant low expression in hypophysoma by biochip technology, illustrates that H19 may
Have the function of inhibiting hypophysoma growth.
Mammal rapamycin target protein (mammalian target of rapamycin, mTOR) is a kind of Soviet Union's ammonia
Acid/serine kinase, cell growth, differentiation and in terms of played important function.MTOR signal is not normal
Closely related, such as prostate cancer, lung cancer, gastric cancer and intracranial tumors such as glioma and hypophysoma occurs with tumour.MTOR suppression
Preparation be able to suppress due to the signal path extremely caused by tumour growth and vascularization, inhibit mTOR be used as clinical medicine
The foundation of object treatment malignant tumour.Therefore, mTOR research deeply has great importance to neoplasm targeted therapy.Activate mTOR
Multinomial cell function is participated in by certain factors in phosphorylated protein translation process, wherein mainly 4E-BP1 (eIF4E-
Binding protein1) and S6K1 (6 kinase 1 of ribosome protein subunit).4E-BP1 passes through competitiveness
Reach inhibition translation initiation in conjunction with eIF-4G and eIF-4E, and phosphorylation 4E-BP1 can speed up 4E-BP1 from eIF-4G with
It is dissociated on eIF-4E complex, to accelerate translation process.However, being related to grinding between lncRNA and mTOR signal path at present
Study carefully less, though having been reported that lncRNA-00961 can effectively inhibit mTOR access, and the regulation relationship between H19 and mTOR is still
It has no and has been reported that.
Status based on the prior art, present inventor is quasi- to provide the new pharmaceutical usage of long-chain non-coding RNA-H19,
More particularly to new application of the H19 in preparation treatment hypophysis tumor medicine.
Summary of the invention
The purpose of the present invention is the statuses based on the prior art, provide the new pharmaceutical usage of long-chain non-coding RNA-H19, tool
Body is related to new application of the H19 in preparation treatment hypophysis tumor medicine.
In the present invention, the long-chain non-coding RNA-H19 is 2369 length of nucleotides in rats, is in the mankind
2362 nucleotide, the two similarity 73%.
Long-chain non-coding RNA-H19 of the present invention can be grown with external effectively inhibition hypophysoma in vivo, through reality
Verifying is real, and the long-chain non-coding RNA-H19 is controlled by inhibiting the breeding of mTOR signal path blocks tumor cells to reach
Treat the purpose of hypophysoma.
In the embodiment of the present invention, in pituitary tumor cell system GH3 cell, h19 gene is overexpressed using slow virus and is carried out
In vitro test, the results show that it has an inhibiting effect to cell growth；Further Transplanted tumor model and tumor formation model card in situ
Real, H19 has the work for inhibiting its growth to the subcutaneous transplantation knurl model and rat prolactin secreting tumor cell growth in situ model of GH3 cell
With；Meanwhile H19 is better than Cabergoline to GH3 tumor formation inhibitory effect in vivo for subcutaneous transplantation knurl model experiment discovery；Mechanism is ground
Study carefully the result shows that: H19 is able to suppress hypophysoma and pituitary tumor cell growth, this kind of inhibiting effect is by inhibiting intracellular
The phosphorylation level of mTORC1 stream substrates 4E-BP1, to reach inhibition mTORC1 function to realize；
The present invention passes through cell culture, virus-mediated gene regulation experiment and rat original position hypophysoma H19 injection in vitro
Model experiment, the results showed that H19 causes growth of tumour cell to block by inhibition mTOR signal path downstream 4E-BP1 phosphorylation,
Inhibit growth of tumour cell purpose to reach；
It is of the invention the experimental results showed that, the long-chain non-coding H19 can be used for preparing treatment hypophysis tumor medicine, clinical
For treating hypophysoma, it can especially significantly inhibit tumour growth and achieve the effect that treatment.The present invention is also clinical treatment hypophysis
The especially intractable hypophysoma of tumor provides new strategy reference.
In order to make it easy to understand, the present invention will be described in detail by specific drawings and examples below.It needs
It is emphasized that specific example and attached drawing are merely to explanation, it is clear that those skilled in the art can be according to herein
Illustrate, various modifications and variations are made to the present invention within the scope of the invention, these modifications and variations are also included in this
In the range of invention.
Detailed description of the invention
Fig. 1 is shown: H19 is able to suppress hypophysoma GH3 cell, meanwhile, Nude Mouse Model confirms that H19 can be effective
Inhibit the subcutaneous tumor formation of GH3 cell.
Fig. 2 is shown: rat original position hypophysoma model is constructed using estrogen induction mode, it will using targeting injection technique
H19 is overexpressed adenoviral injection to rat pituitary tumor, and as a result H19 can effectively inhibit the growth of rat original position hypophysoma.
Fig. 3 is shown: by protein level experimental verification, H19 can effectively inhibit the phosphorylation level of 4E-BP1 to
Inhibit mTORC1 activity, and clear H19 by the phosphorylation of 4E-BP1 come regulating cell activity and GH3 cell tumor formation.
Fig. 4 is shown: being tested using nude mice by subcutaneous tumor formation, H19 compares Cabergoline, bright to the inhibiting effect of GH3 tumor formation
Aobvious enhancing, meanwhile, H19 is also better than Cabergoline to the phosphorylation level inhibiting effect of 4E-BP1.
Specific embodiment
Embodiment 1, H19 inhibit pituitary tumor cell strain inside and outside growth experiment
Experimental material:
(postposition culture solution is removed in operation for rat pituitary tumor cell strain GH3 (being purchased from ATCC) and people's primary pituitary adenoma cell
Directly send culture) in 37 DEG C, 5%CO2Under the conditions of routine culture, culture medium is containing 2.5% fetal calf serum (Gibco) plus 12.5%
Horse serum DMEM/F12 (Gibco).The slow virus for being overexpressed h19 gene is implemented in the lucky triumphant limited public affairs of biotechnology in Shanghai
Department,.MTS detects cell activity reagent and is purchased from Promega (U.S.).It is biological (China) that violet staining liquid is purchased from raw work.Nude mice
It purchases in SLAC (Shanghai).
Experimental method:
1) influence of the MTS experiment detection H19 for GH3 ability of cell proliferation
(1) the H19 GH3 cell being overexpressed and common control GH3 cell are suspended in conventional medium in advance, are adjusted
Cell density is 0.50 × 104Cells/well is inoculated in the adherent processing in 12 hours of 96 well culture plate, and every group of 5 secondary orifices, every hole is
10 microlitres of MTS detection reagent is added in 100 μ l systems；
(2) by cell in 37 DEG C, 5%CO2Under the conditions of cultivate 1-4 hours；With Detection wavelength 490nm measurement absorbance
Value, determines initial living cells quantity；
(3) cell surveyed is put back to 37 DEG C, 5%CO2Under the conditions of cultivate 24 hours, 48 hours, 72 hours, 92 hours；
(4) 10 microlitres of MTS detection reagents are added respectively at above-mentioned time point, detect numerical value by (2) method, calculates living cells
Quantity determines the relativity of two groups of cell activity, and N=5, experiment is parallel three times, and GH3 cell activity result is as shown in Fig. 1 .A.
2) influence that plate clone experiment detection H19 grows GH3 cell clone
(1) the H19 GH3 cell being overexpressed and common control GH3 cell are suspended in conventional medium, adjust cell
Density is that 500 cells/wells are inoculated in 6 well culture plates, and blank control group transfects the GH3 cell strain of blank slow virus；
(2) by above-mentioned cell culture in 37 DEG C, 5%CO2Under the conditions of, incubation time is 2-3 weeks；
(3) stop culture when naked eyes visible Clone formation every observation Clone formation situation on the 2nd during；
(4) violet staining liquid is used, is dyed 15 minutes at room temperature, clear water cleans dyeing liquor, takes pictures and leaves and takes as a result, picture is soft
Part counts clone's points, and experiment is parallel three times；Cell clonal formation situation such as Fig. 1 .B, shown in C.
3) Nude Mouse Model experiment determines that internal H19 influences GH3 cell tumor formation
(1) nude mice of 4 week old is chosen；
(2) the GH3 cell of the overexpression H19 of logarithmic growth and normal GH3 cell are collected, with serum free medium 10ml
In 1000 revs/min, it is centrifuged 3min, continuously washes 3 times, tumour cell finally is resuspended with serum free medium, density is adjusted to 1 × 107
Cell/ml, every nude mice oxter are inoculated with 100ul (i.e. 1 × 106A tumour cell), it can lay one's hand on and swell in nude mice oxter within 4 days or so
Tumor；
(3) daily with vernier calliper dipstick metering tumour long axis and wide axis size (unit: mm)；
(4) after raising 14-21 days, conventional treatment nude mice obtains knurl and weighs and take pictures, counts tumour weight and volume,
Gross tumor volume=long axis × wide axis2×1/2；In GH3 cell body, experimental result such as Fig. 1 .D, E, F shows；
Above-mentioned experimental result shows that H19 can inhibit outgrowth in hypophysoma GH3 cell body.
Embodiment 2, the experiment of rat original position hypophysoma H19 injection model
Experimental material:
The purchase of Fischer344 rat is tieed up experimental animal Co., Ltd, tonneau China in Beijing and is raised in Beijing Tiantan Hospital
Institute of neurosurgery；17- β estradiol is purchased from sigma company (U.S.)；The adenovirus construction of h19 gene is overexpressed in Shanghai
Han Heng biotech firm；
Experimental method:
H19 is to rat original position hypophysoma Experiment on therapy
(1) 4 week old female Fischer344 rats are chosen, 10, are divided into H19 treatment group and control group, every group 4；
(2) conventional every mouse is subcutaneously implanted the 17- β estrogen tablet containing 10mg, cultivates 6 weeks induced rat hypophysoma moulds
Type；Successfully building rat pituitary tumor model in situ is determined using magnetic resonance imaging (MRI) at the 6th week；
(3) it will be had under the guidance of micro- targeting injection technique with 10% chloraldurate (3.5ml/kg) anesthetized rat
H19 is overexpressed gene or the adenovirus of empty plasmid is injected in knurl respectively, and mode is using 10UI adenovirus (10^12 number
Magnitude)；
(4) continue culture 4-6 weeks, Magnetic Resonance Imaging measures tumor size, and anesthesia puts to death and takes sample；
Experimental result display H19 can be substantially reduced rat original position hypophysoma size, and (experimental result is united as shown in Fig. 2 .A
Figure is counted as shown in 2.B).
Embodiment 3, H19 are by inhibiting 4E-BP1 phosphorylation to influence cell activity
Experimental material:
RIPA lysate, PMSF, sample-loading buffer (5 ×), BCA protein quantification box, DAPI are purchased from green skies biotinylated biomolecule
Technical research institute (Jiangsu)；Protease Inhibitor Cocktail is purchased from Merck company (Germany).BSA is purchased from sigma
(9048-46-8)；PBS, 0.5%Trypsin-EDTA are purchased from Gibco；
Antibody: Tubulin (10068-1-AP, Proteintech), p-4EBP1 Thr70 (9455S, CST), 4E-BP1
(9644S, CST), p-4EBP1 Thr37/46 (9459S, CST), S6K1 (9202S, CST), p-S6K1 (9205S, CST), AKT
(9272, CST), p-AKT (9272, CST), strong type chemical luminescence reagent kit are purchased from Bio-rad company.
Experimental method:
1) cell protein is extracted detects with each destination protein
(1) GH3 cell is suspended in conventional medium, and adjusting cell density is 4 × 105Cells/well is inoculated in the training of 6 holes
Support plate in adherent overnight, every hole 2ml containing culture medium, final concentration of 25 μM of drug, respectively act on 0h, 12h, for 24 hours, 48h, 0h for pair
According to group, contain DMSO0.3%, in 37 DEG C, 5%CO2Under the conditions of cultivate；
(2) WesternBlot detects p-AKT, the expression of p-S6K1, p-4EBP1.With it is preceding by PMSF (100mM, 1: 100)
And ProteaselnhibitorCocktailSetIII (1: 200) is added RIPA lysate and collects carefully by drug treating time point
Born of the same parents remove culture solution in 6 orifice plates when collecting cell, after PBS gently washs cell, are digested GH3 cell 10 seconds with 0.05% pancreatin,
Pancreatin is removed, 1ml complete medium is added to neutralize, is collected in 1.5mlEP pipe, 2000rpm is centrifuged 5min, abandons supernatant, and 100 μ l are added and split
It solves liquid ice bath and cracks 30min；12000rpm is centrifuged 20min after cracking；It draws supernatant to be quantified, while taking part supernatant, add
Enter the sample-loading buffer (5 ×) of 1/4 volume, 100 DEG C are boiled 10min；Albumen after denaturation is stored in -80 DEG C of refrigerators；
(3) applied sample amount: the protein sample that BCA protein quantification box extracts each group quantifies, after correction on protein sample
Sample amount is 50 μ g；
(4) electrophoresis: the concentration of sample is carried out within 80V constant pressure electrophoresis 30 minutes；Carry out within 120V electrophoresis 1.5 hours point of sample
From judging electrophoresis degree according to marker；
(5) transferring film: wet turn, 170mA, constant current transferring film 2 hours is used；
(6) it antibody hybridization: is closed on room temperature shaker 1 hour with 5% BSA first；Confining liquid dilutes primary antibody
(Tubulin1:5000；p-AKT 1:2000；p-S6K11:1000；p-4EBP1 1:1000；), by the film to take a turn for the better in hybridization slot
In 4 DEG C of hybridized overnights；TBST washes film 3 times (room temperature shaker, each 2mL, 5min)；Secondary antibody is diluted with confining liquid, in hybridization slot
Middle room temperature shaker hybridizes 1 hour；TBST washes film and carries out chemiluminescence detection afterwards three times；
(7) detection of destination protein chemiluminescence detection: is carried out using enhanced chemical luminescence method (ECL method)；
Experimental result is as shown in Fig. 3 .A, 3.G.
2) histone extracts protein extraction and detection
(1) mouse is put to death after being anesthetized, strips off tumour with disscting instrument rapidly, and a little sample is taken to be put into 1.5ml cleaning EP
Guan Zhong is put into rapidly liquid nitrogen；
(2) after all samples take, the tissue specimen in liquid nitrogen is taken out, appropriate RIPA lysate is added；
(3) tissue homogenizer is fully ground tissue on ice, and 12000rpm is centrifuged 20min after cracking；Supernatant is drawn to be determined
Amount, while part supernatant is taken, the sample-loading buffer (5 ×) of 1/4 volume is added, 100 DEG C are boiled 10min；Albumen after denaturation saves
In -80 DEG C of refrigerators；
(5) it detects histone expression and points for attention is same as above, experimental result is as shown in Fig. 3 .B.
3) Nude Mouse Model experiment determines internal H19 by 4E-BP1 to inhibition hypophysoma occurrence and development
(1) nude mice of 4 week old is chosen；
(2) it by the normal GH3 cell of logarithmic growth, is overexpressed the GH3 cell of H19 and while being overexpressed and intervening 4E-
The GH3 of BP1 is collected, and with serum free medium 10ml in 1000 revs/min, is centrifuged 3min, is continuously washed 3 times, finally trained with serum-free
It supports base weight and hangs tumour cell, density is adjusted to 1 × 107Cell/ml, every nude mice oxter are inoculated with 100ul (i.e. 1 × 106A tumour is thin
Born of the same parents), it can be laid one's hand on and tumour in nude mice oxter within 4 days or so；
(3) daily with 3 groups of tumour long axis of vernier caliper measurement and wide axis size (unit: mm)；
(4) after raising 14-21 days, conventional treatment nude mice obtains knurl and weighs and take pictures, counts tumour weight and volume,
Gross tumor volume=long axis × wide axis2×1/2；Experimental result such as Fig. 3 .D, E, shown in F；
The experimental results showed that H19 is by 4E-BP1 to inhibition hypophysoma.
4) influence of the immunohistochemistry detection H19 to rat original position hypophysoma 4E-BP1 phosphorylation
(1) paraffin section de-waxing is to water；
(2) 3%H2O2Incubation at room temperature 5-10 minutes, to eliminate the activity of endogenous peroxydase；
(3) distilled water flushing, PBS impregnate 5 minutes x2 (if you need to antigen retrieval, carrying out after can walking herein)；
(4) 5%BS is closed, and is incubated at room temperature 10 minutes, incline serum deprivation, does not wash.Primary antibody working solution, 37 DEG C of incubation 1-2 are added dropwise
Hour or 4 DEG C overnight；
(5) PBS is rinsed, and 5 minutes x3 times.Be added dropwise appropriate biotin labeling secondary antibody working solution, 37 DEG C incubation 10-30 minutes,
PBS is rinsed, and 5 minutes x3 times；
(6) the strepto- avidin working solution of suitable horseradish enzyme or alkali phosphatase enzyme mark is added dropwise, 37 DEG C of incubation 10-30 divide
Clock, PBS are rinsed, and 5 minutes x3 times；
(7) DAB chromogenic reagent 10 minutes (DAB or NBT/BCIP)；
(8) tap water sufficiently rinses, and redyes, dehydration, and transparent, mounting shoots picture；Experimental result is as shown in Fig. 3 .C；
The experimental results showed that being able to suppress the phosphorylation of 4E-BP1 in H19 body.
Verifying H19 is better than Cabergoline to the inhibitory effect of hypophysoma in embodiment 4, nude mouse
Experimental material:
Nude mice purchasing channel is for example above-mentioned, Cabergoline desire to buy Tocris company (U.S.).
Test method:
1.H19 inhibits nude mice by subcutaneous GH3 tumor formation effect to be better than Cabergoline
(1) nude mice of 4 week old is chosen；
(2) by the normal GH3 cell of logarithmic growth, the GH3 cell for being overexpressed H19 is collected, with serum free medium 10ml
In 1000 revs/min, it is centrifuged 3min, continuously washes 3 times, tumour cell finally is resuspended with serum free medium, density is adjusted to 1 × 107
Cell/ml, every nude mice oxter are inoculated with 100ul (i.e. 1 × 106A tumour cell), it can lay one's hand on and swell in nude mice oxter within 4 days or so
Tumor；Wherein normal GH3 cell is divided to two groups, including control group and Cabergoline processing group；
(3) every other day stomach-filling Cabergoline is handled Cabergoline processing group, Drug level 0.75mg/kg；
(4) daily with 3 groups of tumour long axis of vernier caliper measurement and wide axis size (unit: mm)；
(5) after raising 14-21 days, conventional treatment nude mice obtains knurl and weighs and take pictures, counts tumour weight and volume,
Gross tumor volume=long axis × wide axis2×1/2；Experimental result such as Fig. 4 .A, B, shown in C.
2) H19 is better than Cabergoline to the inhibition of phosphorylation effect of 4E-BP1 in nude mouse
Histone extracting method detects normal group with above-mentioned respectively, H19 overexpression group, Cabergoline processing group, nude mice
The phosphorylation level of p-4E-BP1 in transplantable tumor, experimental result is as shown in Fig. 4 .D.
The present invention passes through cell culture, virus-mediated gene regulation experiment and rat original position hypophysoma H19 injection in vitro
Model experiment, the results showed that H19 causes growth of tumour cell to block by inhibition mTOR signal path downstream 4E-BP1 phosphorylation,
Inhibit growth of tumour cell purpose to reach.
Claims (6)
1. purposes of the long-chain non-coding RNA-H19 in preparation treatment hypophysis tumor medicine；
Described its length of nucleotides of long-chain non-coding H19 is 2369 in rats, is 2362 nucleotide, the two in the mankind
Similarity 73%.
2. purposes according to claim 1, which is characterized in that the long-chain non-coding RNA-H19 inhibits mTOR signal logical
The breeding of road blocks tumor cells, inhibits growth of tumour cell.
3. purposes as described in claim 2, which is characterized in that the tumour cell is subcutaneous transplantation knurl model GH3 cell
With rat prolactin secreting tumor cell original position induced growth cell.
4. purposes as described in claim 2, which is characterized in that the long-chain non-coding RNA-H19 inhibits intracellular mTOR
The phosphorylation level and then inhibition mTOR function of stream substrates 4E-BP1, realizes and inhibits hypophysoma and pituitary tumor cell growth.
5. purposes as described in claim 2, which is characterized in that the long-chain non-coding RNA-H19 inhibits GH3 cell Proliferation
Ability influences the growth of GH3 cell clone, inhibits outgrowth in hypophysoma GH3 cell body.
6. purposes according to claim 1, which is characterized in that the treatment hypophysis tumor medicine is for long-chain non-coding
The small-molecule drug of RNA-H19.
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| CN110302197A (en) * | 2019-07-26 | 2019-10-08 | 安徽医科大学第一附属医院 | A kind of lncRNA relevant to the miscellaneous Shandong amine therapeutic sensitivity of grace and its application in the miscellaneous Shandong amine treatment prostate cancer of grace |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5028588A (en) * | 1987-05-16 | 1991-07-02 | Somatogenetics International, Inc. | Blood substitutes |
| CN1404392A (en) * | 2000-02-22 | 2003-03-19 | 布里斯托尔-迈尔斯斯奎布公司 | Antiviral azaindole derivatives |
| US20070191490A1 (en) * | 2006-02-02 | 2007-08-16 | Sebti Said M | Withacnistin compounds for treatment of cancer |
| US20080102452A1 (en) * | 2006-10-31 | 2008-05-01 | Roberts Douglas N | Control nucleic acid constructs for use in analysis of methylation status |
| WO2010086388A1 (en) * | 2009-01-28 | 2010-08-05 | Ait Austrian Institute Of Technology Gmbh | Lung cancer methylation markers |
| CN103623429A (en) * | 2013-11-26 | 2014-03-12 | 广州中国科学院先进技术研究所 | Application of LincRNA (large intergenic non-coding RNA) in preparation of medicine for controlling tumor treatment drug resistance of methotrexate |
| CN106924735A (en) * | 2015-12-29 | 2017-07-07 | 上海交通大学医学院附属瑞金医院 | Purposes of the receptoroid activator of dopamine 1 in anti-tumor medicine is prepared |
-
2017
- 2017-09-22 CN CN201710877845.2A patent/CN109528749B/en active Active
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US5028588A (en) * | 1987-05-16 | 1991-07-02 | Somatogenetics International, Inc. | Blood substitutes |
| CN1404392A (en) * | 2000-02-22 | 2003-03-19 | 布里斯托尔-迈尔斯斯奎布公司 | Antiviral azaindole derivatives |
| US20070191490A1 (en) * | 2006-02-02 | 2007-08-16 | Sebti Said M | Withacnistin compounds for treatment of cancer |
| US20080102452A1 (en) * | 2006-10-31 | 2008-05-01 | Roberts Douglas N | Control nucleic acid constructs for use in analysis of methylation status |
| WO2010086388A1 (en) * | 2009-01-28 | 2010-08-05 | Ait Austrian Institute Of Technology Gmbh | Lung cancer methylation markers |
| CN103623429A (en) * | 2013-11-26 | 2014-03-12 | 广州中国科学院先进技术研究所 | Application of LincRNA (large intergenic non-coding RNA) in preparation of medicine for controlling tumor treatment drug resistance of methotrexate |
| CN106924735A (en) * | 2015-12-29 | 2017-07-07 | 上海交通大学医学院附属瑞金医院 | Purposes of the receptoroid activator of dopamine 1 in anti-tumor medicine is prepared |
Non-Patent Citations (6)
| Title |
|---|
| CONSTANCEVENNIN ET AL: "The long non-coding RNA 91H increases aggressive phenotype of breast cancer cells and up-regulates H19/IGF2 expression through epigenetic modifications", 《CANCER LETTERS》 * |
| ZE RUI WU ET AL: "Inhibition of mTORC1 by lncRNA H19 via disrupting 4E-BP1/Raptor interaction in pituitary tumours", 《NATURE COMMUNICATION》 * |
| 仲建平主编: "《中国外科年鉴》", 31 May 2012, 第二军医大学出版社 * |
| 刘凯: "长链非编码 RNA H19在侵袭性垂体腺瘤中的表达研究", 《中国优秀硕士学位论文全文数据库医药卫生科技辑》 * |
| 周德生主编: "《中医外治方全书 珍藏本》", 31 October 2015, 湖南科学技术出版社 * |
| 邱华等: "长链非编码ＲNAH19肿瘤性疾病关系的研究进展", 《山东医药》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110302197A (en) * | 2019-07-26 | 2019-10-08 | 安徽医科大学第一附属医院 | A kind of lncRNA relevant to the miscellaneous Shandong amine therapeutic sensitivity of grace and its application in the miscellaneous Shandong amine treatment prostate cancer of grace |
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