CN104558146A - Molecular target Gab2 and application thereof in preparing medicines and health care food for preventing and treating fatty liver and liver cancer - Google Patents
Molecular target Gab2 and application thereof in preparing medicines and health care food for preventing and treating fatty liver and liver cancer Download PDFInfo
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Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
- C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
-
- 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
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention discloses molecular target Gab2 and application thereof in preparing medicines and health care food for preventing and treating fatty liver and liver cancer, and relates to a molecular target. The molecular target Gab2 is cytoplasm signal scaffolding protein mediating pathogenic factor-induced fatty liver and liver cancer; multiple disease animal models prove that in fatty liver and related diseases, the expression of the Gab2 can be induced by pathogenic factor; by collecting and integrating various signals by virtue of the Gab2, a comprehensive effect is generated, so that the development of diseases is promoted. The molecular target Gab2 can be applied to prepare medicines for preventing and treating fatty liver and liver cancer, and the molecular target Gab2 can be also used for preparing health care food for preventing and treating fatty liver and liver cancer.
Description
Technical field
The present invention relates to molecular target, especially relate to molecular target Gab2 and preparing the application in prevention and therapy fatty liver and liver-cancer medicine and health food.
Background technology
Non-alcohol and alcoholic fatty liver are that society threatens one of principal disease of human health, and the incidence of China is particularly serious.Improving constantly particularly in recent years along with people's living standard, dietary structure and life style have larger change, and fatty liver etc. " rich man's disease " incidence of disease is more and more higher.No matter be alcoholic fatty liver or NASH, all relating to a complicated mechanism, is all the pathologic effect that multiple adjustment signal comprehensively produces.The regulation and control of this complexity and various adjustment signal, the treatment for disease brings very large obstacle, causes the treatment carried out for single or a small amount of signal, effect not ideal enough [1,2].Therefore, people expect the key node that can find signals-modulating in fatty liver in recent years, particularly disclosing those can the platform albumen of integrated signal, albumen as similar in IRS, by changing these key proteins, change the comprehensive organism effect of signal, thus disease therapy [1-3].Gab (Grb2binding protein) family protein, comprises mammiferous Gab1, Gab2, Gab3, may be exactly the such albumen of a class.
Gab family protein as the scaffolding protein of a signal, in conjunction with cell membrane, raise the path in various kinds of cell, can amplify, integrate the signal coming from acceptor, be considered to the hinge [4] of signals-modulating.Interaction can be there is in vivo in Gab2 with many downstream signaling molecules, such as AKT1 [5], CRKL [5], Grb2, PI3K, PTPN11 (shp2) [6] and PLCG2 equimolecular, regulating cell propagation, survival, migration and the function [7,8] of breaking up.There is unconventionality expression phenomenon in Gab2, such as, all play very important effect [4] at leukaemia, oophoroma, Alzheimer disease in numerous disease.At Liver Lipid Metabolism, the Effect study of Gab2 is little.But Gab1 knocks out Gab1 to increase mouse sugar tolerance and to improve mouse islets element active [9] in liver.In addition, SHP2 plays a role in vivo and is normally combined with Gab2, and then the signal transduction in regulation and control downstream.A lot of evidence shows that Shp2 also can by the sugared stable state [10-12] in the metabolism of the regulation and control degraded of fatty acid synthase and leptin path regulation and control liver lipids and body.But at present for the effect of Gab2 in lipid-metabolism and metabolic disease, need deep research.
List of references
1.Birkenfeld,A.L.,et al.(2014)Nonalcoholic fatty liver disease,hepaticinsulin resistance,and type 2diabetes.Hepatology,59,713-23.
2.Farrell,G.C.,et al.(2013)NAFLD in Asia--as common and important as in theWest.Nat Rev Gastroenterol Hepatol,10,307-18.
3.White,M.F.(2014)IRS2integrates insulin/IGF1signalling with metabolism,neurodegeneration and longevity.Diabetes Obes Metab,16Suppl 1,4-15.
4.Nakaoka,Y.,et al.(2013)Gab docking proteins in cardiovascular disease,cancer,and inflammation.Int J Inflam,2013,141068.
5.Pan,X.L.,et al.(2010)The Gab2in signal transduction and its potentialrole in the pathogenesis of Alzheimer's disease.Neurosci Bull,26,241-6.
6.Million,R.P.,et al.(2004)A direct binding site for Grb2contributes totransformation and leukemogenesis by the Tel-Abl(ETV6-Abl)tyrosine kinase.Mol CellBiol,24,4685-95.
7.Yan Liu,L.R.R.(2002)The gift of Gab.FEBS Lett,515,1-7.
8.Simister,P.C.,et al.(2012)Order and disorder in large multi-site dockingproteins of the Gab family--implications for signalling complex formation andinhibitor design strategies.Mol Biosyst,8,33-46.
9.Bard-Chapeau,E.A.,et al.(2005)Deletion of Gab1in the liver leads toenhanced glucose tolerance and improved hepatic insulin action.Nat Med,11,567-71.
10.Zhang,E.E.,et al.(2004)Neuronal Shp2tyrosine phosphatase controlsenergy balance and metabolism.Proc Natl Acad Sci U S A,101,16064-9.
11.Yu,J.,et al.(2013)Modulation of fatty acid synthase degradation byconcerted action of p38MAP kinase,E3ligase COP1,and SH2-tyrosine phosphatase Shp2.J Biol Chem,288,3823-30.
12.He,Z.,et al.(2012)Shp2controls female body weight and energy balanceby integrating leptin and estrogen signals.Mol Cell Biol,32,1867-78.
Summary of the invention
The first object of the present invention is to provide the molecular target Gab2 controlling fatty liver and liver cancer.
The second object of the present invention is to provide molecular target Gab2 preparing the application in prevention and therapy fatty liver and liver-cancer medicine.
The third object of the present invention is to provide molecular target Gab2 preparing the application in prevention and therapy fatty liver and liver cancer health food.
Described molecular target Gab2 (Grb2binding protein 2) a kind ofly mediates the tenuigenin signal support albumen that paathogenic factor brings out fatty liver and liver cancer; Utilize various diseases animal model, prove in fatty liver and relevant disease, virulence factor induction Gab2 expresses, and is raised and integrates multi-signal, produce a comprehensive effect by Gab2, and leading advancing of disease.
Proved by 124 routine people's liver cancer sample census, Gab2 high expressed is in most of case; The mouse of Gab2 gene knockout, can resist the obesity of high lipid food induction and the accumulation of fat in liver, alcohol produces liver acute or chronic injury and fat, and the fatty hepatitis of MCD induction.Experiment of nude mouse proves, reduces Gab2 protein level in hepatoma cell line, and also blocks cellular is in the one-tenth knurl of nude mice.Therefore, can prevention and therapy fatty liver and relevant hepatitis and liver cancer by reducing that Gab2 expresses, utilize the expression detecting Gab2 albumen can diagnose these diseases.
As can be seen here, molecular target Gab2 can apply in the medicine preparing prevention and therapy fatty liver and liver cancer, and molecular target Gab2 also can apply in the health food preparing prevention and therapy fatty liver and liver cancer.
Accompanying drawing explanation
Fig. 1 is Gab2 albumen high expressed in NASH (NAFLD) murine liver tissue.
Fig. 2 is Gab2 albumen high expressed in chronic alcoholic fatty liver (NAFLD) murine liver tissue
Fig. 3 a is the expression of Gab2 albumen in fresh HCC sample.Protein level antibody test, T marked tumor tissue, P is his cancer beside organism.
Fig. 3 b is the quantitative statistics analysis of protein expression in fresh HCC sample.**P<0.01。
Fig. 4 a is the figure of non-alcohol liver disease model mouse (high lipid food of feeding).
Fig. 4 b is the quantitative statistics analysis of non-alcohol liver disease model mouse (high lipid food of feeding).^^p<0.01,*p<0.05。
Fig. 5 a is the liver of non-alcohol liver disease model mouse (high lipid food of feeding).
Fig. 5 b is the liver of non-alcohol liver disease model mouse (high lipid food of feeding) and the ratio quantitative statistics of body weight.^^,**p<0.01。
Fig. 6 a is the histotomy of the liver of non-alcohol liver disease model mouse (high lipid food of feeding).HE dyes display, and cavity is that mixed type fat drips.
Fig. 6 b is the quantitative statistics that the histotomy fat of the liver of non-alcohol liver disease model mouse (high lipid food of feeding) drips cavity area.^^p<0.01,*p<0.05。
Fig. 7 is the level of the triglyceride (TG) of non-alcohol liver disease model mouse (high lipid food of feeding) serum.^^,**p<0.01。
Fig. 8 is the level of the T-CHOL (T-CHO) of non-alcohol liver disease model mouse (high lipid food of feeding) serum.^^,**p<0.01。
Fig. 9 is the liver organization of acute alcohol liver model mice.
Figure 10 is the liver organization of Chronic Alcohol liver model mice.
Figure 11 is the concentration of Chronic Alcohol liver model mice Serum ALT (glutamic-oxalacetic transaminease).^^,**p<0.01。
Figure 12 is the concentration of Chronic Alcohol liver model mice serum triglyceride.^^,**p<0.01。
Figure 13 is the liver organization (HE dyeing) of NASH model mice.* mark fat and drip cavity, arrow points inflammatory cell infiltration region.
Figure 14 is horizontal .^^, the * * p<0.01 of the triglyceride (TG) of NASH mice serum.
Figure 15 is horizontal .^^, the * * p<0.0 of the T-CHOL (T-CHO) of NASH mice serum.
Figure 16 is horizontal .^^, the * * p<0.0 of the TNF α of NASH mice serum.
Figure 17 a be Gab2 process LAN or strike the HepG2 hepatoma carcinoma cell of falling tie up to nude mice one-tenth knurl experiment in small animal living body imaging shooting in body tumor shape exemplary plot.
Figure 17 b is Gab2 process LAN or strikes the fast statistical study of knurl that the HepG2 hepatoma carcinoma cell of falling ties up to all animals in the one-tenth knurl experiment of nude mice.
Figure 18 a is the cell growth curve that Gab2 process LAN or the propagation mtt assay striking the HepG2 cell fallen detect.**P<0.01。
Figure 18 b is the Gab2 protein expression of cell in Gab2 process LAN or the propagation of striking the HepG2 cell fallen.
Figure 19 a is Gab2 process LAN or the migration example figure striking the HepG2 cell fallen.
Figure 19 b is the quantitative statistics analysis of Gab2 process LAN or the migration test of many times of striking the HepG2 cell fallen.**P<0.01。
Embodiment
The mouse liver that embodiment 1:Gab2 high expressed is fed in high lipid food or alcohol, and the liver cancer case of the people of more than 60%, can as the mark of diagnosing cancer of liver.
(1). mouse nonalcoholic fatty liver model
Screening Gab2 (entirely strike/partly strike) mouse carries out NASH modeling experiment.Experiment choose 12 8 week age Gab2 (-/-) male mice and 12 8 week age Gab2 (+/-) male mice, often kind of mouse is divided into two groups at random, often organize 6, be respectively control group and high fat group, control group fed normal mouse basestocks, high fat group is fed high lipid food (basal feed 77.9%+ lard 10%, cholesterol 2%, yolk powder 10%, cholate 0.1%).Each group starts regularly to weigh in afterwards respectively at modeling, for drafting growth curve, and when modeling 6 months Mouse Weight no longer include rise appreciably after put to death, take out complete liver and carry out the expression that western blot tests detection Gab2 albumen, find Gab2 albumen high expressed (as Fig. 1) in NASH (NAFLD) mouse model.
(2) in order to verify the relation between Gab2 and Liver Lipid Metabolism, simultaneously also with Lieber-DeCarli containing spirituous liquid feed and etc. the mouse 6 weeks of heat isopyknic control group feed feeding C57BL/6, change freshly prepared liquid feed when every morning 9.Shift out liver and carry out immunoblot experiment checking Gab2 expression change.The expression showed increased (as Fig. 2) of Gab2 in result display alcohol feeding group mouse liver.
(3) people's liver cancer sample
Tissue homogenate instrument is utilized to be extracted 60 routine 0.1cm
3freshman hepatic tissue (30 routine liver cancer, the other contrast of 30 routine cancers) albumen and have detected Gab2 protein expression, result display wherein has the other sample Gab2 of 21 liver cancer tissues its cancer relative to there is high expressed in various degree, and result is as Fig. 3 a, and the quantitative statistics analysis of protein expression is see Fig. 3 b.
In addition, the Gab2 detecting 94 routine liver cancer tissues and 23 normal structures expresses, and find most of liver cancer case Gab2 high expressed, and normal structure only has a small amount of expression, in protein chip in liver cancer sample the expression statistical study of Gab2 albumen as table 1.
Table 1
-，+，++，+++represent different degrees of staining
In tissue, protein level SABC detects, and determines according to the degree of the positive and the proportion composite of positive cell the different brackets that Gab2 expresses, with+number to represent.
Mouse obesity and the liver fat of the induction of embodiment 2:Gab2 gene knockout retardance high lipid food are piled up.
Start regularly to weigh in afterwards in NAFLD modeling, and when modeling 6 months Mouse Weight no longer include rise appreciably after put to death, take pictures, weigh in, liver is heavy, epididymal adipose tissues weight, blood sampling detects serum paddy propionamide transaminase (ALT), triglyceride (TG), T-CHOL (TC), takes out after complete liver HE dyes and does pathological observation.
1, Gab2 gene to knock out the mouse effectively reducing the induction of high fat fat
When modeling carries out 24 weeks, mouse weights, all there is fat phenomenon in the contrast separately of Gab2 (knocking out/heterozygosis) mouse height fat group, body weight obviously increases, and the high fat mouse obese degree that Gab2 knocks out group is comparatively light, body weight comparatively heterozygosis group mouse also significantly decreases (as Fig. 4 a and Fig. 4 b).
2, Gab2 gene knock out effective hepar damnification and the lipid accumulation situation of improving the induction of high fat
Visual inspection, blank group mouse liver is scarlet, clear-cut margin, matter is tough, Models of Fatty Liver group liver volume increases in tawny, edge rust, and greasy feeling increases the weight of, and relative to heterozygosis mouse, knocking out of Gab2 gene makes experimental mice liver index and morphology lesion degree weaken (as Fig. 5 a and Fig. 5 b).
Light microscopic HE dyes display, and Gab2 knocks out with the control group liver cell of Gab2 heterozygosis group all in polygon, core circle, and between two parties, endochylema enriches, and eucaryotic cell structure is clear, without other pathological changes.The hyperlipidemia model group liver cell obvious tumefaction of Gab2 chimeric mice, hepatic cell fattydegeneration is obvious, exist in kytoplasm and drip based on the mixed type fat of large alveolitoid, cavity pushes nucleus to side; pathology involves whole lobuli hepatis; the large fat filling the air distribution in hepatic tissue as seen drips and drips with micro-fat, and scatters full liver; Comparatively speaking, Gab2 knocks out the hyperlipidemia model group of group, and pathology situation is obviously comparatively light, and according to quantitative statistics, the quantity that fat drips and size all improve significantly (as Fig. 6 a and Fig. 6 b).
Meanwhile, determine the level of triglyceride (TG) (as Fig. 7) and T-CHOL (T-CHO) (as Fig. 8) of mice serum, Gab2 knocks out the expression (as Fig. 8) reducing the two equally.
In sum, that can see Gab2 gene knocks out the hepar damnification degree and lipid accumulation situation that improve to a certain extent and brought by the nursing of high lipid food.
The mouse of embodiment 3:Gab2 gene knockout, can resist alcohol and produce liver acute or chronic injury and fat.
In addition, equally find that or chronic injury acute at the mouse liver of alcohol induction and fat produce and be also suppressed because of knocking out of Gab2 gene.First construct acute with chronic alcoholic Models of Fatty Liver.Acute model adopts fills with food alcohol (6g/kg body weight) and the experimental technique with the isocaloric maltodextrin of alcohol equal-volume, adult mice is divided into four groups at random according to genotype, fill with food alcohol and contrast liquid respectively, after 12 hours, get blood plasma and liver.The formation (as Fig. 9) that the fat significantly improving alcohol induction after result display knocks out the mouse feeding alcohol of Gab2 gene drips.
The structure of chronic alcoholic Models of Fatty Liver is according to the method for list of references, adult mice is divided into four groups (n=6/ groups) at random according to genotype, respectively feeding Lieber-DeCarli containing spirituous liquid feed and etc. the isopyknic control group feed of heat, every morning 9 changes freshly prepared liquid feed, and feeding was drawn materials after 6 weeks.The packing phenomenon (as Figure 10) that the fat that the knocking out of same discovery Gab2 gene significantly suppress alcohol feed induction at a slow speed drips, and the rise phenomenon of mice serum ALT (glutamic-oxalacetic transaminease) (as Figure 11), triglyceride (as Figure 12) also obviously improves (as Figure 11).These change describe Gab2 knock out the generation can resisting acute or chronic liver injury that alcohol causes and lipid.
The mouse of embodiment 4, Gab2 gene knockout, can resist the fatty hepatitis of choline methionine deficiency (MCD) model induction.
Utilize classical nonalcoholic steatohepatitis (NASH) modeling method, methionine-choline lacks model (MCD), and carries out mouse modeling in contrast with methionine-choline interpolation feed (MCS).Experiment choose 12 8 week age Gab2 (-/-) male mice and 12 8 week age Gab2 (+/-) male mice, often kind of mouse is divided into two groups at random, often organize 6, be respectively MCD group and MCS group, experiment is carried out mouse weights after 8 weeks, gets liver, is got blood.Carry out the HE dyeing of hepatic tissue subsequently respectively, and measure Triglycerides in Serum, T-CHOL and inflammatory factor TNF alpha content respectively.Found that, Gab2 gene knock out the damage effectively can alleviated fatty liver inflammation and liver is brought, can see from HE coloration result, the liver cell fat vacuole of MCD induction and inflammatory cell region are obtained in Gab2-/-mouse improves (as Figure 13) significantly, and the rising of the serum TNF α that NASH mice serum triglyceride (TG) (as Figure 14), T-CHOL (T-CHO) (as Figure 15) and inflammation-induced cause is also because knocking out of Gab2 gene significantly reduces (as Figure 16).
Embodiment 5, in hepatoma cell line, reduce Gab2 protein level, stop the growth of cell, migration and the one-tenth knurl in nude mouse.
The research that Gab2 acts in liver cancer verifies by being tied to form knurl to mouse bare subcutaneous injection hepatoma carcinoma cell.Nude mice (5 week age) is bought on behalf by Xiamen University's animal experimental center, male and female half and half.Be put in Xiamen University animal experimental center standard SPF level free choice feeding one week, then carry out hypodermic injection hepatoma cell line.Four kinds of clone is respectively EGFP cellular control unit, EGFP-Gab2 process LAN Gab2 group cell, EGFP-Gab2 turns control plasmid group cell and EGFP-Gab2 turns Gab2-siRNA group cell.Often organize cell infusion 1x10
7individual cell/only, raise 30 days under normal condition after injection.Finally, carry out tumor size measurement with vernier caliper, and with small animal living body imaging shooting at body tumor shape.Result proves, Gab2 protein decreased suppresses the one-tenth knurl of HepG2 cell in nude mice figure (as Figure 17 a and Figure 17 b).
Nude mice becomes knurl modeling simultaneously, also reduce the impact on liver cancer cell growth after Gab2 expresses with MTT experiment checking, respectively five kinds of clones are inoculated in 96 orifice plates with 2000 cells/well, the light absorption value of cell and 492nm wavelength is detected respectively at the 1st, 2,3 and 4 day, finally add up mapping analysis and find that process LAN Gab2 clone obviously facilitates the growth of tumour cell, but but significantly suppress the propagation (as Figure 18 a and Figure 18 b) of tumour cell after endogenous cellular Gab2 albumen is reduced expression.
For tumor migration experiment, by 1x10
5it is in the Transwell of 8 μm that individual cell is inoculated in pore size, the upper room of Transwell adds serum free medium, lower room adds the complete medium containing 10% hyclone, cultivate after 12 hours and upper room film superficial cell is wiped, then film surface in lower room is carried out to the violet staining analysis of 0.2%.By coloration result with 33% glacial acetic acid elute, and detect the difference of light absorption value under 492nm wavelength.Detect equally after reducing Gab2 expression, obviously inhibit the excessive transport phenomena (as Figure 19 a and Figure 19 b) of EGFP-Gab2 clone.
Claims (3)
1. molecular target Gab2, it is characterized in that a kind ofly mediating the tenuigenin signal support albumen that paathogenic factor brings out fatty liver and liver cancer.
2. molecular target Gab2 applies in the medicine preparing prevention and therapy fatty liver and liver cancer as claimed in claim 1.
3. molecular target Gab2 applies in the health food preparing prevention and therapy fatty liver and liver cancer as claimed in claim 1.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106577476A (en) * | 2016-11-02 | 2017-04-26 | 唐博 | Method for establishing hepatic fibrosis model through hyperhomocysteinemia induction |
| CN114015684A (en) * | 2021-08-27 | 2022-02-08 | 南通市肿瘤医院 | GAB2 small interfering RNA fragment and application thereof |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20080081791A1 (en) * | 2006-07-06 | 2008-04-03 | Weida Huang | Methods of using combinations of siRNAs for treating a disease or a disorder, and for enhancing siRNA efficacy in RNAi |
| US20100130592A1 (en) * | 2002-02-20 | 2010-05-27 | Sirna Therapeutics, Inc. | RNA Interference Mediated Inhibition of GRB2 Associated Binding Protein (GAB2) Gene Expression Using Short Interfering Nucleic Acid (siNA) |
-
2015
- 2015-01-20 CN CN201510026681.3A patent/CN104558146A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20100130592A1 (en) * | 2002-02-20 | 2010-05-27 | Sirna Therapeutics, Inc. | RNA Interference Mediated Inhibition of GRB2 Associated Binding Protein (GAB2) Gene Expression Using Short Interfering Nucleic Acid (siNA) |
| US20080081791A1 (en) * | 2006-07-06 | 2008-04-03 | Weida Huang | Methods of using combinations of siRNAs for treating a disease or a disorder, and for enhancing siRNA efficacy in RNAi |
Non-Patent Citations (6)
| Title |
|---|
| ADAMS等: "GAB2—a Scaffolding Protein in Cancer", 《MOLECULAR CANCER RESEARCH》 * |
| MALAGUARNERA等: "Molecular mechanisms involved in NAFLD progression", 《J MOL MED(BERL)》 * |
| MATSUMURA等: "Clinical significance of GAB2, a scaffolding/docking protein acting downstream of EGFR in human colorectal cancer.", 《ANN SURG ONCOL》 * |
| PUROHIT等: "Molecular Mechanisms of Alcoholic Fatty Liver", 《ALCOHOL CLIN EXP RES》 * |
| 中华人民共和国国家卫生和计划生育委员会: "《食品安全国家标准 保健食品》", 24 December 2014 * |
| 吕忠显: "《中国细胞生物学学会全体会员代表大会暨第十二次学术大会论文摘要集》", 16 July 2011 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106577476A (en) * | 2016-11-02 | 2017-04-26 | 唐博 | Method for establishing hepatic fibrosis model through hyperhomocysteinemia induction |
| CN106577476B (en) * | 2016-11-02 | 2021-01-15 | 唐博 | Method for establishing hyperhomocysteinemia induced liver fibrosis model |
| CN114015684A (en) * | 2021-08-27 | 2022-02-08 | 南通市肿瘤医院 | GAB2 small interfering RNA fragment and application thereof |
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