CN103952406A - siRNA of targeting STAT3 gene for inhibiting human malignant brain glioma propagation and its expression vector and use - Google Patents
siRNA of targeting STAT3 gene for inhibiting human malignant brain glioma propagation and its expression vector and use Download PDFInfo
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Abstract
The invention discloses siRNA of a targeting STAT3 gene for inhibiting human malignant brain glioma propagation and its expression vector and use, and belongs to the technical field of bioengineering. STAT3-siRNA is introduced into malignant brain glioma cells, targetedly acts on mRNA of the STAT3 gene to block its translation process and effectively makes the STAT3 gene silent so that human malignant brain glioma targeting STAT3 gene expression is effectively inhibited, propagation activity of the malignant brain glioma cells is effectively inhibited and cell apoptosis is induced and thus malignant brain glioma cell growth is inhibited. The siRNA of targeting STAT3 gene for inhibiting human malignant brain glioma propagation can be used for preparation of human malignant brain glioma-resistant treatment drugs having high efficiency, fast rate and strong singularity.
Description
Technical field
The present invention relates to biotechnology, particularly a kind of siRNA and expression vector and application that suppresses the target STAT3 gene of people's malignant glioma propagation.
Background technology
STAT3 is the important member of IL-6/JAK-STATs Signal Transduction Pathways, about molecular weight 89KD, and when finding at first and being familiar with STAT3 and being the acute phase reaction from research cytokine IL-6 mediation, it is a kind of transcription factor, promotes many polygenic transcribing.Along with going deep into of studying, find that STAT3 not only participates in cytokine mediated acute phase reaction, and STAT3 was abnormal activation in the mankind's Several Kinds of Malignancy afterwards, the biological behaviour of modulate tumor cell, comprising pernicious cerebral glioma.
In recent years research discovery, some little double-stranded RNAs (double-stranded RNA, dsRNA) can impel specific gene mRNA degraded in body efficiently, specifically, lure that the phenotype of specific gene disappearance appears in cell into, and this technology is RNAi.In the initial period of RNAi effect, exogenous or endogenic double-stranded RNA is cut into the small molecules interference RNA fragment (small interfering RNAs, siRNAs) of 19～21bp by Dicer enzyme; SiRNA is combined rear formation RNA and is induced reticent mixture (RNA induced silencing complex, RISC) with ribozyme mixture subsequently; RISC is navigated on homologous mRNA transcript and is cut mRNA by base pairing after activating, thereby partially or completely suppresses the expression of goal gene.On the other hand, cell, under the effect of RNA RNA-dependent polysaccharase, can be primer, take mRNA as template by siRNA, synthesizes the complementary strand of mRNA, thereby makes mRNA also become dsRNA, then under the effect of Dicer enzyme, is again cracked into siRNA.These newly-generated siRNA also have the effect of bringing out RNAi, and by this chain reaction, intracellular siRNA increases greatly, has significantly increased the inhibition to genetic expression.
From the mechanism of action of RNAi, this gene expression regulation method belongs to the adjusting after transcribing and has clear superiority: RNAi gene inhibition definite effect, apply the effect that the content that micro-siRNA can make its coding Disease-causing gene product declines more than 90%, even can reach gene knockout; Secondly, RNAi suppresses to have strict sequence-specific, treatment with strong points, thereby application technique can suppress a plurality of target spots of same gene or a plurality of different genes and unlikely phase mutual interference simultaneously; Meanwhile, the effect of RNAi has the experimental study that compact cascade type scale effect and high-penetrability are more suitable for malignant tumour; In addition, even the specificity of RNAi recognition sequence also can produce accurately and effectively sealing effect to the oncogene being formed by wild-type point mutation on not impact of wild type gene.In to the experiment of kinds of tumor cells system, also confirm: in tumour cell, all exist the mechanism of RNAi, the RNAi that chemosynthesis, plasmid/virus or other modes mediate can suppress the expression of endogenous RNA effectively.
Summary of the invention
The present invention is exactly in order to solve the therapy of tumor problems such as glioma, take RNA interference mechanism as basis, by synthetic or expression plasmid sequence transcribe the little double-stranded RNA of STAT3 (siRNA) of formation can targeting in the mRNA of STAT3 gene, thereby block its translation process, reach the result that STAT3 genetic expression is reduced, and a kind of siRNA and expression vector and application of target STAT3 gene of the people's of inhibition malignant glioma propagation are provided.
The present invention realizes according to following technical scheme.
A siRNA who suppresses the target STAT3 gene of people's malignant glioma propagation, this siRNA has the effect that specificity suppresses malignant glioma cell proliferation, and its base sequence is: 5'-TGAAATCATCATGGGCTAT-3'.
The siRNA of the target STAT3 gene of described inhibition people malignant glioma propagation, is characterized in that: the gene order of this siRNA targeted human number is the STAT3 transcript of NM_003150, acts on the 2169th to 2188 of this gene order mRNA.
A kind of siRNA Lentiviral that suppresses the target STAT3 gene of people's malignant glioma propagation, it comprises siRNA as claimed in claim 1, have specificity and suppress people STAT3 genetic expression, specificity suppresses the effect of people's malignant glioma cell proliferation.
The siRNA Lentiviral of the target STAT3 gene of described inhibition people malignant glioma propagation, this expression vector includes pGC-LV carrier, and in its DNA sequence dna, virus expression carrier is slow virus.
The siRNA of the target STAT3 gene of described inhibition people malignant glioma propagation suppresses the application in people's malignant glioma medicine in preparation.
The siRNA Lentiviral of the target STAT3 gene of described inhibition people malignant glioma propagation suppresses the application in people's malignant glioma medicine in preparation.
Result shows: STAT3siRNA can successfully be transfected in glioma cell by slow virus body mediation, and the cellular metabolism after transfection STAT3siRNA is suppressed, and Growth of Cells slows down, and CCK8 absorption value obviously declines compared with control group and unloaded group; Western blot result shows: transfection total protein and the phosphorylated protein of cell STAT3 of STAT3siRNA express all suppressed; Flow cytometry experiment shows: STAT3-siRNA transfection after 96 hours apoptosis ratio compared with control group and unloaded group showed increased.
Like this, the present invention imports STAT3-siRNA after malignant glioma cell, targeting is in the mRNA of STAT3 gene, block its translation process, reticent STAT3 gene, effectively suppresses the genetic expression of people's malignant glioma target STAT3 effectively, can effectively suppress the proliferation activity of malignant glioma cell, cell death inducing, suppresses thereby reach the object that tumour occurs and grows.This siRNA and Lentiviral thereof can be used for preparing antitumor drug efficient, quick, high specificity.
Accompanying drawing explanation
Fig. 1 is U87 cell qRT-PCR method screening target spot histogram analysis figure;
Fig. 2 is U251 cell qRT-PCR method screening target spot histogram analysis figure;
Fig. 3 is that Western blot method validation LV-STAT3siRNA knocks out design sketch in U87 cell;
Fig. 4 is that Western blot method validation LV-STAT3siRNA knocks out design sketch in U251 cell;
Fig. 5 is that the enzyme of slow virus recombinant vectors is cut evaluation figure;
Fig. 6 is PGC-LV carrier collection of illustrative plates;
Fig. 7 is 96 hours egfp expression situations after U87 cell transfecting;
Fig. 8 is 96 hours egfp expression situations after U251 cell transfecting;
Fig. 9 is the expression of apoptotic proteins after Western blot method detection transfection U87 cell;
Figure 10 is the expression of apoptotic proteins after Western blot method detection transfection U251 cell;
Figure 11 is each time point CCK8 absorption value after U87 cell transfecting;
Figure 12 is each time point CCK8 absorption value after U251 cell transfecting;
Figure 13 is stream measuring apoptosis situation after the transfection of U87 control group;
Figure 14 is stream measuring apoptosis situation after the transfection of the unloaded group of U87;
Figure 15 is stream measuring apoptosis situation after the transfection of U87 experimental group;
Figure 16 is stream measuring apoptosis situation after the transfection of U251 control group;
Figure 17 is stream measuring apoptosis situation after the transfection of the unloaded group of U251;
Figure 18 is stream measuring apoptosis situation after the transfection of U251 experimental group;
Figure 19 is the base sequence of siRNA of the present invention.
Embodiment
Below in conjunction with drawings and Examples, the invention will be further described.
Embodiment mono-: disturb siRNA sequence design and the screening of people STAT3 genetic expression
1. siRNA sequence design
The present invention be take RNA perturbation technique as basis, utilizing NCBI Genbank retrieval gene order number is the full length sequence of the STAT3mRNA of NM_003150, with reference to siRNA principle of design, and use the secondary structure of RNA structure software simulation said target mrna, avoid self in conjunction with Jing district as far as possible, select the less sequence in pairing region, He Huan district, Ru Xian district.Filter out respectively the sequence that 5 siRNA are 19 Nucleotide, and bring them into BLAST genome database and compare, got rid of and other encoding sequence/EST homologies.And distinguished called after STAT3-siRNA-1, STAT3-siRNA-2, STAT3-siRNA-3, STAT3-siRNA-4, STAT3-siRNA-5(in Table 1) (synthetic by Shanghai Ji Kai company) as shown in figure 19.
Table 1 disturbs the siRNA sequence of people STAT3 genetic expression
| Numbering | Target sequence information | GC content (%) | Initiation site |
| STAT3-siRNA-1 | TGAAATCATCATGGGCTAT | 36.84% | 2169 |
| STAT3-siRNA-2 | GATCATAAGGTCAGGAGAT | 42.11% | 3170 |
| STAT3-siRNA-3 | ACAATCTACGAAGAATCAA | 31.58% | 458 |
| STAT3-siRNA-4 | TGACCAACAATCCCAAGAA | 42.11% | 1664 |
| STAT3-siRNA-5 | AAAGAATCACATGCCACTT | 36.84% | 361 |
2. filter out the best siRNA sequence of inhibition
2.1 materials and methods
2.1.1 transfection: utilize slow virus for carrier by siRNA transfection in U87 and U251 cell.Experiment is divided into six groups, is respectively siRNA-1, siRNA-2, siRNA-3, siRNA-4, siRNA-5, and control group, the MOI=5 of slow-virus transfection.
2.1.2 adopt qRT-PCR method to detect STAT3mRNA expression level: adopt TRIZOL reagent to extract cell total rna ,-80 ℃ of preservations, standby.After cell transfecting 96 hours, extract total RNA quantitative, the RNA reverse transcription that each experimental group is got isodose is cDNA, then pcr amplification STAT3 gene and GAPDH gene, STAT3 primer is: upstream sequence is 5'-CCAAGGAGGAGGCATTCG-3', and downstream sequence is 5'-ACATCGGCAGGTCAATGG-3'.GAPDH primer is: upstream sequence is 5'-TGACTTCAACAGCGACACCCA-3', and downstream sequence is 5'-CACCCTGTTGCTGTAGCCAAA-3'.PCR reaction system is: (in Table 2)
Table 2 pcr amplification STAT3 reaction system
The amplification condition of STAT3 is 95 ℃ of denaturations, 15s; 95 ℃ of each step sex change afterwards, 5s; Annealing is extended 60 ℃, 30s; Carry out altogether 45 circulations.In the extension stage, read light absorption value at every turn; After PCR finishes, 95 ℃ of sex change 1min, are then cooled to 55 ℃, make the abundant combination of DNA double chain.Since 55 ℃ to 95 ℃, each step increases by 0.5 ℃, keeps 4s, reads light absorption value simultaneously.
2.1.3 adopt Western blot to detect the protein expression of STAT3: by logarithmic phase U87 and U251 cell, when visual field cell grows to 80% fusion by 2 * 10
6individual/ware is inoculated in the Tissue Culture Dish of diameter 10cm, and observation of cell degree of mixing together carries out the transfection of slow virus while being 30～40%, by the cell extraction total protein to time point, according to the step of Western blot, detects STAT3, the expression level of p-STAT3.
3. result
3.1 qRT-PCR detect the inhibition that siRNA expresses STAT3mRNA: by histogram (seeing Fig. 1, Fig. 2), can be found out in U87 and U251 cell, and with respect to NC, KD1 couple
sTAT3striking of expressing subtracts that all to reach more than 65% be Effective target site, and the target sequence of KD1-KD5 is respectively: TGAAATCATCATGGGCTAT; TGAAATCATCATGGGCTAT; ACAATCTACGAAGAATCAA; TGACCAACAATCCCAAGAA; AAAGAATCACATGCCACTT, statistic data is in Table 3.
The analysis of statistical data of table 3 qRT-PCR method endogenous sieve target
3.2 Western blot detect the inhibition situation of siRNA to STAT3 protein expression: detect after transfection 96 hours STAT3 protein expression situation in U87 and U251 cell, in U87 and U251 cell, STAT3, p-STAT3(Tyr705, Ser727) expression of albumen obviously decline (seeing Fig. 3, Fig. 4).
Embodiment bis-: the structure of STAT3siRNA Lentiviral
1. disturb clone's preparation
(1) prepare competent cell
1. single bacterium colony of picking from the fresh flat board in 37 ℃ of incubators cultivation 16h, forwards in a 1L flask that contains 100ml LB or SOB substratum.In 37 ℃ of violent joltings, cultivate 3h(rotary shaker, 300 revs/min);
2. under aseptic condition, bacterium is transferred in aseptic, disposable, an ice-cold 50ml polypropylene tube, on ice, placed 10min, make culture be cooled to 0 ℃;
3. in 4 ℃, with 4000 revs/min of centrifugal 10min, reclaim cell;
4. pour out nutrient solution, will manage and be inverted 1min, the trace nutrient solution of final residual is flow to end;
5. the ice-cold 0.1mol/L CaCl of 10ml
2resuspended every part of precipitation, is positioned on ice bath;
6. in 4 ℃, with 4000 revs/min of centrifugal 10min, reclaim cell;
7. pour out nutrient solution, will manage and be inverted 1min, the trace nutrient solution of final residual is flow to end;
8. the ice-cold 0.1mol/L CaCl of 2ml for every 50ml initial incubation thing
2resuspended every part of cell precipitation;
9. cell is distributed into aliquot, be put in-70 ℃ frozen.
(2) annealing product connects into linearizing interference carrier (in Table 4)
Table 4 ligation system
| Reagent | Positive control (μ l) | From connecting contrast (μ l) | Connection group (μ l) |
| dd H 2O | 13 | 14 | 13 |
| 10XT4 Buffer | 2 | 2 | 2 |
| 50% PEG4000 | 2 | 2 | 2 |
| Enzyme is cut carrier DNA 100ng/ μ l | 1 | 1 | 1 |
| The double-stranded DNA 100 ng/ μ l of annealing | 1 | - | 1 |
| T4 DNA Ligase | 1 | 1 | 1 |
| Total | 20 | 20 | 20 |
Illustrate: positive control and certainly connect contrast, the carrier adding is consistent with connection group, but the annealing product that positive control adds is GAPDH gene (being with identical sticky end).
(3) transform
1. with cooling aseptic suction nozzle, from every kind of competent cell suspension, get 200 μ l and transfer in aseptic Eppendorf tube, every pipe adds 2 μ l connecting fluids, rotates gently to mix content, places 30min in ice;
2. pipe is put on the EP pipe support of putting well in the circulator bath of pre-heating to 42 ℃, exactly places 90s, do not shake EP pipe support;
3. fast pipe is transferred in ice bath, made the cooling l-2min of cell;
4. every pipe adds 800 μ l SOC substratum.With water-bath, substratum is heated to 37 ℃, then pipe is transferred on 37 ℃ of shaking tables, incubation 45min makes bacteria resuscitation;
5. the competent cell 150 μ l having been transformed is transferred on the LB nutrient agar of Amp resistance (100 μ g/mL);
6. flat board is placed in to room temperature until liquid is absorbed;
7. be inverted plate, in 37 ℃, cultivate 16h.
2. the PCR of positive colony identifies
By the constructed good virus 200 μ l of upper step, with viral DNA, extract test kit extraction viral DNA the viral DNA of extraction carried out to polymerase chain reaction,PCR (PCR): primer sequence is respectively:
STAT3 upstream: 5 '-GCCCCGGTTAATTTGCATAT-3 '
Downstream: 5 '-GTAATACGGTTATCCACGCG-3 '
Reaction system is as shown in table 5, and total amount is 20 μ l.PCR reaction conditions: 94 ℃ of denaturation 2min, 94 ℃ of sex change 30s, 60 ℃ of annealing 30s, 72 ℃ extend 30s, circulation 30 times, extend 72 ℃ of 6min eventually.1.0% agarose gel electrophoresis, with the shooting of gel imaging instrument, carries out image analysis, (as shown in Figure 5, Figure 6).
Table 5 slow virus DNA PCR reaction system
| Reagent | Volume (μ l) |
| ddH 2O | 12.4 |
| 5×Taq buffer | 4 |
| DNTPs （2.5mM） | 1.6 |
| Primer（+）（10μM） | 0.4 |
| Primer（-）（10μM） | 0.4 |
| Template | 1 |
| Taq polymerase | 0.2 |
Embodiment
three: STAT3-siRNA Lentiviral suppresses the experiment in vitro research of glioma
After the U87 of the STAT3-siRNA Lentiviral transfection glioma that embodiment bis-is built and U251 cell, test as follows:
3.1 Western blot detect the expression of apoptotic proteins: by logarithmic phase U87 and U251 cell, when visual field cell grows to 80% fusion by 2 * 10
6individual/ware is inoculated in the Tissue Culture Dish of diameter 10cm, observation of cell degree of mixing together carries out the transfection of slow virus while being 30～40%, by the cell extraction total protein to time point, according to the step of Western blot, detect the expression level (seeing Fig. 7,8, Fig. 9,10) of apoptotic proteins.
3.2 CCK8 methods are measured cell-proliferation activity: select U87 and the U251 cell of logarithmic phase, with 5 * 10
4the cell density of/ml is inoculated in 96 orifice plates, and experimental group, unloaded group and control group are after inoculation 24h, and transfection also continues to cultivate.After cultivating 7d, carry out CCK8 detection, microplate reader is measured the OD value (seeing Figure 11, Figure 12) under 450nm wavelength.
After 3.3 STAT3-siRNA transfections on apoptotic impact: by logarithmic phase U87 and U251 cell by 4 * 10
5the amount of/ml is inoculated in the 6 flat Tissue Culture Plates in hole, after 24h, carries out transfection, and every kind of processing of siRNA repeats 3 times.After transfection 96h, collecting cell, and process routinely cell.Add 5ul Annexin V-Alexa Fluor 647 and/or 5ul PI, mix, 4 ℃ of lucifuges are hatched 15 minutes, in 1 hour, with flow cytometer (BD company), measure.U87 and U251 cell add after LV-STAT3siRNA, control group, unloaded group, apoptosis (LR+UR quadrant in the figure) ratio of LV-STAT3siRNA are respectively U87(17.97 ± 0.24%, 18.26 ± 0.88%, 46.57 ± 1.63%), U251(15.94 ± 1.18%, 16.88 ± 0.17%, 39.34 ± 0.87%).Control group is compared unknown significance difference with unloaded group, compares with LV-STAT3siRNA group, and difference all has statistical significance (P < 0.01).Figure 13 is stream measuring apoptosis situation after the transfection of U87 control group, and Figure 14 is stream measuring apoptosis situation after the transfection of the unloaded group of U87, and Figure 15 is stream measuring apoptosis situation after the transfection of U87 experimental group; Figure 16 is stream measuring apoptosis situation after U251 contrast transfection, and Figure 17 is stream measuring apoptosis situation after the transfection of the unloaded group of U251, and Figure 18 is stream measuring apoptosis situation after the transfection of U251 experimental group.
Above-mentioned experimental result shows STAT3-siRNA-1st, and desirable STAT3 siRNA sequence is imported people's malignant glioma cell, can suppress the mRNA of this gene and the expression of albumen and also can suppress the proliferation activity of malignant glioma cell.Therefore, suppress siRNA and the Lentiviral thereof of people's malignant glioma cell STAT3 genetic expression and propagation, for suppress the application of people's malignant glioma medicine in preparation.
SEQUENCE LISTING
<110> Tianjin City No.1 Central Hospital
<120> suppresses siRNA and expression vector and the application of people's malignant glioma target STAT3 genetic expression and propagation
<130>DNA
<160>1
<170>PatentIn version3.5
<210>1
<211>19
<212>DNA
<213>2Ambystoma laterale x Ambystoma jeffersonianum
<220>
<221>1
<222>(1)..(19)
<400>1
tgaaatcatc atgggctat 19
Sequence table
SEQUENCE LISTING
<110> Tianjin City No.1 Central Hospital
<120> suppresses siRNA and expression vector and the application of the target STAT3 gene of people's malignant glioma propagation
<130> DNA
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 19
<212> DNA
<213> 2 Ambystoma laterale x Ambystoma jeffersonianum
<220>
<221> 1
<222> (1)..(19)
<400> 1
tgaaatcatc atgggctat 19
Claims (6)
1. a siRNA who suppresses the target STAT3 gene of people's malignant glioma propagation, is characterized in that: this siRNA has the effect that specificity suppresses malignant glioma cell proliferation, and its base sequence is: 5'-TGAAATCATCATGGGCTAT-3'.
2. the siRNA that suppresses as claimed in claim 1 the target STAT3 gene of people's malignant glioma propagation, it is characterized in that: the gene order of this siRNA targeted human number is the STAT3 transcript of NM_003150, act on the 2169th to 2188 of this gene order mRNA.
3. a siRNA Lentiviral that suppresses the target STAT3 gene of people's malignant glioma propagation, it is characterized in that: this expression vector comprises siRNA as claimed in claim 1, have specificity and suppress people STAT3 genetic expression, specificity suppresses the effect of people's malignant glioma cell.
4. the siRNA Lentiviral of the target STAT3 gene that described inhibition people malignant glioma is bred as claimed in claim 3, this expression vector includes pGC-LV carrier, and in its DNA sequence dna, virus expression carrier is slow virus.
5. the siRNA that suppresses the target STAT3 gene of people's malignant glioma propagation described in claim 1 suppresses the application in people's malignant glioma medicine in preparation.
6. the siRNA Lentiviral that suppresses the target STAT3 gene of people's malignant glioma propagation described in claim 3 suppresses the application in people's malignant glioma medicine in preparation.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108465108A (en) * | 2018-03-12 | 2018-08-31 | 上海宇玫博生物科技有限公司 | A kind of specific gene target spot prevented or treat glioma |
| CN113528523A (en) * | 2021-07-06 | 2021-10-22 | 天津医科大学总医院 | CRRNA (crribonucleic acid) of specific targeting F3-T3 fusion gene based on CRISPR-Cas13a system and application |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1733916A (en) * | 2005-07-01 | 2006-02-15 | 中国人民解放军第三军医大学 | siRNA for suppressing Stat3 gene expression and its preparation method |
| CN1778918A (en) * | 2005-07-01 | 2006-05-31 | 中国人民解放军第三军医大学 | SiRNA for inhibiting Stat3 gene expression and preparation thereof |
| CN102250953A (en) * | 2011-07-19 | 2011-11-23 | 马玲娣 | SiRNA lentivirus vector of human STAT3 gene and construction method thereof |
| CN102304538A (en) * | 2011-08-05 | 2012-01-04 | 汪运山 | Construction and screening as well as applications for siRNAs expression carrier of stomach cancer target STAT3 gene |
-
2014
- 2014-03-19 CN CN201410101962.6A patent/CN103952406B/en not_active Expired - Fee Related
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1733916A (en) * | 2005-07-01 | 2006-02-15 | 中国人民解放军第三军医大学 | siRNA for suppressing Stat3 gene expression and its preparation method |
| CN1778918A (en) * | 2005-07-01 | 2006-05-31 | 中国人民解放军第三军医大学 | SiRNA for inhibiting Stat3 gene expression and preparation thereof |
| CN102250953A (en) * | 2011-07-19 | 2011-11-23 | 马玲娣 | SiRNA lentivirus vector of human STAT3 gene and construction method thereof |
| CN102304538A (en) * | 2011-08-05 | 2012-01-04 | 汪运山 | Construction and screening as well as applications for siRNAs expression carrier of stomach cancer target STAT3 gene |
Non-Patent Citations (3)
| Title |
|---|
| 李光辉等: "RNAi 抑制STAT3 表达、活化诱导人胶质瘤干细胞凋亡与分化", 《中华神经外科疾病研究杂志》 * |
| 王永志等: "STAT3 在恶性脑胶质瘤中研究进展", 《军医进修学院学报》 * |
| 袁小鹏等: "STAT3 抑制剂WP1066 对乏氧人脑胶质瘤细胞U251放射敏感性的影响", 《辐射研究与辐射工艺学报》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108465108A (en) * | 2018-03-12 | 2018-08-31 | 上海宇玫博生物科技有限公司 | A kind of specific gene target spot prevented or treat glioma |
| CN108465108B (en) * | 2018-03-12 | 2020-07-07 | 上海宇玫博生物科技有限公司 | Specific gene target for preventing or treating brain glioma |
| CN113528523A (en) * | 2021-07-06 | 2021-10-22 | 天津医科大学总医院 | CRRNA (crribonucleic acid) of specific targeting F3-T3 fusion gene based on CRISPR-Cas13a system and application |
| CN113528523B (en) * | 2021-07-06 | 2023-03-07 | 天津医科大学总医院 | CRRNA (crribonucleic acid) of specific targeting F3-T3 fusion gene based on CRISPR (clustered regularly interspaced short palindromic repeats) -Cas13a system and application of CRRNA |
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