CN104698059B - A kind of glioma tumor markers and its application - Google Patents
A kind of glioma tumor markers and its application Download PDFInfo
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- 206010018338 Glioma Diseases 0.000 title claims abstract description 33
- 208000032612 Glial tumor Diseases 0.000 title claims abstract description 31
- 206010028980 Neoplasm Diseases 0.000 title claims abstract description 30
- 150000002270 gangliosides Chemical class 0.000 claims abstract description 24
- SQVRNKJHWKZAKO-UHFFFAOYSA-N beta-N-Acetyl-D-neuraminic acid Natural products CC(=O)NC1C(O)CC(O)(C(O)=O)OC1C(O)C(O)CO SQVRNKJHWKZAKO-UHFFFAOYSA-N 0.000 claims abstract description 6
- MSWZFWKMSRAUBD-GASJEMHNSA-N 2-amino-2-deoxy-D-galactopyranose Chemical compound N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O MSWZFWKMSRAUBD-GASJEMHNSA-N 0.000 claims abstract description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 3
- OVRNDRQMDRJTHS-KEWYIRBNSA-N N-acetyl-D-galactosamine Chemical compound CC(=O)N[C@H]1C(O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-KEWYIRBNSA-N 0.000 claims abstract description 3
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 claims abstract description 3
- SQVRNKJHWKZAKO-LUWBGTNYSA-N N-acetylneuraminic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)CC(O)(C(O)=O)O[C@H]1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-LUWBGTNYSA-N 0.000 claims abstract description 3
- MSWZFWKMSRAUBD-UHFFFAOYSA-N beta-D-galactosamine Natural products NC1C(O)OC(CO)C(O)C1O MSWZFWKMSRAUBD-UHFFFAOYSA-N 0.000 claims abstract description 3
- 239000008103 glucose Substances 0.000 claims abstract description 3
- 229940060155 neuac Drugs 0.000 claims abstract description 3
- CERZMXAJYMMUDR-UHFFFAOYSA-N neuraminic acid Natural products NC1C(O)CC(O)(C(O)=O)OC1C(O)C(O)CO CERZMXAJYMMUDR-UHFFFAOYSA-N 0.000 claims abstract description 3
- SQVRNKJHWKZAKO-OQPLDHBCSA-N sialic acid Chemical compound CC(=O)N[C@@H]1[C@@H](O)C[C@@](O)(C(O)=O)OC1[C@H](O)[C@H](O)CO SQVRNKJHWKZAKO-OQPLDHBCSA-N 0.000 claims abstract description 3
- -1 fatty acyl sphingosine Chemical compound 0.000 claims description 8
- 238000000034 method Methods 0.000 claims description 7
- 239000000203 mixture Substances 0.000 claims description 7
- WWUZIQQURGPMPG-UHFFFAOYSA-N (-)-D-erythro-Sphingosine Natural products CCCCCCCCCCCCCC=CC(O)C(N)CO WWUZIQQURGPMPG-UHFFFAOYSA-N 0.000 claims description 2
- 229930182470 glycoside Natural products 0.000 claims description 2
- 150000002338 glycosides Chemical group 0.000 claims description 2
- 238000004949 mass spectrometry Methods 0.000 claims 1
- 239000003814 drug Substances 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 3
- 238000002512 chemotherapy Methods 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 230000014509 gene expression Effects 0.000 description 3
- 238000001819 mass spectrum Methods 0.000 description 3
- 230000002018 overexpression Effects 0.000 description 3
- 238000011160 research Methods 0.000 description 3
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- 210000004881 tumor cell Anatomy 0.000 description 3
- 206010059866 Drug resistance Diseases 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 230000002378 acidificating effect Effects 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
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- 238000002474 experimental method Methods 0.000 description 2
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- BDERNNFJNOPAEC-UHFFFAOYSA-N propan-1-ol Chemical compound CCCO BDERNNFJNOPAEC-UHFFFAOYSA-N 0.000 description 2
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- 102100021569 Apoptosis regulator Bcl-2 Human genes 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
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- 101000971171 Homo sapiens Apoptosis regulator Bcl-2 Proteins 0.000 description 1
- 240000002853 Nelumbo nucifera Species 0.000 description 1
- 235000006508 Nelumbo nucifera Nutrition 0.000 description 1
- 235000006510 Nelumbo pentapetala Nutrition 0.000 description 1
- 229920005654 Sephadex Polymers 0.000 description 1
- 239000012507 Sephadex™ Substances 0.000 description 1
- 238000003639 Student–Newman–Keuls (SNK) method Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 239000000427 antigen Substances 0.000 description 1
- 102000036639 antigens Human genes 0.000 description 1
- 108091007433 antigens Proteins 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
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- 201000007980 brain meningioma Diseases 0.000 description 1
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- 238000005119 centrifugation Methods 0.000 description 1
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- 238000001959 radiotherapy Methods 0.000 description 1
- 125000005629 sialic acid group Chemical group 0.000 description 1
- YRTWNFXASSUQEW-UHFFFAOYSA-M sodium;methanol;acetate Chemical compound [Na+].OC.CC([O-])=O YRTWNFXASSUQEW-UHFFFAOYSA-M 0.000 description 1
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Abstract
The present invention relates to a kind of glioma tumor markers and its application.The glioma tumor markers is the gangliosides of double-sialylated, its structural formula any one specific as follows or multinomial shown：NeuAc‑α3‑Gal‑β3‑GalNAc‑β4（NeuAc‑α3）‑Gal‑β4‑Glu‑Cer；Gal‑β3‑GalNAc‑β4（NeuAc‑α8‑NeuAc‑α3）‑Gal‑β4‑Glu‑Cer；GalNAc‑β4（NeuAc‑α8‑NeuAc‑α3）‑Gal‑β4‑Glu‑Cer；NeuAc‑α8‑NeuAc‑α3‑Gal‑β4‑Glu‑Cer；Wherein, NeuAc represents sialic acid, and Gal represents galactolipin, and GalNAc represents galactosamine, and Glu represents glucose, and Cer represents N fatty acyl-sphingosines.The present invention can be used in the sign being expressly understood that of glioma tumour.
Description
Technical field
The present invention relates to tumor markers technical field, more particularly to a kind of glioma tumor markers and its application.
Background technology
Glioma is that occur one of tumour most commonly seen on mankind, and its deterioration degree is very high.It is general from making a definite diagnosis
Start, the life-span of patient is no more than 1 year.As one of tumour most commonly seen in central system disorder, glioma is accounted for into
The 30%-50% of people's intracranial tumors, the features such as having easily recurrence, treatment difficulty.The treatment means to glioma are still adopted at present
Based on operation, supplemented by radiotherapy chemotherapy.Due to the invasive growth of glioma, it can not be cut off completely during operation, so that also
It is that, after a ring mostly important in the auxiliary treatment after human glioma operation, but glioma is to all polyvoltine to determine chemotherapy
The drug resistance of medicine is treated, causes the therapeutic effect of the clinical chemotherapy of glioma undesirable.
Glycosyl sphingolipid is the class amphipathic molecule that a class is widely present on mammalian biological film, participates in maintaining cell
Normal physiological function.Research report in recent years shows, glycosyl sphingolipid can as tumour mark, it is or even also resistance to tumour
There is close relationship in the property of medicine.The accumulation of GlcCer and GCS in tumour cell can cause the resistance of tumour cell, therefore he
Same P-gp, Bcl-2 etc. be together considered as the tumour cell mark of tumor multi-medicine drug-resistant.Meanwhile, the synthesis precursor of glycosyl sphingolipid
It is GlcCer, exists in many tumours, such as breast cancer, oophoroma and epidermal carcinoma.Therefore, glycosyl sphingolipid and brain are further inquired into
The relation of glioma, the research of diagnoses and treatment and drug resistance for treating brain glioma disease has important help.
The content of the invention
The present inventor is it has been investigated that specificity overexpression is presented in the gangliosides of double-sialylated in glioma
Phenomenon, and there is no this specificity overexpression phenomenon in normal structure and other tumor tissues, therefore, it is possible to be used as brain colloid
The tumor marker of knurl, the present invention is completed based on above-mentioned discovery.
It is an object of the invention to provide a kind of glioma tumor markers, it is used as the significant of Diagnosing Gliomas
Material, characterizes the generation for the treatment of brain glioma disease in which can be expressly understood that.
The present invention includes herein below：
The present invention provides a kind of glioma tumor markers, and it is the gangliosides of double-sialylated.
In the present invention, the gangliosides of double-sialylated refer to the gangliosides for having two sialic acids in molecular formula.
As a preference of the present invention, the following any one of the structural formula of the gangliosides of the double-sialylated or multinomial institute
Show：
NeuAc-α3-Gal-β3-GalNAc-β4（NeuAc-α3）-Gal-β4-Glu-Cer；
Gal-β3-GalNAc-β4（NeuAc-α8-NeuAc-α3）-Gal-β4-Glu-Cer；
GalNAc-β4（NeuAc-α8-NeuAc-α3）-Gal-β4-Glu-Cer；
NeuAc-α8-NeuAc-α3-Gal-β4-Glu-Cer；
Wherein, NeuAc represents sialic acid, and Gal represents galactolipin, and GalNAc represents galactosamine, and Glu represents glucose,
Cer represents N fatty acyl sphingosine.
The gangliosides for the double-sialylated that structure above is represented are referred to as GD1a, GD1b, GD2 and GD3 successively.
As a preference of the present invention, the structure and/or composition of the gangliosides of the double-sialylated passes through mass spectrum side
Method is determined.
The present invention also provides above-mentioned glioma tumor markers in specificity junction mixture in connection is prepared
Using.
The specificity junction mixture, such as can be the monoclonal antibody or polyclonal antibody for resisting the tumor markers
Deng.The monoclonal antibody or polyclonal antibody can be that complete antibody or the part with binding site resist
Body fragment.
Preferably, the specificity junction mixture is the antibody for resisting the glioma tumor markers.That is, this hair
The antigen for the antibody that bright glioma tumor markers can resist the glioma tumor markers as preparing is used.
Beneficial effects of the present invention are：Research finds that the gangliosides of double-sialylated are presented specifically in glioma
Property high expression phenomenon, and there is no this specificity overexpression phenomenon in normal structure and other tumor tissues, therefore, it is possible to make
For the tumor markers of glioma, by the expression quantity of the gangliosides of Mass Spectrometer Method double-sialylated can quickly, it is accurate
Really and clearly determine the generation of glioma.
Brief description of the drawings
Fig. 1 is the first mass spectrometric figure of Main Gangliosides.
Fig. 2 is the structural formula schematic diagram of the gangliosides of double-sialylated.
Fig. 3 is expression of results figure of the gangliosides of double-sialylated in different tissues.
Embodiment
Embodiment of the present invention is described in detail below in conjunction with embodiment.Those skilled in the art will manage
Solution, following examples are only the preferred embodiments of the present invention, in order to more fully understand the present invention, thus should not be regarded as limiting this
The scope of invention.For those skilled in the art, the present invention can have various modifications and variations, all essences in the present invention
God is with principle, and any modification, equivalent substitution or improvement for being made etc. should be included in the scope of the protection.
Experimental method in following embodiments, is conventional method unless otherwise specified；Experiment material used, unless otherwise specified,
It is available from commercially available from routine biochemistry chemical reagent work.
Embodiment 1
The serum sample or tumor tissues sample of clinical Patients with gliomas are collected, tissue is ground or serum is mixed
It is placed in teat glass, chlorination is imitated：Methanol=1：1（v/v）Mixed solvent 2mL, gently smash tissue to pieces with dropper, ultrasonic 1h surpasses
Sound terminates, centrifugation（1500rpm, 5min）Supernatant is taken, extracting 4 times is repeated.Then transversion propyl alcohol：N-hexane：Water=55：25：20
（v/v/v）2mL extract solutions are extracted, and ultrasonic 1h, ultrasound terminates, centrifuging and taking supernatant, repeat extracting 4 times.Merge above-mentioned extracting supernatant
Liquid, is dried in rotary vacuum drier, and dried object places standby in -20 DEG C of low temperature refrigerator.Then sephadex A25 are crossed
Chromatographic column separating acid, neutral glycosphingolipid, with 0.8M sodium acetate methanol solution elution samples, it is to obtain acid sheath sugar to collect eluent
Fat component, equally collects 3 column volumes, is dried in rotary vacuum drier, and -20 DEG C of placements are standby after drying.By acidity
Glycosyl sphingolipid enters Mass Spectrometer Method, obtains the testing result of acidic glycosphingolipids as shown in figure 1, containing all main nerves in figure
Glycosides fat is saved, by further structural analysis, gangliosides can be divided into GM3, GM2, GM1, GD1, GD2 and GD3.Wherein
GD class gangliosides refer to the gangliosides of double-sialylated, its structure as shown in Fig. 2 the corresponding lotus matter in mass spectrum
Than as shown in table 1.
The charge-mass ratio result that the ganglioside lipodogramme of the double-sialylated of table 1 is obtained
Embodiment 2
5 normal cerebral tissue's samples that attached First People's Hospital of University Of Suzhou is obtained（Control）, 6 brain meningioma tissues
Sample, 16 hypophysoma samples and 14 glioma samples, extract glycosyl sphingolipid, and carry out the mass spectrum of acidic glycosphingolipids respectively
Analysis（Such as embodiment 1）.The relative abundance of total gangliosides is designated as M（Always）, tri- kinds of double-sialylateds of GD1, GD2, GD3
The relative abundances of gangliosides be denoted as M（GD）, then the content of the gangliosides of double-sialylated in the tissue
It is expressed as R=M（GD）/M（Always）*100%.R values in every group of sample are carried out into Student-Newman-Keuls inspections to be united
Credit analysis, obtained result are counted as shown in figure 3, the level of the gangliosides of double-sialylated is obvious high in glioma
In normal control and hypophysoma group and meningioma group, this illustrates the gangliosides double-sialylated as glioma
Tumor markers is feasible.
Applicant states that the present invention illustrates the detailed features and method detailed of the present invention by above-described embodiment, but
The invention is not limited in above-mentioned detailed features and method detailed, that is, do not mean that the present invention has to rely on above-mentioned detailed features
And method detailed could be implemented.Person of ordinary skill in the field is it will be clearly understood that any improvement in the present invention, to this hair
The equivalence replacement of bright selection component and the addition of auxiliary element, the selection of concrete mode etc., all fall within protection scope of the present invention
Within the scope of disclosure.
Claims (4)
1. a kind of glioma tumor markers, it is the gangliosides of double-sialylated；
The following any one of the structural formula of the gangliosides of the double-sialylated is multinomial shown：
NeuAc-α3-Gal-β3-GalNAc-β4(NeuAc-α3)-Gal-β4-Glu-Cer；
Gal-β3-GalNAc-β4(NeuAc-α8-NeuAc-α3)-Gal-β4-Glu-Cer；
GalNAc-β4(NeuAc-α8-NeuAc-α3)-Gal-β4-Glu-Cer；
NeuAc-α8-NeuAc-α3-Gal-β4-Glu-Cer；
Wherein, NeuAc represents sialic acid, and Gal represents galactolipin, and GalNAc represents galactosamine, and Glu represents glucose, Cer generations
Table N fatty acyl sphingosine.
2. glioma tumor markers according to claim 1, it is characterised in that the neuromere of the double-sialylated
The structure and/or composition of glycosides fat is determined by mass spectrometry method.
3. the glioma tumor markers as described in claim any one of 1-2 is preparing specificity junction mixture in connection
In application.
4. application according to claim 3, it is characterised in that the specificity junction mixture is to resist the glioma tumour
The antibody of mark.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2172219A1 (en) * | 2008-10-02 | 2010-04-07 | SNU R&DB Foundation | Anti-cancer agent comprising an iNKT ligand and anti-PD-1 antibody or anti-PD-L1 antibody |
| CN101809148A (en) * | 2007-07-27 | 2010-08-18 | 伊玛提克斯生物技术有限公司 | Novel immunotherapy of neuronal and brain tumors |
| CN102095845A (en) * | 2009-12-11 | 2011-06-15 | 上海裕隆生物科技有限公司 | Chemiluminescence quantitative detection kit for carbohydrate antigen 19-9 |
| CN102170900A (en) * | 2008-10-01 | 2011-08-31 | 伊玛提克斯生物技术有限公司 | Novel immunotherapy against several tumors, including neuronal and brain tumors |
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2013
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Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101809148A (en) * | 2007-07-27 | 2010-08-18 | 伊玛提克斯生物技术有限公司 | Novel immunotherapy of neuronal and brain tumors |
| CN102170900A (en) * | 2008-10-01 | 2011-08-31 | 伊玛提克斯生物技术有限公司 | Novel immunotherapy against several tumors, including neuronal and brain tumors |
| EP2172219A1 (en) * | 2008-10-02 | 2010-04-07 | SNU R&DB Foundation | Anti-cancer agent comprising an iNKT ligand and anti-PD-1 antibody or anti-PD-L1 antibody |
| CN102095845A (en) * | 2009-12-11 | 2011-06-15 | 上海裕隆生物科技有限公司 | Chemiluminescence quantitative detection kit for carbohydrate antigen 19-9 |
Non-Patent Citations (1)
| Title |
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| 《C18鞘糖脂在脑胶质瘤中的表达及其在耐药中的功能研究》;俞云会;《中国优秀硕士论文全文数据库》;20130315;正文第19页 * |
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