CN107831308A - Transcript profile and the ImmunohistochemistryMethods Methods of protein science in a kind of liver cancer biological process - Google Patents
Transcript profile and the ImmunohistochemistryMethods Methods of protein science in a kind of liver cancer biological process Download PDFInfo
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- CN107831308A CN107831308A CN201711169079.0A CN201711169079A CN107831308A CN 107831308 A CN107831308 A CN 107831308A CN 201711169079 A CN201711169079 A CN 201711169079A CN 107831308 A CN107831308 A CN 107831308A
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- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
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- G01N33/57438—Specifically defined cancers of liver, pancreas or kidney
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Abstract
The invention discloses transcript profile and the ImmunohistochemistryMethods Methods of protein science in a kind of liver cancer biological process, wash 1. histotomy dewaxes, 2. aquation is added dropwise, is stored at room temperature 3. PBS, heat 0.01 sodium citrate buffer, it is put into tissue to carry out boiling hot reparation 4. PBS is washed and Normal Goat Serum closing is 5. added dropwise 6. is got rid of surplus liquid 7. PBS washes 8. PBS or running water and rinses 10 minutes 9. haematoxylin redyeings, 10. dehydration, transparent, mounting, microscopy are rinsed in hydrochloride alcohol differentiation.The present invention carries out the expression checking of positioning and quantitative using clinical sample to it, finds its evidence with clinical correlation, evaluates clinical value, and new clue is provided for onset of liver cancer and liver cancer Mechanism Study.The liver cancer key molecule of problem screening will establish Research foundation for the exploration liver cancer marker related to early detection, classification, evaluation prognosis, and more efficient, the accurate liver cancer treatment target position of selection.
Description
Technical field
The present invention relates to genetic transcription group and proteomics field, transcribed in especially a kind of liver cancer biological process
The ImmunohistochemistryMethods Methods of group and protein science.
Background technology
In biology and medical research, it is important that a field be to biosystem and the structure of life process, work(
The observation that can and regulate and control.But between the past centuries, biologist focuses on individual gene or protein in biology department always
Expression change and function in system, and the change of life system can not be studied from overall situation, the overall angle.With medical science
It is progressive, it has been found that the generations of many diseases, particularly cancer is often multifactor, polygenes, multipath synergy cause
's.This just need one can comprehensively, dynamic, the technology and means of systematic research life system, then " group is learned " is general
Thought is arisen at the historic moment[3].But with the completion of the Human Genome Project, it has been found that in only can not be complete from the angle of genomics
The shearing that occurs in total correctness predicted gene transcription, splicing and in translation the starting of open reading frame codon,
Various modification situations after final position and translation.
In gene expression research, extensive genetic analysis can be relevant to a physiological status either cell phenotype
Gene progress system monitoring, high throughput analysis can be utilized in data output and obtain the quick both sides advantage of data, to disease
Function candidate gene during disease is identified.The maturation of microarray technology, researcher is sequenced by transcript profile and study,
Find marker gene interested.As oncogene expression is to the tissue in various sources and the correlation point of patient's survival outcome
Analysis example is the same, and the gene expression analysis research carried out by microarray technology will continue to play the part of in biomarker discovery procedure
Important function.
Although the analysis ability of microarray is very powerful, transcription group research platform only includes the change of those Adaptable growth conditions
The transcript of cell.In most cells and intercellular Biochemical processes all can by protein-protein or other
The influence of protein-substrate interaction.The horizontal gene expression analysis of protein group provides a quickly controllable life
The process of thing synthesis, wherein most are regulated and controled by transcription group platform.Meanwhile transcript profile passes through the protein of expression in itself
Other changes either under cellular biochemical state, carry out feedback control.
In other words, gene expression is not exclusively from transcript profile to the one-way flow of protein group, but both is mutual
Connection.Understanding to this function controlling is generally limited to special signal pathway, or metabolic pathway.It is to be understood that turn
Effect of Mutual Regulation between record group and protein group to RNA and protein expression, it is necessary to carry out Integral synchronous monitoring.
The progress of transcription group, proteomics and bioinformatics investigative technique opens for research complex biological system
Brand-new approach, the reorganizing research that three is connected together can reveal that the hereditary information carried when disease occurs from gene turns
It is changed into that the exception during the entire process of phenotype can be distinguished, its magnanimity information gathered is covered in disease incidence and disease mechanisms
Key function node, can be used to identify tumor-related gene and its protein of expression so that thousands of genes and egg
The analysis of white matter is possibly realized, for explore early detection, classification, evaluate prognosis tumor markers, and selection it is more efficient,
Accurate oncotherapy target position provides reliable guarantee.
Ion proton sequenators of new generation use the technology of semiconductor chips, and sequencing speed is fast, and has high extension
Property, by proprietary large-scale parallel semiconductor inductor, ion stream caused by DNA replication dna is realized and directly and in real time examined
Survey.When reagent is entered in chip by integrated fluid passage, the reacting hole being clouded on chip immediately becomes up to a million individual micro-
Reaction system.The technical combinations of this unique fluid system, the Machine Design of microbody system and semiconductor, enable researcher to exist
The pinpoint accuracy sequence more than from 10Mb to 1Gb is obtained in 2 hours.In addition, Ion Proton sequenators and Ion Reporter
Analysis software can complete the analysis of individual gene group in an independent server, break current data parsing bottleneck, greatly
Research cost is reduced greatly, improves the speed and accuracy of detection, in scientific research and clinically there is good application；To current
Untill, in the confluence analysis article delivered, most of LC-MS analyses are used in combination with cold labeling, especially
It is iTRAQ reagents.Even with technology it is different, the confluence analysis published so far all indicate transcription group and
The importance of protein science.Transcription group or protein science generally only consider regulating system and the net effect of decomposition equilibrium state
Should, in fact, the inconsistency occurred is to synthesize a kind of reflection with two kinds of replacement process of degraded, researcher was to changing
Mechanism in journey is interested；In addition, transcription group and proteomic assays want successful integration, it is necessary to efficiently and accurately phase
Mutually reference.Researcher needs flexibly to define the genome of oneself, it is also possible to needing to select to be directed to using predefined
The target figure of protein, when new genome, transcript profile and protein groups sequence occur, researcher needs timely register update,
And the information of deletion error.The development of bioinformatics technique is so that genetic transcription, expression during oncobiology are whole
Exception during individual is disclosed, and clue is provided for tumour Mechanism Study.
This research is intended, using the sequencing of Ion Proton transcript profiles and LC-MALDI Discrepancy proteome analysis platforms, carrying out liver cancer
Transcript profile and proteomic assays in biological process.By building Rats With Hepatoma model, in relatively more normal and liver cancer tissue
Genetic transcription and protein expression difference, all occur to transcript profile in liver cancer and protein groups abnormal molecule carry out gene optimization,
Alternative splicing analysis, new gene or the screening of new transcript, expression analysis, Differential expression analysis, differential expression cluster analysis and
The processing of the bioinformatic analysis such as functional annotation, screens liver cancer key function node and tumor cells, and carry out clinic to it and test
Card and clinical value are assessed.This research will provide new clue for onset of liver cancer and liver cancer Mechanism Study.
Transcript profile and the SABC of protein science are compositions indispensable in the research in liver cancer biological process
Part, the technical scheme of transcript profile and the ImmunohistochemistryMethods Methods of protein science in an a kind of of the invention liver cancer biological process, warp
Retrieval domestic pharmaceutical industry industry has no identical.
The content of the invention
It is an object of the invention to provide transcript profile and the ImmunohistochemistryMethods Methods of protein science in a kind of liver cancer biological process.
Transcript profile and the ImmunohistochemistryMethods Methods of protein science in this liver cancer biological process,
Comprise the steps of：
1. histotomy dewaxing, after aquation PBS wash 2~3 times it is each 5 minutes；
2. 3% H2O2 (80% methanol) is added dropwise on TMA, it is stored at room temperature 10 minutes；
3. PBS wash 2~3 times it is each 5 minutes；Electric furnace or water-bath heat 0.01 sodium citrate buffer（pH 6.0）To 95 DEG C of left sides
The right side, it is put into the boiling hot reparation of tissue piece heating 10-15 minutes progress；
4. PBS wash 2~3 times it is each 5 minutes；
5. Normal Goat Serum confining liquid, room temperature 20 minutes is added dropwise；
6. get rid of surplus liquid；1 anti-50 μ l are added dropwise, be stored at room temperature 1 hour either 4 DEG C overnight or 37 DEG C 1 hour（Wherein, 4 DEG C
Need to be 45 minutes in 37 DEG C of rewarmings after overnight）；
7. PBS wash 3 times it is each 5 minutes；Secondary antibody is added dropwise（0.05% tween-20 can be added）40~50 μ l, are stored at room temperature, or 37 DEG C
1 hour;PBS wash 3 times it is each 5 minutes;DAB develops the color 5~10 minutes, grasps dye levels under the microscope；
8. PBS or running water rinse 10 minutes；
9. haematoxylin redyeing 2 minutes, hydrochloride alcohol differentiation；
10. running water rinses 10~15 minutes；Dehydration, transparent, mounting, microscopy.
Invention beneficial effect：
The present invention carries out the expression checking of positioning and quantitative using clinical sample to it, finds its evidence with clinical correlation, comments
Valency clinical value, new clue is provided for onset of liver cancer and liver cancer Mechanism Study.Problem screening liver cancer key molecule will be
Explore the liver cancer marker related to early detection, classification, evaluation prognosis, and more efficient, the accurate liver cancer treatment of selection
Target position establishes Research foundation.
Embodiment
Embodiment：
Transcript profile and the ImmunohistochemistryMethods Methods of protein science in this liver cancer biological process,
Comprise the steps of：
1. histotomy dewaxing, after aquation PBS wash 2~3 times it is each 5 minutes；
2. 3% H2O2 (80% methanol) is added dropwise on TMA, it is stored at room temperature 10 minutes；
3. PBS wash 2~3 times it is each 5 minutes；Electric furnace or water-bath heat 0.01 sodium citrate buffer（pH 6.0）To 95 DEG C of left sides
The right side, it is put into the boiling hot reparation of tissue piece heating 10-15 minutes progress；
4. PBS wash 2~3 times it is each 5 minutes；
5. Normal Goat Serum confining liquid, room temperature 20 minutes is added dropwise；
6. get rid of surplus liquid；1 anti-50 μ l are added dropwise, be stored at room temperature 1 hour either 4 DEG C overnight or 37 DEG C 1 hour（Wherein, 4 DEG C
Need to be 45 minutes in 37 DEG C of rewarmings after overnight）；
7. PBS wash 3 times it is each 5 minutes；Secondary antibody is added dropwise（0.05% tween-20 can be added）40~50 μ l, are stored at room temperature, or 37 DEG C
1 hour;PBS wash 3 times it is each 5 minutes;DAB develops the color 5~10 minutes, grasps dye levels under the microscope；
8. PBS or running water rinse 10 minutes；
9. haematoxylin redyeing 2 minutes, hydrochloride alcohol differentiation；
10. running water rinses 10~15 minutes；Dehydration, transparent, mounting, microscopy.
The foregoing is only a preferred embodiment of the present invention, but protection scope of the present invention be not limited thereto,
Any one skilled in the art the invention discloses technical scope in, technique according to the invention scheme and its
Inventive concept is subject to equivalent substitution or change, should all be included within the scope of the present invention.
Claims (1)
1. transcript profile and the ImmunohistochemistryMethods Methods of protein science in a kind of liver cancer biological process, it is characterised in that：By following step
Rapid composition：
1. histotomy dewaxing, after aquation PBS wash 2~3 times it is each 5 minutes；
2. 3% H2O2 (80% methanol) is added dropwise on TMA, it is stored at room temperature 10 minutes；
3. PBS wash 2~3 times it is each 5 minutes；Electric furnace or water-bath heat 0.01 sodium citrate buffer（pH 6.0）To 95 DEG C of left sides
The right side, it is put into the boiling hot reparation of tissue piece heating 10-15 minutes progress；
4. PBS wash 2~3 times it is each 5 minutes；
5. Normal Goat Serum confining liquid, room temperature 20 minutes is added dropwise；
6. get rid of surplus liquid；1 anti-50 μ l are added dropwise, be stored at room temperature 1 hour either 4 DEG C overnight or 37 DEG C 1 hour（Wherein, 4 DEG C
Need to be 45 minutes in 37 DEG C of rewarmings after overnight）；
7. PBS wash 3 times it is each 5 minutes；Secondary antibody is added dropwise（0.05% tween-20 can be added）40~50 μ l, are stored at room temperature, or 37 DEG C
1 hour;PBS wash 3 times it is each 5 minutes;DAB develops the color 5~10 minutes, grasps dye levels under the microscope；
8. PBS or running water rinse 10 minutes；
9. haematoxylin redyeing 2 minutes, hydrochloride alcohol differentiation；
10. running water rinses 10~15 minutes；Dehydration, transparent, mounting, microscopy.
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|---|---|---|---|
| CN201711169079.0A CN107831308A (en) | 2017-11-22 | 2017-11-22 | Transcript profile and the ImmunohistochemistryMethods Methods of protein science in a kind of liver cancer biological process |
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| CN201711169079.0A CN107831308A (en) | 2017-11-22 | 2017-11-22 | Transcript profile and the ImmunohistochemistryMethods Methods of protein science in a kind of liver cancer biological process |
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| CN107831308A true CN107831308A (en) | 2018-03-23 |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110331225A (en) * | 2019-07-11 | 2019-10-15 | 中国长江三峡集团有限公司 | One kind forming analysis method based on transcription group binding protein group TMT research lotus flower spot |
| CN111812328A (en) * | 2020-07-20 | 2020-10-23 | 邓鑫 | Kit for early detection and diagnosis of liver cancer metastasis |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104017868A (en) * | 2014-05-27 | 2014-09-03 | 江苏新昇生物技术有限公司 | Application of SETD4 to preparation of pancreatic cancer diagnosis and / or prognosis kit and application of SETD4 blocker to preparation of medicament for treating pancreas cancer |
-
2017
- 2017-11-22 CN CN201711169079.0A patent/CN107831308A/en active Pending
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104017868A (en) * | 2014-05-27 | 2014-09-03 | 江苏新昇生物技术有限公司 | Application of SETD4 to preparation of pancreatic cancer diagnosis and / or prognosis kit and application of SETD4 blocker to preparation of medicament for treating pancreas cancer |
Non-Patent Citations (2)
| Title |
|---|
| 郭素平: "Galectin-9和MMP-2蛋白在食管鳞癌中的表达及与预后的相关性", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
| 陈喜林: "原发性肝细胞癌肿瘤标志物的蛋白质组学研究", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110331225A (en) * | 2019-07-11 | 2019-10-15 | 中国长江三峡集团有限公司 | One kind forming analysis method based on transcription group binding protein group TMT research lotus flower spot |
| CN111812328A (en) * | 2020-07-20 | 2020-10-23 | 邓鑫 | Kit for early detection and diagnosis of liver cancer metastasis |
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