CN107884491A - The combination of stomach cancer amino acid tags thing, screening technique and its application in diagnosing gastric cancer - Google Patents
The combination of stomach cancer amino acid tags thing, screening technique and its application in diagnosing gastric cancer Download PDFInfo
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- CN107884491A CN107884491A CN201711087774.2A CN201711087774A CN107884491A CN 107884491 A CN107884491 A CN 107884491A CN 201711087774 A CN201711087774 A CN 201711087774A CN 107884491 A CN107884491 A CN 107884491A
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Abstract
The present invention relates to the diagnostic techniques field of stomach cancer, disclose a kind of combination of stomach cancer amino acid tags thing, screening technique and its application in diagnosing gastric cancer, the metabolism amino acid in patients with gastric cancer and Healthy People saliva sample is determined using high performance liquid chromatography GC-MS, and filters out the metabolism amino acid related with stomach cancer and is combined as stomach cancer amino acid tags thing；The stomach cancer amino acid tags thing combination includes following several or whole combinations：Taurine, glycine, glutamine, monoethanolamine, histidine, alanine, glutamic acid, hydroxylysine, proline and tyrosine.Compared with prior art, after the combination of stomach cancer amino acid tags thing is applied in diagnosing gastric cancer in the present invention, it can not only quickly and easily detect whether testee suffers from stomach cancer, the progressive stage of stomach cancer can also be determined, noninvasive close friend is diagnosed, simple, convenient and rapid, accuracy rate is high, be advantageous to determine clinical treatment, improve cure rate.
Description
Technical field
The present invention relates to the diagnostic techniques field of stomach cancer, more particularly to a kind of stomach cancer amino acid tags thing combination, screening side
Method and its application in diagnosing gastric cancer.
Background technology
Stomach cancer is one of current most common malignant cancer, and it comes all cancers respectively in the morbidity and mortality of China
Second in disease and the 3rd.At present, the best approach of curing gastric cancer is only early diagnosed.Statistics shows early carcinoma of stomach
Postoperative 5 years survival rates are up to 90%, and late gastric cancer is less than 24%.However, 80% stomach cancer symptom in early stage is very not clear
It is aobvious, easily it is ignored or fails to pinpoint a disease in diagnosis.When that clinically can obtain making a definite diagnosis, stomach cancer has often had evolved to middle and advanced stage, causes patient
Miss optimal therapic opportunity.With the popularization of gastrocopy, the early diagnostic rate of stomach cancer has obtained certain lifting, still
Gastrocopy is invasive, and patient is more painful in checking process, and expensive, and simple gastrocopy can not carry out stomach cancer point
Phase, it is necessary to which by biopsy or pathologic examination after operation, process is longer, and these reasons cause it can not turn into the reason of stomach cancer Mass screening
Think method.Therefore, the method that cancer is most worthy is early diagnosed by detecting particular cancers Research of predicting markers in biological specimen.
The biological specimen includes various body fluid.For many main cancers, in body fluid there is spirit in protide mark
The problem of sensitivity and insufficient specificity.For example, for protide mark cal9-9 the most frequently used in patients with gastric cancer body fluid,
Ca72-4 and carcinomebryonic antigen (CEA) can only detect the stomach neoplasm of about 15-50% either phase, for early stage disease, be down to
About 2-11%.And in various body fluid, saliva can be acquired non-invasively, it is convenient, accurate and without any ethics and
Medical science risk.Secondly, several amino acids contained in saliva and stomach cancer have it is close associate, these amino acid classes are numerous
And chemical property is stable, the difference for reflecting early carcinoma of stomach and late gastric cancer can be used for as reliable stomach cancer marker, therefore
Interindividual variation can be at utmost eliminated, is suitable for establishing a kind of general and reliable stomach cancer diagnostic method, quick experiment
The early diagnosis of existing stomach cancer.
It there is no and stomach cancer diagnosed as mark at present using the amino acid in saliva.If can timely diagnosis of gastric cancer
In the residing stage, clinical treatment will be favorably determined, will be suited the remedy to the case, and improve cure rate.However, existing diagnosed using saliva
Stomach cancer method also can not effectively be distinguished to early and late stomach cancer.
The content of the invention
Goal of the invention：For problems of the prior art, a kind of stomach cancer amino acid tags thing combination of present invention offer,
Screening technique and its application in diagnosing gastric cancer, the present invention in stomach cancer amino acid tags thing can be used for by human saliva
Detection quick diagnosis go out whether testee suffers from stomach cancer, and can determine stomach cancer virus developing stage, diagnose noninvasive friend
Good, simple, convenient and rapid, accuracy rate is high, is advantageous to determine clinical treatment, improves cure rate.
Technical scheme：The invention provides a kind of combination of stomach cancer amino acid tags thing, including following several or whole groups
Close：Taurine, glycine, glutamine, monoethanolamine, histidine, alanine, glutamic acid, hydroxylysine, proline and junket ammonia
Acid.
Present invention also offers a kind of screening technique of stomach cancer amino acid tags thing combination, high performance liquid chromatography-matter is utilized
The metabolism amino acid in combined instrument measure patients with gastric cancer and Healthy People saliva sample is composed, and filters out the metabolism ammonia related to stomach cancer
Base acid combines as stomach cancer amino acid tags thing；The stomach cancer amino acid tags thing combination includes following several or whole groups
Close：Taurine, glycine, glutamine, monoethanolamine, histidine, alanine, glutamic acid, hydroxylysine, proline and junket ammonia
Acid.
Preferably, the condition of the high performance liquid chromatography is：From C18 chromatographic columns, water and 2% that Flow sample is 98%
Acetonitrile, it is percent by volume, carries out gradient activation；Flow velocity is 0.8mL/min, and column temperature is room temperature.The initial equilibration time is
5min, analysis run time are 25min, are carried out under room temperature condition.
Preferably, the gradient activation is arbitrarily carried out according to following condition：(1) 98% water and 2% acetonitrile are
9.5min；(2) 76% water and 24% acetonitrile are 1.5min；(3) 20% water and 80% acetonitrile 2min；(4) 20%
Water and 80% acetonitrile 5min；(5) 98% water and 2% acetonitrile 9min.
Preferably, the mass spectrographic condition is：70eV positive ion modes, carry out under room temperature condition.
Present invention also offers a kind of stomach cancer amino acid tags thing to combine the application in diagnosing gastric cancer, stomach cancer amino acidity scale
The combination of will thing includes following several or whole combinations：Taurine, glycine, glutamine, monoethanolamine, histidine, alanine,
Glutamic acid, hydroxylysine, proline and tyrosine.
Preferably, if the taurine, the glycine, the glutamine, the monoethanolamine, the histidine,
In the alanine, the glutamic acid, the proline and the tyrosine, at least 7 in the saliva sample of testee
The concentration of kind amino acid tags thing is more than 50 μM, then the testee suffers from stomach cancer.
Preferably, if the glycine, the alanine, the glutamic acid or the proline testee saliva
Concentration in sample is all higher than 100 μM, then the testee is in early gastric caacer.
Preferably, if the taurine, the glycine, the glutamine, the monoethanolamine, the histidine,
In the alanine, the glutamic acid, the proline and the tyrosine, at least 7 in the saliva sample of testee
The concentration of kind amino acid tags thing is between 50~100 μM, then the testee is in advanced gastric cancer.
Preferably, if the taurine, the glycine, the glutamine, the monoethanolamine, the histidine,
In the alanine, the glutamic acid, the hydroxylysine, the proline and the tyrosine, in the saliva of testee
The concentration of at least 8 kinds amino acid tags things is less than 50 μM in sample, then the testee does not have stomach cancer.
Principle and beneficial effect：Relation selection between metabolin small molecule and cancer in saliva is a kind of most appropriate
Metabolism small molecule as mark, be the key for utilizing saliva to carry out early gastric caacer examination and diagnosis.It is small relative to other
Metabolin, the amino acid in saliva are the materials of human body itself generation, are not easily susceptible to the influence of diet, while species is less and dense
Spend larger, chemical property stabilization；Most importantly, the amino acid in saliva can generate in stomach organization, and by true
Fixed intracellular and paracytic bang path is entered in saliva by stomach organization.Therefore, in early carcinoma of stomach and late gastric cancer
Diagnosis in, the amino acid in saliva is preferable biomarker, has very high accuracy.
Compared with prior art, the present invention has the advantages that：
1st, sample is collected and detection method is more convenient, simple, quick.Controllable on the collection capacity and acquisition time of saliva, side
Just, noninvasive close friend, in the absence of any ethics and medical-risk, detection process is no more than 2 hours.
2nd, the stomach cancer amino acid tags species filtered out in saliva are more, good with the relevance of stomach cancer, more by detecting
Individual stomach cancer amino acid tags thing can effectively improve the accuracy rate of diagnosing gastric cancer.
3rd, early carcinoma of stomach and advanced gastric carcinoma can be distinguished, is advantageous to determine clinical treatment, improves cure rate.
Brief description of the drawings
Amino acid tags thing is dense in the saliva sample that Fig. 1 is early carcinoma of stomach patient, late gastric cancer patient provides with healthy person
Spend relation schematic diagram.
Embodiment
The present invention is described in detail below in conjunction with the accompanying drawings.
The collection of saliva sample：
Saliva sample is collected by following three standards：(1) the clinical diagnosis result of stomach cancer, comprising has age, sex,
The information of Staging of Gastric Cancer；(2) other malignant neoplasms or hematologic disease must not be suffered from by providing the volunteer of saliva sample；(3) provide
The volunteer of saliva sample must not suffer from various metabolic system diseases, mainly including diabetes etc..In this research, 135 are shared
Volunteer, including 12 early carcinoma of stomach patients, 50 late gastric cancer patients and 73 Healthy Peoples provide saliva sample.All samples
Staging of Gastric Cancer information in this is by clinician according to clinical examination and Cancer Staging Handbook (AJCC Cancer Staging
Manual) judged.
Whole saliva sample gatherer process is as follows：Before sample collection, all volunteers must forbid in 1h
Any food is taken in keep oral cleaning.Before sampling starts, first gargled with pure water, start to collect after 5min, every volunteer
Need to gather 4mL saliva samples.Resulting saliva sample is in centrifuge to centrifuge 30min under the conditions of 12000rpm, 4 DEG C.
Then, careful taking-up 2mL supernatant liquors and centrifuge tube is transferred to, be stored in -70 DEG C of conditions, to carry out the analysis of next step inspection
Survey.
The screening of stomach cancer amino acid tags thing combination：
In order to filter out in saliva reliable amino acid as stomach cancer marker.The saliva sample of separate sources utilizes
TRAQTM kits (AB SCIEX, the U.S.), HPLC-MS technology (High performance liquid
Chromatography-mass spectrometry, HPLC-MS, AB SCIEX, the U.S.) analyzed.Wherein, efficient liquid phase
Chromatographic system mainly includes amino-acid analyzer, AB3200 QTRAP triple quadrupoles mass spectrographs and C18 liquid-phase chromatographic columns (4.6mm
Internal diameter × 150mm length, using C18 as packing material, granular size is 5.0 μm, AB SCIEX, the U.S.).During detection, initially
Equilibration time is 5min, and gradient activation is arbitrarily carried out by following condition：(1) 98% water and 2% acetonitrile are 9.5min；(2)
76% water and 24% acetonitrile are 1.5min；(3) 20% water and 80% acetonitrile 2min；(4) 20% water and 80%
Acetonitrile 5min；(5) 98% water and 2% acetonitrile 9min.Analysis run time is 25min, and Flow sample is 98% water and 2%
Acetonitrile, flow 0.8mL/min.Mass spectral analysis detection employs 70eV positive ion modes.All analysis results use software
Analyst 1.4.2 carry out automatic identification and quantified, and whole process is carried out at ambient temperature.Analysis result is shown in Table 1.
The amino acid detected in the saliva that table 1 comes from the collection of early carcinoma of stomach patient, late gastric cancer patient and Healthy People is dense
Degree
It can be seen that wherein have that 10 kinds of amino acid concentrations are considerably higher, this in 10 amino acid be respectively：Taurine, glycine, paddy
Glutamine, monoethanolamine, histidine, alanine, glutamic acid, hydroxylysine, proline and tyrosine, also, this amino acid in 10
Significant concentration difference is shown in the saliva of early carcinoma of stomach patient, late gastric cancer patient and Healthy People, than others 28
Kind amino acid is more suitable for as stomach cancer marker, therefore this 10 kinds of amino acid are selected in present embodiment as stomach cancer amino acid
Mark combines.
Generally, for the rate of metabolism in stomach organization apparently higher than normal structure, the synthesis for including amino acid is fast
Degree.Therefore, the amino acid tags thing concentration in patients with gastric cancer saliva is completely reasonable apparently higher than Healthy People.Meanwhile work as stomach
When cancer is in early stage, the accretion rate highest in stomach organization, therefore, the amino acid tags thing in early carcinoma of stomach patient's saliva
Concentration highest is also entirely sensible.In addition, the concentration of amino acid tags thing is also by cancer in amino acid transport system in saliva
The tissue influence different to this 10 kinds of amino acid tags thing affinity with health tissues.So this 10 kinds of organic volatiles are not only
Direct diagnosis of gastric cancer can be used for as the stomach cancer amino acid tags thing in saliva, can more be used to judging stomach cancer in early stage or
Progressive stage, aid in determining whether corresponding therapeutic scheme.
Stomach cancer amino acid tags thing combines the application in diagnosing gastric cancer：
Stomach cancer amino acid tags thing in obtained saliva is used for quick detection stomach cancer by the present embodiment, is carried out effect and is tested
Card：
Pass through detection, from 8 early carcinoma of stomach patients, saliva that 34 late gastric cancer patients and 43 healthy people collect
In liquid sample, when taurine, glycine, glutamine, monoethanolamine, histidine, alanine, glutamic acid, proline and tyrosine
When concentration in saliva is all higher than 50 μM, show that testee suffers from stomach cancer.When glycine, alanine, glutamic acid and dried meat ammonia
When concentration of the acid in saliva is all higher than 100 μM, show that the stomach cancer of testee is in early stage.When taurine, glycine, paddy ammonia
In acid amides, monoethanolamine, histidine, alanine, glutamic acid, proline and tyrosine, there are 7 kinds of amino acid tags things in saliva
Concentration shows that the stomach cancer of testee is in late period when between 50 to 100 μM, wherein having 3 kinds not in (this 7 kinds of Amino acid scores
It is not taurine, glycine, monoethanolamine, alanine, glutamic acid, proline and tyrosine).When taurine, glycine, glutamy
In amine, monoethanolamine, histidine, alanine, glutamic acid, hydroxylysine, proline and tyrosine, the concentration in saliva is respectively less than
At 50 μM, show subject without stomach cancer.
Detection process is same as above, and as a result finds there is 5 early carcinoma of stomach samples, 30 advanced gastric carcinoma samples and 32 healthy samples
Originally the conclusion of the above is met.
The technical concepts and features of above-mentioned embodiment only to illustrate the invention, its object is to allow be familiar with technique
People can understand present disclosure and implement according to this, and it is not intended to limit the scope of the present invention.It is all according to the present invention
The equivalent transformation or modification that Spirit Essence is done, should all be included within the scope of the present invention.
Claims (10)
1. a kind of stomach cancer amino acid tags thing combination, it is characterised in that including following several or whole combinations：It is taurine, sweet
Propylhomoserin, glutamine, monoethanolamine, histidine, alanine, glutamic acid, hydroxylysine, proline and tyrosine.
2. a kind of screening technique of stomach cancer amino acid tags thing combination, it is characterised in that utilize HPLC-MS
Instrument determines the metabolism amino acid in patients with gastric cancer and Healthy People saliva sample, and filters out the metabolism amino acid related to stomach cancer and make
Combined for stomach cancer amino acid tags thing；The stomach cancer amino acid tags thing combination includes following several or whole combinations：Ox sulphur
Acid, glycine, glutamine, monoethanolamine, histidine, alanine, glutamic acid, hydroxylysine, proline and tyrosine.
3. the screening technique of stomach cancer amino acid tags thing combination according to claim 2, it is characterised in that the efficient liquid
The condition of phase chromatogram is：From C18 chromatographic columns, Flow sample is 98% water and 2% acetonitrile, is percent by volume, is carried out
Gradient activates；Flow velocity is 0.8mL/min, and column temperature is room temperature, and the initial equilibration time is 5min, and analysis run time is 25min, room
Carried out under the conditions of temperature.
4. the screening technique of stomach cancer amino acid tags thing combination according to claim 3, it is characterised in that the gradient is lived
Change is arbitrarily carried out according to following condition：(1) 98% water and 2% acetonitrile are 9.5min；(2) 76% water and 24% acetonitrile
For 1.5min；(3) 20% water and 80% acetonitrile 2min；(4) 20% water and 80% acetonitrile 5min；(5) 98% water
With 2% acetonitrile 9min.
5. the screening technique of the stomach cancer amino acid tags thing combination according to any one of claim 2 to 4, its feature exist
In the mass spectrographic condition is：70eV positive ion modes, carry out under room temperature condition.
6. a kind of stomach cancer amino acid tags thing combines the application in diagnosing gastric cancer, it is characterised in that stomach cancer amino acid tags thing
Combination includes following several or whole combinations：Taurine, glycine, glutamine, monoethanolamine, histidine, alanine, paddy ammonia
Acid, hydroxylysine, proline and tyrosine.
7. stomach cancer amino acid tags thing according to claim 6 combines the application in diagnosing gastric cancer, it is characterised in that its
It is characterised by, if in the taurine, the glycine, the glutamine, the monoethanolamine, the histidine, described third
In propylhomoserin, the glutamic acid, the proline and the tyrosine, at least 7 kinds of amino in the saliva sample of testee
The concentration of sour mark is more than 50 μM, then the testee suffers from stomach cancer.
8. stomach cancer amino acid tags thing according to claim 6 combines the application in diagnosing gastric cancer, it is characterised in that if
The concentration of the glycine, the alanine, the glutamic acid and the proline in the saliva sample of testee is big
In 100 μM, then the testee is in early gastric caacer.
9. stomach cancer amino acid tags thing according to claim 6 combines the application in diagnosing gastric cancer, it is characterised in that if
In the taurine, the glycine, the glutamine, the monoethanolamine, the histidine, the alanine, the paddy
In propylhomoserin, the proline and the tyrosine, at least 7 kinds of amino acid tags things is dense in the saliva sample of testee
Degree is between 50~100 μM, then the testee is in advanced gastric cancer.
10. stomach cancer amino acid tags thing according to claim 6 combines the application in diagnosing gastric cancer, it is characterised in that
If in the taurine, glycine, the glutamine, the monoethanolamine, the histidine, the alanine, described
In glutamic acid, the hydroxylysine, the proline and the tyrosine, at least 8 kinds in the saliva sample of testee
The concentration of amino acid is less than 50uM, then the testee does not have stomach cancer.
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| CN110045035A (en) * | 2019-04-30 | 2019-07-23 | 上海交通大学 | Gastric cancer VOC marker in saliva and its preparing the application in stomach cancer diagnosis reagent |
| CN111540469A (en) * | 2020-05-29 | 2020-08-14 | 杭州广科安德生物科技有限公司 | Method for constructing mathematical model for in-vitro detection of gastric cancer and application thereof |
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| CN116386716A (en) * | 2023-06-06 | 2023-07-04 | 浙江省肿瘤医院 | Metabolomics and methods for gastric cancer diagnosis |
| CN116386716B (en) * | 2023-06-06 | 2024-03-15 | 浙江省肿瘤医院 | Metabolomics and methods for gastric cancer diagnosis |
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