CN104694477B - A kind of recombinant HEK 293 cell of EphrinB2 high expression and its application - Google Patents
A kind of recombinant HEK 293 cell of EphrinB2 high expression and its application Download PDFInfo
- Publication number
- CN104694477B CN104694477B CN201510094921.3A CN201510094921A CN104694477B CN 104694477 B CN104694477 B CN 104694477B CN 201510094921 A CN201510094921 A CN 201510094921A CN 104694477 B CN104694477 B CN 104694477B
- Authority
- CN
- China
- Prior art keywords
- ephrinb2
- cell
- recombinant
- expression
- recombinant hek
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
Links
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
Recombinant HEK 293 cell and its application, the recombinant cell the invention discloses a kind of EphrinB2 high expression are using HEK293 cells as host cell, and exogenous expression's carrier of transfection host cell is the expression vector for including EphrinB2 full-length genes.Compared with the existing recombinant cell built containing only EphrinB2 extracellular regions or partial gene sequence, expression EphrinB2 molecules that the recombinant HEK 293 cell film comprising EphrinB2 full length genes that the present invention is built can be stablized, and there is complete molecular activity.Mtt assay detection Dasatinib under a certain concentration plays the role of recombinant HEK 293 cell good, therefore can be applied in using EphrinB2 as the drug screening of target spot.
Description
Technical field
The invention belongs to biological technical field, it is related to the structure of the non-cancerous cells of EphrinB2 high expression, more particularly to one
The recombinant HEK 293 cell of EphrinB2 high expression of the kind based on external source recombinant plasmid and its application.
Background technology
Tyrosine kinase receptor (RTK) is a huge family, is taken in the vital movement of organism very heavy
The effect wanted.The extracellular region of RTK is binding partner domain, and ligand is soluble or film combination polypeptide or protein hormone,
Including insulin and a variety of growth factors.Intracellular section is the catalytic site of tyrosine protein kinase, and has autophosphorylation site.
After ligand is combined with RTK, receptor conformation change is can induce, causes acceptor that stable dimerization, the acceptor hair of dimerization occurs
Raw phosphorylation, forms the motif containing phosphorylated tyrosine site in receptor kinase area, and with the activation of PTK.Tyrosine
Special phosphorylation makes kinases area stabilization in the conformation of activation, and contains src Oncogene family SH2 areas and with reference to phosphoric acid for downstream
The signal transducers for changing the functional areas (phosphotyrosine binding domain, PTB) of tyrosine provide identification, stop
The position leaned on and combined.PTK adjusts growth, differentiation, movement, deformation and the metabolism of cell by the signal path in active cell
Deng causing a variety of effects.
Eph be belong to be currently known maximum protein tyrosine kinase receptor family (RTKs) subtribe, including Eph acceptors and
Its Ephrin ligand.EphB4 and EphrinB2 shows the mode of uniqueness when signal transmits, and can mutually be activated by other side, produces
" forward signal " and " reverse signal ", using signal paths such as EphrinB2 activation EphB4 as forward signal；And EphB4 is activated
EphrinB2 is reverse signal.EphB4/EphrinB2 and its transduction of the two-way signaling of uniqueness almost participate in each of angiogenic growth
Aspect.EphrinB2 may play the part of the role of a total regulation and control person in VEGF signal paths, and core is played in angiogenic growth
The heart acts on.Eph/Ephrin families are the maximum subtribes in receptor tyrosine kinase family, are divided into two according to ligand structure difference
Kind hypotype：The referred to as EphrinA hypotypes being fixed on after birth, totally 5 kinds；And the Ephrin ligands of 3 kinds of subtype Bs then belong to cross-film egg
(Cell, 1997,90 (3) in vain:403-404).Eph acceptors are also classified into two groups according to homologous sequence:EphA and EphB acceptors.
Eph acceptors are made of membrane-spanning domain, extracellular ligand identification domain and domain cysteine, are located at cell containing one in domain cysteine
The PDZ binding domain of inner side；Eph transmembrane proteins can be with positioned at the extracellular domain for being responsible for signal in identification cellular environment
Ephrin has an effect, and forms the form of polymer, thus causes the activation of Ephrin ligands and Eph acceptors, so as to influence thin
Born of the same parents interact and the function such as cell migration (Cur Opini Cell Bio, 2010,22:611-616).
Generation, development and the transfer of tumour depend on Tumor angiogenesis, and only after new vessels is formed, tumour is
It can mushroom out, and distally be shifted further to surrounding wetting and into blood circulation, and Eph/Ephrin signal transductions are
The important cells morphology Control factors of embryo growth, angiogenic growth and generating process, several in Eph/Ephrin families match somebody with somebody
Body and acceptor have confirmed take part in Tumor Growth (Cancer Cell, 2010,17:533-534).
Innovation drug research and the great research field that exploitation is in Country science and technology plan, antitumor drug targeting screening are
One important directions of new medicament screen and field.At present, finding target developing anti-tumor medicaments, oneself becomes treatment malignant tumour life
One of long and transfer important means.In the research and development of inhibiting tumour cells agent, there is not certain suppression to EphrinB2 still
Drugs with function.Using EphrinB2 as target spot, the medicine that searching targets EphrinB2 in tumour can become research and development antitumor drug
One of important channel.Research at present to EphrinB2 genes is more, but structure to full length sequence EphrinB2 carriers and
Biological applications research and its application have no report.
The content of the invention
Recombinant HEK 293 cell and its application it is an object of the invention to provide a kind of EphrinB2 high expression, are building
On the basis of the expression vector of EphrinB2 full-length genes, transfected HEK 293, is stablized, high expression EphrinB2 molecules
Recombinant HEK 293 cell strain, can be used in the screening using EphrinB2 as the medicine of target spot.
To reach above-mentioned purpose, the technical solution adopted by the present invention is：
A kind of recombinant HEK 293 cell of EphrinB2 high expression, which is thin as host using HEK293 cells
Born of the same parents, exogenous expression's carrier of transfection host cell are the expression vectors for including EphrinB2 full-length genes.
The nucleotide sequence of the EphrinB2 full-length genes is as shown in SEQ.ID.NO.1.
The expression vector comprising EphrinB2 full-length genes is pEZ-M61-EphrinB2, wherein pEZ-M61-
EphrinB2 is that EphrinB2 full-length genes are cloned into carrier pEZ-M61.
The primer sequence of the pEZ-M61-EphrinB2 is：
Sense primer：5-TCATCTTCATCGTCATCATCATC-3
Anti-sense primer：5-CTGTCCGCAGTCCTTAGC-3.
The recombinant HEK 293 cell of EphrinB2 high expression is using EphrinB2 as the application in the drug screening of target spot.
The medicine using EphrinB2 as target spot is the medicine that can be combined with acceptor EphB4-Fc.
Relative to the prior art, beneficial effects of the present invention are：
1st, the present invention provides a kind of recombinant cell of EphrinB2 high expression, in the table of structure EphrinB2 full-length genes
Up on the basis of carrier, transfected HEK 293, is stablized, the recombinant HEK 293 cell of high expression EphrinB2 molecules.With
The existing recombinant cell built containing only EphrinB2 extracellular regions or partial gene sequence is compared, and what the present invention was built includes
The expression EphrinB2 molecules that the recombinant HEK 293 cell film of EphrinB2 full length genes can be stablized, highest expression EphrinB2
29.3 times are improved before relatively being transfected by the scale of construction, and there is complete molecular activity.
2nd, the HEK293 cells as host cell are the immortality that primary human embryonic kidney cell transfects 5 type adenovirus (Ad5) DNA
Change cell, the various expression of receptor amounts which contains are relatively low；And recombinant HEK 293 cell provided by the invention
The relative expression quantity of EphrinB2 ligands is more obvious than HEK293 to be increased, relative to other cell surface molecules with occupying advantage of expression
Position, is on a good wicket in terms of its acceptor is combined；Thus substantially increase spirit of the screening using EphrinB2 as target drug
Quick property and specificity；Can be that further the validity of research EphrinB2 biological characteristicses and discovery feature albumen carries at the same time
For preferable cell model.
3rd, in the research and development of inhibiting tumour cells agent, the medicine of EphrinB2 is not targeted still.Therefore with structure
Recombinant HEK 293 cell has carried out Carbazole alkaloid experiment to the Sorafenib and Dasatinib medicine of listing, finds in a certain concentration
Lower Dasatinib has recombinant HEK 293 cell good inhibitory action, shows that recombinant HEK 293 cell provided by the invention can be with
Using the drug screening using EphrinB2 as target spot.
Brief description of the drawings
Fig. 1 is the plasmid map of pEZ-M61-EphrinB2 carriers；
Fig. 2 is the culture figure that DMEM culture mediums add G418 screening positive cells；Wherein a is control group HEK293 cells, and b is
Recombinant HEK 293 cell, c are the fluorograms of recombinant HEK 293 cell；
Fig. 3 is the RT-PCR detection figures of the EphrinB2 expressions of recombinant HEK 293 cell；Wherein 1 is restructuring HEK293
Cell, 2 is transfect the HEK293 cells of empty plasmid, and 3 be the control group HEK293 cells of untransfected；
Fig. 4 is agarose gel electrophoresis figure after the extracted RNA of recombinant HEK 293 cell, reverse transcription and PCR；Wherein swimming lane 1
For Marker, swimming lane 2 is the recombinant HEK 293 cell of transfection pEZ-M61-EphrinB2, and swimming lane 3 is that the HEK293 of untransfected is thin
Born of the same parents, swimming lane 4 are the HEK293 cells of transfection empty carrier；
Fig. 5 is that observation green fluorescent protein Qualitative Identification EphrinB2 is thin in restructuring HEK293 under inverted fluorescence microscope
Expression figure in born of the same parents；Wherein a is control group HEK293 cells, and b is recombinant HEK 293 cell；
Fig. 6 is the combination figure of recombinant HEK 293 cell and the acceptor EphB4 of EphrinB2；
Fig. 7 is the EphrinB2 expression spirograms of Western Blot detection recombinant HEK 293 cells；Wherein 1 is restructuring
HEK293 cells, 2 is transfect the HEK293 cells of empty plasmid, and 3 be the HEK293 cells of untransfected；
Fig. 8 is MTT experiment detection Dasatinib to the inhibitory action of recombinant HEK 293 cell and the amount effect curve figure of drafting.
Embodiment
The present invention is described in further details below in conjunction with the accompanying drawings.
The present invention constructs the carrier for expression of eukaryon pEZ-M61-EphrinB2 of EphrinB2 full-length genes, its gene first
Sequencing result is consistent with gene pool people EphrinB2, pEZ-M61-EphrinB2 carrier lipid body method transfected HEK 293s, warp
Cross screening is stablized, the recombinant HEK 293 cell of high expression EphrinB2 molecules, i.e. restructuring HEK293-EphrinB2 is thin
Born of the same parents.Elaborate below to present aspect is explanation of the invention rather than restriction.
A. the present invention is that primer sequence is as follows using pEZ-M61-EphrinB2 as basic carrier (GeneCopoeia companies)：
Sense primer：5-TCATCTTCATCGTCATCATCATC-3
Anti-sense primer：5-CTGTCCGCAGTCCTTAGC-3
The plasmid map of the pEZ-M61-EphrinB2 carriers of structure is as shown in Figure 1.
B.pEZ-M61-EphrinB2 expression vector transfected HEK 293s
Plasmid is transferred to HEK293 cells using liposome 2000, and (liposome 2000 is purchased from U.S. GIBCO, Invitrogen
Company, operates according to 2000 kit specification of liposome), DMEM culture mediums (Dulbecco's Modified Eagle
Medium after) adding 500 μ g/mL of G418 concentration to screen 14 days, obtained positive colony is under inverted fluorescence microscope with green
Color fluorescence, picking G418 resistant clones secondary cultures, adjust the DMEM culture mediums that G418 concentration is 300 μ g/mL and train after 4 weeks
Support, obtain the clonal cell line of the Positive transfections containing pEZ-M61-EphrinB2.
DMEM culture mediums add G418 to screen positive cell after 14 days, HEK293 cell culture figure such as Fig. 2 a institutes as control
Show；The cell culture figure of recombinant HEK 293 cell, i.e. HEK293-EphrinB2 positive cells is as shown in Fig. 2 b, 2c；Untransfected matter
The HEK293 complete cell deaths of grain pEZ-M61-EphrinB2, transfection have the HEK293 cellular portions of pEZ-M61-EphrinB2
Death, while survivaling cell carries green fluorescence under inverted fluorescence microscope, this contains anti-G418 resistances with transfected plasmids
Gene is related with green fluorescent protein GFP genes, shows that plasmid pEZ-M61-EphrinB2 is successfully transferred to host cell HEK293.
C. the RT-PCR of transfection cell strain is detected：
Untransfected and the cell total rna of transfection are extracted using TRIZOL kits respectively, and used by template of total serum IgE
The cDNA synthetic agent box of TAKARA companies carries out reverse transcription, obtains cDNA；
Respectively using gained cDNA as template, PCR amplification EphrinB2 full length genes.
PCR reaction systems are： Premix Ex TaqTMII 12.5μL；Sense primer (sense, 20 μm of ol/
L)0.5μL；Anti-sense primer (antisense, 20 μm of ol/L) 0.5 μ L；2 μ L of template (cDNA)；ddH2O9.5μL；Total：25μ
L。
PCR reaction conditions：95 DEG C of pre-degeneration 30s, 95 DEG C of denaturation 5s, 60 DEG C of extension 30s, are circulated 40 times；RT-PCR is detected
The results are shown in Figure 3, and 1 is recombinant HEK 293 cell, and 2 is transfect the HEK293 cells of empty plasmid, and 3 is thin for the HEK293 of untransfected
Born of the same parents make negative control；Compared with 2,3, EphrinB2 gene levels are significantly raised in 1, and highest is expressed EphrinB2 and relatively turned by the scale of construction
29.3 times are improved before dye.GAPDH groups are the positive reference of EphrinB2 groups.
PCR product is done into detected through gel electrophoresis, the results are shown in Figure 4, wherein, swimming lane 1 is Marker, and swimming lane 2 is transfection
The recombinant HEK 293 cell of pEZ-M61-EphrinB2, swimming lane 3 are the HEK293 cells of untransfected, and swimming lane 4 is transfection empty carrier
HEK293 cells；
Compared with swimming lane 3,4, the band of 187bp that swimming lane 2 obtains or so is electric apparently higher than swimming lane 3,4, Ago-Gel
Swimming testing result shows the obvious up-regulation of EphrinB2 expression of recombinant HEK 293 cell.
D. under inverted fluorescence microscope in recombinant HEK 293 cell EphrinB2 expression and its and EphB4 acceptors combination
The expression of the EphrinB2 of lower observation recombinant HEK 293 cell under inverted fluorescence microscope, the results are shown in Figure 5,
Wherein a is control group HEK293 cells, and b is recombinant HEK 293 cell, and as can be seen from Figure 5 control group HEK293 cells are without green
Color fluorescence, and recombinant HEK 293 cell green fluorescence is brighter, illustrates that its EphrinB2 expression quantity dramatically increases.
Take growth conditions well close to the recombinant HEK 293 cell that converges, 0.25% Trypsin Induced, counts, by 1 ×
104A/hole concentration is inoculated in 96 well culture plates, per pore volume 200uL, is added after EphB4-Fc stostes (1mg/mL) are diluted
In 96 well culture plates, make its final concentration of 2.5ug/mL incubated cell, 24h is cultivated in 37 DEG C of incubators, due to recombinating HEK293
Cell itself carries green fluorescence, thus take pictures under the same visual field synthesising picture as shown in fig. 6, visible green with red overlap,
Illustrate the high expression EphrinB2 of recombinant HEK 293 cell and can preferably be combined with its ligand (acceptor) EphB4-Fc.
E.Western Blot detect the expression quantity of EphrinB2
With total protein of cell is extracted after lysate RIPA cell lysis, protein content is measured with BCA methods.Through electrophoresis, transferring film,
Immunostaining (primary antibody：Rabbit-anti people's EphrinB2 polyclonal antibodies, are purchased from Abcam companies；Secondary antibody：Goat antirabbit, is purchased from Bo Aosen
Company) and development, experimental result is as shown in fig. 7, wherein, 1 is recombinant HEK 293 cell, and 2 is thin for the HEK293 of transfection empty plasmid
Born of the same parents, 3 be the HEK293 cells of untransfected.Compared with 2,3, the expression quantity of target protein EphrinB2 is significantly raised in 1, in 2,3
The expression quantity of EphrinB2 is less；GAPDH groups are the positive reference of EphrinB2 groups.
F. the activity analysis of recombinant HEK 293 cell
Influence of the Dasatinib to restructuring HEK293 cell Proliferations is analyzed with mtt assay.
Take the logarithm the recombinant HEK 293 cell in growth period, the digestion of 0.25% membrane proteolytic enzyme, counts, by 1 × 104A/hole is dense
Degree is inoculated in 96 well culture plates, per pore volume 200uL, 37 DEG C, 5%CO2, cultivate 24h under the conditions of saturated humidity after, addition reaches
Sand replaces Buddhist nun solution 20uL, make Dasatinib final concentration be respectively 6.1,12.2,24.4,48.8,97.7,195.3,390.6mmol/
L, control group add equivalent drug solvent, each 5 multiple holes of concentration.Continue after cultivating 48h, careful inhale abandons nutrient solution in hole, often
Hole adds serum free medium 200uL, then adds 5mg/mL MTT solution 20uL, 37 DEG C of incubation 4h.Careful inhale abandons training in hole
Asked in nutrient solution, 150uLDMSO is added per hole, vibrated 10min, crystal is fully dissolved, measured with microplate reader under 490nm wavelength
Each hole absorbance, takes each multiple holes absorbance values, calculates inhibiting rate of the Dasatinib to cell according to the following formula：
Inhibiting rate (%)=(1- experimental groups absorbance/control group absorbance) × 100%
Using drug concentration as abscissa, inhibitory rate of cell growth is mapped for ordinate, and be inhibited effect curves, such as Fig. 8
It is shown：With the increase of Dasatinib concentration, Dasatinib is remarkably reinforced the inhibitory action of recombinant HEK 293 cell, through meter
Calculate, the half-inhibition concentration IC50 of Dasatinib is 107.9nmol/L；The EphrinB2 and Dasatinib of this explanation recombinant cell
With reference to rear, it is suppressed that the growth of the recombinant cell of EphrinB2 high expression.
In conclusion the recombinant HEK 293 cell of EphrinB2 high provided by the invention expression can be used for screening with
EphrinB2 is the medicine of target spot, so as to further obtain eliminating the medicine of influences of the EphrinB2 to tumor development.
Claims (2)
- A kind of 1. recombinant HEK 293 cell of EphrinB2 high expression, it is characterised in that：The recombinant cell is with HEK293 cells For host cell, exogenous expression's carrier of transfection host cell is the expression vector for including EphrinB2 full-length genes；The expression vector comprising EphrinB2 full-length genes is pEZ-M61-EphrinB2, wherein pEZ-M61- EphrinB2 is that EphrinB2 full-length genes are cloned into carrier pEZ-M61；Wherein, the EphrinB2 full-length genes Nucleotide sequence is as shown in SEQ.ID.NO.1；The primer sequence of the structure pEZ-M61-EphrinB2 is：Sense primer：5-TCATCTTCATCGTCATCATCATC-3；Anti-sense primer：5-CTGTCCGCAGTCCTTAGC-3.
- 2. the recombinant HEK 293 cell of the EphrinB2 high expression described in claim 1 is in the drug sieve using EphrinB2 as target spot The application chosen.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510094921.3A CN104694477B (en) | 2015-03-03 | 2015-03-03 | A kind of recombinant HEK 293 cell of EphrinB2 high expression and its application |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510094921.3A CN104694477B (en) | 2015-03-03 | 2015-03-03 | A kind of recombinant HEK 293 cell of EphrinB2 high expression and its application |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN104694477A CN104694477A (en) | 2015-06-10 |
| CN104694477B true CN104694477B (en) | 2018-04-17 |
Family
ID=53342035
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510094921.3A Expired - Fee Related CN104694477B (en) | 2015-03-03 | 2015-03-03 | A kind of recombinant HEK 293 cell of EphrinB2 high expression and its application |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104694477B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106047338B (en) * | 2016-06-21 | 2018-10-30 | 西安交通大学 | A kind of targeting EphrinB2 fluorescent tag molecule probes and its preparation method and application |
| CN110090295B (en) * | 2018-01-29 | 2022-12-23 | 武汉康理通科技有限公司 | Application of Ephrin-B1 in preparation of medicine for treating inflammatory bowel disease |
| CN114875070B (en) * | 2022-06-20 | 2024-05-24 | 西安交通大学 | GLP-1R expression vector, recombinant HEK293 cell based on same, construction method and application |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101875916A (en) * | 2009-11-27 | 2010-11-03 | 西安交通大学 | FGFR1 high-expression recombinant HEK293 cell and application thereof |
| CN103237812A (en) * | 2010-09-21 | 2013-08-07 | 国家肿瘤学研究中心基金会(Cnio) | Anti-ephrin-B2 antibody and use thereof |
| US8617883B2 (en) * | 2008-07-31 | 2013-12-31 | National University Of Ireland, Galway | Mesenchymal stem cell for promoting neovascularisation |
Family Cites Families (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPWO2007043629A1 (en) * | 2005-10-05 | 2009-04-16 | アキュメンバイオファーマ株式会社 | Method for inhibiting angiogenesis using ephrin B2 |
-
2015
- 2015-03-03 CN CN201510094921.3A patent/CN104694477B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US8617883B2 (en) * | 2008-07-31 | 2013-12-31 | National University Of Ireland, Galway | Mesenchymal stem cell for promoting neovascularisation |
| CN101875916A (en) * | 2009-11-27 | 2010-11-03 | 西安交通大学 | FGFR1 high-expression recombinant HEK293 cell and application thereof |
| CN103237812A (en) * | 2010-09-21 | 2013-08-07 | 国家肿瘤学研究中心基金会(Cnio) | Anti-ephrin-B2 antibody and use thereof |
Non-Patent Citations (2)
| Title |
|---|
| Accession NO:NM_004093,Homo sapiens ephrin-B2(EFNB2),mRNA;Nguyen TM,et al.;《Genbank》;20140511;FEATURES,ORIGIN部分 * |
| ephrinB2真核表达载体的构建及其在Hela细胞的表达;谢秋群 等;《湖南中医药大学学报》;20110228;第31卷(第2期);第3-5页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN104694477A (en) | 2015-06-10 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Liu et al. | TRPM7 channels regulate glioma stem cell through STAT3 and Notch signaling pathways | |
| Yao et al. | MSP–RON signalling in cancer: pathogenesis and therapeutic potential | |
| Lai et al. | Fibroblast activation protein regulates tumor-associated fibroblasts and epithelial ovarian cancer cells | |
| Hoque et al. | Aquaporin 1 is overexpressed in lung cancer and stimulates NIH-3T3 cell proliferation and anchorage-independent growth | |
| Jia et al. | SRp20 is a proto-oncogene critical for cell proliferation and tumor induction and maintenance | |
| Lee et al. | ELK3 promotes the migration and invasion of liver cancer stem cells by targeting HIF-1α | |
| Dong et al. | MicroRNA-335 suppresses the proliferation, migration, and invasion of breast cancer cells by targeting EphA4 | |
| Izquierdo et al. | Knockdown of T-cell intracellular antigens triggers cell proliferation, invasion and tumour growth | |
| Tsedensodnom et al. | Identification of T-cell factor-4 isoforms that contribute to the malignant phenotype of hepatocellular carcinoma cells | |
| Von Maltzahn et al. | A truncated Wnt7a retains full biological activity in skeletal muscle | |
| CN104694477B (en) | A kind of recombinant HEK 293 cell of EphrinB2 high expression and its application | |
| Scott et al. | Human papillomavirus type 16 E5-mediated upregulation of Met in human keratinocytes | |
| Ma et al. | Apoptosis-associated tyrosine kinase 1 inhibits growth and migration and promotes apoptosis in melanoma | |
| Feng et al. | MicroRNA-532-3p suppresses malignant behaviors of tongue squamous cell carcinoma via regulating CCR7 | |
| Huang et al. | FBXO31 promotes cell proliferation, metastasis and invasion in lung cancer | |
| Jia et al. | miR-375 inhibits cell growth and correlates with clinical outcomes in tongue squamous cell carcinoma | |
| Wu et al. | CIZ1 is upregulated in hepatocellular carcinoma and promotes the growth and migration of the cancer cells | |
| Vad-Nielsen et al. | Cause-and-Effect relationship between FGFR1 expression and epithelial-mesenchymal transition in EGFR-mutated non-small cell lung cancer cells | |
| Tian et al. | LncRNA LINC00974 downregulates miR-122 to upregulate RhoA in oral squamous cell carcinoma | |
| He et al. | NudCD1 promotes the proliferation and metastasis of non-small cell lung cancer cells through the activation of IGF1R-ERK1/2 | |
| CN101875916B (en) | FGFR1 high-expression recombinant HEK293 cell and application thereof | |
| CN101705227B (en) | SiRNA for inhibiting human AP-2alpha gene expression and anti-cervical cancer application thereof | |
| Indrigo et al. | Lentiviral vectors to study the differential function of ERK1 and ERK2 MAP kinases | |
| Furuhashi et al. | Ephrin receptor A4 expression enhances migration, invasion and neurotropism in pancreatic ductal adenocarcinoma cells | |
| Xu et al. | Adenovirus‑mediated overexpression of cystic fibrosis transmembrane conductance regulator enhances invasiveness and motility of serous ovarian cancer cells |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20180417 Termination date: 20210303 |