CN109541209A - Esophageal squamous cell carcinoma microenvironment cell sign object molecular model and its application - Google Patents
Esophageal squamous cell carcinoma microenvironment cell sign object molecular model and its application Download PDFInfo
- Publication number
- CN109541209A CN109541209A CN201811123407.8A CN201811123407A CN109541209A CN 109541209 A CN109541209 A CN 109541209A CN 201811123407 A CN201811123407 A CN 201811123407A CN 109541209 A CN109541209 A CN 109541209A
- Authority
- CN
- China
- Prior art keywords
- esophageal squamous
- squamous cell
- cell carcinoma
- invasion
- sma
- 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.)
- Granted
Links
Classifications
-
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
-
- 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
- G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
- G01N33/531—Production of immunochemical test materials
- G01N33/532—Production of labelled immunochemicals
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Immunology (AREA)
- Engineering & Computer Science (AREA)
- Urology & Nephrology (AREA)
- Chemical & Material Sciences (AREA)
- Molecular Biology (AREA)
- Biomedical Technology (AREA)
- Hematology (AREA)
- Cell Biology (AREA)
- Medicinal Chemistry (AREA)
- Analytical Chemistry (AREA)
- Microbiology (AREA)
- Pathology (AREA)
- Food Science & Technology (AREA)
- Biotechnology (AREA)
- Physics & Mathematics (AREA)
- General Physics & Mathematics (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Hospice & Palliative Care (AREA)
- Oncology (AREA)
- Investigating Or Analysing Biological Materials (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
It mainly include a-SMA the present invention relates to a kind of esophageal squamous cell carcinoma microenvironment cell sign object molecular model and its application, molecular model+ (front of invasion)Fibroblast, CD163+ (interstitial)Macrophage, CD117+ (lamina propria)Mast cell, CD117+ (front of invasion)Mast cell；It can be used for predicting patients with esophageal squamous cell carcinoma prognosis and to the grouping of adjuvant treatment sensibility group.Kit mainly includes anti-CD117 protein antibodies, anti-CD163 protein antibodies and anti-a-SMA protein antibodies.The prognosis of 3 molecule binding of pathological of the present invention histologic characteristics associated prediction patient has higher prediction effect than single Indexs measure, also obviously distinguish esophageal squamous cell carcinoma simultaneously and assist in the treatment of sensitive patients, and the super quick type two step method of immunohistochemistry that is based on of the present invention be a kind of mature and reliable, can be in the widely used method of basic hospital.More increase with international TNM stage compared to prediction effect, it is simpler, easy, have more sensibility and specificity.
Description
Technical field
The present invention relates to the multiple protein antibodies of joint and Histopathological Features, and utilize the super quick type two of immunohistochemistry
Footwork removes the expression of detection patients with esophageal squamous cell carcinoma sample, and then predicts patient's prognosis and adjuvant treatment sensibility.
Background technique
Esophageal squamous cell carcinoma (Esophageal Squamous Cell Carcinoma, ESCC), abbreviation esophageal squamous cell carcinoma,
It is a kind of common upper-digestive-tract malignant tumors in China, the 4th is occupied in the mortality of malignant tumors of compatriots.The whole world
In annual more than 40 ten thousand new hair esophageal squamous cell carcinoma case, more than half occurs in China, and China is also esophageal squamous cell generally acknowledged in the world
The high-incidence state of cancer first.1970s implement malignant tumour generaI investigation in China, establish Cixian, hebei Province, Henan woods
The area such as county, Yangquan Shanxi, Sichuan Yanting, Guangdong Chaozhou-Shantou region and Jiangsu Huaian is the big district occurred frequently of Chinese esophageal squamous cell carcinoma six.
The prognosis of Accurate Prediction patients with esophageal squamous cell carcinoma and the sensibility of adjuvant treatment have clinical further decision
Significance.However, a kind of simple and effective method can be more accurately predicted not yet other than international TNM stage at present
The prognosis of patients with esophageal squamous cell carcinoma.However, the prognosis of TNM stage prediction patient is comparatively laborious, need accurately to evaluate the leaching of tumour
Moistening depth, the number of metastatic lymph node, tumour, whether there is or not DISTANT METASTASES INs etc., moreover the formulation of international TNM stage lacks a large amount of compatriots
The case-data of esophageal squamous cell carcinoma, therefore applicability is poor when evaluating the prognosis of China patient；In prediction Concurrent Chemoradiotherapy Sensitivity
Aspect is more the absence of available molecular marked compound.
A-SMA label cancer-related fibroblast, be a variety of malignant entity tumor microenvironments main interstitial at
Point, key effect is played in the process and treatment of tumour.Cancer-related fibroblast is due to its tissue-derived difference
Different cell factor and extracellular matrix can be secreted and play various different functions.
The macrophage of CD163 label is most common one kind inflammatory cell in tumor microenvironment, it is played in tumour
A variety of key effects, there is very high plasticity, or even affect the treatment of tumour.Have an article report, cancer-related at
Fibrocyte secretes the macrophage colony stimulating factor expression of IL-6 up-regulation monocyte, so that monocyte be made to be divided into
CD163+M2 type macrophage.
The mast cell of CD117 label is more controversial a kind of cell in tumor microenvironment, often in tumour
Play different or even opposite effect.
Summary of the invention
To solve the above problems, the present invention provides a kind of esophageal squamous cell carcinoma microenvironment cell sign object molecular models
And its application.
A kind of kit for predicting patients with esophageal squamous cell carcinoma prognosis, including anti-CD117 protein antibodies, anti-CD163 protein antibodies
With anti-a-SMA protein antibodies.
Present invention discover that in esophageal squamous cell carcinoma front of invasion, there are a subgroup cancer-related fibroblasts not to directly affect
Tumour cell EMT, and it is related to the remodeling of microenvironment and the activation of inflammatory cell, infiltration.CD163 in esophageal squamous cell carcinoma+Macrophage is thin
Born of the same parents' quantity is significant related to patient's poor prognosis, and is closely related with tumor size, tumor invasive depth.Esophageal squamous cell carcinoma invasion
Forward position fibroblast is positively correlated with macrophage quantity.The mast cell of esophageal squamous cell carcinoma lamina propria and the good prognosis of patient
Correlation, and the mast cell of front of invasion and the poor prognosis of patient are closely related.And front of invasion fibroblast and hypertrophy
Cell quantity is negatively correlated.
Although CD117 albumen, CD163 albumen and a-SMA albumen all have the ability of prediction patients with esophageal squamous cell carcinoma prognosis,
Tumor microenvironment is complicated whole as one, and cellular component is a dynamic change and interaction joint effect tumour occurs
Development and treatment.Therefore histological characteristic binding is combined to get up to predict the prognosis of patients with esophageal squamous cell carcinoma 3 kinds of relevant molecules
Then there is stronger judgement efficiency, the predictive ability of even higher than internationally recognized TNM stage, and esophageal squamous cell carcinoma can be assisted
Sensitive patient is treated to distinguish.
Wherein the detection reagent of kit is lowlenthal serum respectively, and 0.01M citrate repairs liquid, 3%H2O2,
Polymer reinforcing agent, Polymer polymer, DAB colour reagent and PBS solution.
The kit of above-mentioned prediction patients with esophageal squamous cell carcinoma prognosis is preparing the prognosis of associated prediction patients with esophageal squamous cell carcinoma and to auxiliary
Help the application of therapeutic sensitivity group group reagent box.
It wherein, the use of kit sample detected is the fixed paraffin embedded tissues with paraffin embedding of neutral formalin.
Wherein, used detection method is the super quick type two step method of immunohistochemistry, the specific steps are as follows: to neutral good fortune
The fixed paraffin embedded tissues with paraffin embedding of your Malin are sliced, and slice is dewaxed to aquation, antigen retrieval, inactivating endogenous mistake
It is anti-with anti-CD117 protein antibodies, anti-CD163 protein antibodies and anti-a-SMA albumen respectively after oxide enzyme and lowlenthal serum closing
Body 4 DEG C of overnight incubations in moisture preservation box；After being then incubated for respectively with Polymer reinforcing agent and Polymer polymer, DAB colour developing
Nucleus is redyed with haematoxylin；Neutral gum mounting is most used after dehydration of alcohol and dimethylbenzene are transparent afterwards, and every dyeing is cut
Piece scores.
Wherein, the anti-CD117 protein antibodies label esophageal squamous cell carcinoma lamina propria and the mast cell of front of invasion, it is described anti-
CD163 protein antibodies mark M2 type macrophage, the anti-a-SMA protein antibodies marked tumor correlation fibroblast, benefit
The mast cell number and mesenchyma stroma of tumors of lamina propria, front of invasion are calculated with the multispectral slice automatic analysis system of Vectra
(the dyeing of front of invasion a-SMA positive tumor correlation fibroblast expression value is differentiated under M2 type macrophage number and mirror
Intensity × positive area).The cut-off value of three kinds of albumen height expression determines by X-tile software, wherein the cut- of CD117
It is the cut-off value of 600, a-SMA is 5 points that off value, which is the cut-off value of 100, CD163,.
A kind of esophageal squamous cell carcinoma microenvironment cell sign object molecular model mainly includes a-SMA+ (front of invasion)At fiber
Cell, CD163+ (interstitial)Macrophage, CD117+ (lamina propria)Mast cell, CD117+ (front of invasion)Mast cell.
Wherein, the equation of molecular model is constructed are as follows: every sample after detecting respectively using 3 kinds of protein antibodies, and 3 kinds
The calculation formula of albumen Combined expression is Y=α × A+ β × B+ γ × C+ δ × D, and wherein α, β, γ, δ are coefficient, can be passed through
Cox recurrence obtains, therefore the formula is converted to Y=0.812 × A+1.788 × B+ (- 0.931) × C+0.967 × D；A, B, C, D points
It Wei not a-SMA in this sample+ (front of invasion)Fibroblast, CD163+ (interstitial)Macrophage, CD117+ (lamina propria)Mast cell and
CD117+ (front of invasion)The expression status of mast cell, 0 represents low expression, and 1 represents high expression；In the Y value for calculating every sample
Afterwards, patients with esophageal squamous cell carcinoma is divided into according to X-tile software by height-, in-, (cut-off value is respectively 1.80 Hes to low-risk group
2.76) it is high risk group that, i.e., Y value, which is greater than 2.76, and it is risk group that Y value, which is more than or equal to 1.80 and is less than or equal to 2.763, and Y value is small
It is low-risk group in 1.80.
The application of above-mentioned esophageal squamous cell carcinoma microenvironment cell sign object molecular model, can be used for predicting esophageal squamous cell carcinoma
Patient's prognosis and to adjuvant treatment sensibility group grouping.
It, only need to be 3 albumen of the patient when evaluating individual patients with esophageal squamous cell carcinoma prognosis and assisting in the treatment of the information of sensibility
ImmunohistochemistryResults Results corresponding to expression status substitute into formula in, so that it may obtain Y value, then with determining cut-off value
Compare.Obtain corresponding 1 year, 3 years, 5 years overall survivals and disease free survival, and corresponding surgery alone group and operation+auxiliary
The Survival for the treatment of group, while obtaining the prognosis suggestion of different therapeutic modalities.
Compared with prior art, 3 molecule binding of pathological of the present invention histologic characteristics associated prediction patient
Prognosis has higher prediction effect than single Indexs measure, while also obviously distinguishing esophageal squamous cell carcinoma adjuvant treatment sensibility and suffering from
Person, and the super quick type two step method of immunohistochemistry that is based on of the present invention be a kind of mature and reliable, can be extensive in basic hospital
The method used.More increase with international TNM stage compared to prediction effect, it is simpler, easy, with more sensibility and specifically
Property.
Detailed description of the invention
Fig. 1 is front of invasion a-SMA+Fibroblast is (total in the Kaplan-Meier survivorship curve of different expression crowds
Survival rate: P=0.002；Disease free survival: P=0.001).
Fig. 2 is interstitial CD163+Macrophage different expression crowds Kaplan-Meier survivorship curve (overall survival:
P<0.001；Disease free survival: P < 0.001).
Fig. 3 is lamina propria and front of invasion CD117+Mast cell is survived bent in the Kaplan-Meier of different expression crowds
Line；Lamina propria CD117+The high expression extension patients with esophageal squamous cell carcinoma prognosis of mast cell, and front of invasion CD117+Mast cell with
Related (the lamina propria: overall survival: P < 0.001, disease free survival: P=0.003 of patient's prognosis mala；Front of invasion: total existence
Rate: P=0.001, disease free survival: P=0.011).
Fig. 4 is front of invasion a-SMA+Fibroblast and interstitial CD163+Macrophage, CD117+The correlation of mast cell
Relationship；Front of invasion a-SMA+Fibroblast and interstitial CD163+Macrophage is positively correlated (R=0.239, P=0.001),
Front of invasion a-SMA+Fibroblast and lamina propria, front of invasion CD117+Negatively correlated (the lamina propria: R=- of mast cell
0.359, P < 0.001；Front of invasion: R=-0.207, P=0.003).
Fig. 5 is the Kaplan-Meier survivorship curve (overall survival: P < 0.001 that molecular model judges the crowd；No tumor is raw
Deposit rate: P < 0.001).
Fig. 6 is ROC curve；Area is apparently higher than TNM points under the ROC curve of molecular model overall survival and disease free survival
Phase.
Fig. 7 is the relationship of molecular model and treatment；Relative to low-risk group and risk group, high risk group operation+auxiliary
Treatment is obviously prolonged total existence of patients with esophageal squamous cell carcinoma and survives without tumor.
Specific embodiment
To make the object, technical solutions and advantages of the present invention clearer, the present embodiment has collected 207 2012-2014
The specimens from pri of the patients with esophageal squamous cell carcinoma in year achieves wax stone.The present invention will be describe below in further detail with reference to the accompanying drawings.
Steps are as follows for the super quick type two step method of immunohistochemistry:
Sample is after serial section, dimethylbenzene dewaxing, graded ethanol aquation, with super quick type two step method testing goal albumen
Expression.The specific method is as follows:
(1) slice repairs liquid antigen retrieval, 3%H through 0.01M citrate2O2Deactivating endogenous peroxydase, goat
After serum closing, it is incubated overnight respectively with 4 DEG C of a-SMA, CD163 and CD117 antibody；
(2) after slice is incubated for respectively with Polymer reinforcing agent and Polymer polymer, DAB colour developing；
(3) slice redyes core through haematoxylin, gradient alcohol dehydration, and dimethylbenzene is transparent and neutral gum mounting；
(4) slice is washed through PBS solution after completing above-mentioned every step.
All immunostained sections are independently scored through 2 Pathology Doctors 's using blind.A-SMA mainly assesses front of invasion
Positive fibroblasts, methods of marking are expression intensity × positive area.Staining power is divided into 4 grades: 0, feminine gender；1, weak dyeing；
2, moderate dyeing；3, it is strong to dye.Positive cell percentage is divided into 5 grades: 0,0~5%；1,6~25%；2,26~50%；3,51
~75%；4, >=76%.The gross score of each sample is by tumour cell staining power and positive tumor cell percentage two
The product divided show that range is 0~12.CD163+Macrophage and CD117+Mast Cells are by the multispectral slice of Vectra
Automatic analysis system calculates.According to the operating instruction of the multispectral slice automatic analysis system of the Vectra of standard, first at 40 times
Then automatically scanning histotomy under mirror acquires picture under 200 times of mirrors, pass through Vectra system automatic identification CD163+Macrophage is thin
Born of the same parents and CD117+Mast cell, it is then most to positive cell number before 20 pictures sum.It is in order to facilitate statistics, institute is ill
Example is divided into low expression and high 2 groups of expression, and 0 represents low expression group, and 1 represents high expression group, and cut-off value is according to X-tile software
It determines.A-SMA:0~5 point are low expression group, and 6~12 points are high expression group；CD163+Macrophage numbers: 0~600 is low table
Up to group, being greater than 600 is high expression group；CD117+Mast Cells: 0~100 is low expression group, and being greater than 100 is high expression group.
Front of invasion a-SMA+Fibroblast, interstitial CD163+Macrophage, lamina propria CD117+Mast cell and invasion
Forward position CD117+The equation that mast cell joins together to construct molecular model is as follows:
Y=α × A+ β × B+ γ × C+ δ × D, wherein α, β, γ, δ are coefficient, can be returned and be obtained by Cox, therefore the public affairs
Formula is converted to Y=0.812 × A+1.788 × B+ (- 0.931) × C+0.967 × D；A, B, C, D are respectively a- in this sample
SMA+(front of invasion) fibroblast, CD163+(interstitial) macrophage, CD117+(lamina propria) mast cell and CD117+
The expression status of (front of invasion) mast cell, 0 represents low expression, and 1 represents high expression；After the Y value for calculating every sample,
Patient with esophageal carcinoma is divided into height-according to X-tile software, in-, low-risk group (cut-off value is respectively 1.80 and 2.76), i.e.,
It is high risk group that Y value, which is greater than 2.76, and it is risk group that Y value, which is more than or equal to 1.80 and is less than or equal to 2.763, and Y value is less than 1.80
For low-risk group.Then single factor test multiplicity and ROC analysis are being carried out.
The statistical procedures of data:
Data are handled using 19.0 statistical software of SPSS.The relationship of the life span of the expression and patient of each interstitial component
Use Kaplan-Meier survival analysis；The independent hazard factor of predicted impact patient's prognosis uses Cox proportion risk regression mould
Type；The expression of each interstitial component and the relationship of clinicopathologic features are analyzed using Fisher's ExactTest；Every kind of component is pre-
The efficiency for surveying patients with esophageal squamous cell carcinoma prognosis uses ROC curve；P < 0.05 is considered having significance,statistical.
As the result is shown:
a-SMA+(front of invasion) fibroblast, CD163+(interstitial) macrophage, CD117+(lamina propria) is loose thin
Born of the same parents, CD117+The existent relationship of (front of invasion) mast cell and molecular model and patients with esophageal squamous cell carcinoma is shown in Fig. 1-3.Such as Fig. 1 institute
Show, front of invasion a-SMA+Fibroblastic high expression shortens total existence of patients with esophageal squamous cell carcinoma and survives without tumor.Fig. 2 existence
Analysis finds, CD163+(interstitial) macrophage is related to the poor prognosis of patients with esophageal squamous cell carcinoma.Fig. 3 is the results show that lamina propria
CD117+Mastocytosis is obviously prolonged total existence of patients with esophageal squamous cell carcinoma and survives without tumor, and front of invasion CD117+It is loose
The total of cell height expression group patients with esophageal squamous cell carcinoma survives and significantly reduces without tumor existence compared with low expression group.
As shown in figure 4, correlation analysis is found, front of invasion a-SMA+Fibroblastic expression and CD163+(interstitial)
The quantity of macrophage is proportional, with lamina propria CD117+Mast cell, front of invasion CD117+Mast cell is negatively correlated.
The discovery of Fig. 5 survival analysis, for molecular model high risk group relative to low, risk group is obviously shortened patients with esophageal squamous cell carcinoma
It is total existence and without tumor survive.
Fig. 6 ROC analysis finds, molecular model always survives to patients with esophageal squamous cell carcinoma and prediction efficiency without tumor existence is obviously high
In Ptnm-stage.
Fig. 7 existence chromatographic analysis discovery, molecular model high risk group is significant to adjuvant treatment to be benefited, the operation of high risk group
1 year overall survival of patient and disease free survival are respectively 12.5%, 0%；And the operation of high risk group+adjuvant treatment patient 1 year
Overall survival and disease free survival are respectively 66.7%, 37.5%.Molecular model high risk group patients with esophageal squamous cell carcinoma controls auxiliary
Treat obvious be benefited.
a-SMA+(front of invasion) fibroblast, CD163+(interstitial) macrophage, CD117+(lamina propria) is loose thin
Born of the same parents, CD117+The expression of (front of invasion) mast cell is all as shown in table 1 with the relationship of patients with esophageal squamous cell carcinoma clinicopathologic features:
1 result of table shows front of invasion a-SMA+Fibroblast and CD163+(interstitial) macrophage and tumor size, invasive depth phase
It closes；CD117+(lamina propria) mast cell and tumor size, invasive depth, lymphatic metastasis and clinical stages, are related.
The relationship of table 1 each marker and patients with esophageal squamous cell carcinoma clinicopathologic features
*Fisher's Exact Test；P value<0.05was considered significant.
a a-SMA(Invasive front of tumor),low,≤4scores；high,>4scores
b CD163(Stroma),low,≤600cells；high,>600cells
c CD117(Lamina propria),low,≤100cells；high,>100cells
d CD117(Invasive front of tumor),low,≤100cells；high,>100cells
The independent hazard factor of Cox risk forecast of regression model patient is as follows:
It from the analysis of table 2 as a result, it has been found that molecular model has stronger esophageal squamous cell carcinoma prognosis prediction ability, while being also independent
Prognostic factor.
The single argument and multi-variables analysis result of 2 patients with esophageal squamous cell carcinoma overall survival of table and disease free survival
Note: multiplicity, Cox proportional hazards regression models.The prognosis meaning of single factor analysis variable.
It, only need to be 3 albumen of the patient when information of the individual patients with esophageal squamous cell carcinoma prognosis of evaluation and auxiliary group of therapeutic sensitivity
ImmunohistochemistryResults Results corresponding to expression status substitute into formula in, so that it may obtain Y value, then with determining cut-off value
Compare, the Survival of corresponding each risk group is as shown in table 3:
The Survival of 3 207 each risk groups of patients with esophageal squamous cell carcinoma molecular model of table
Obtain corresponding 1 year, 3 years, 5 years overall survivals and disease free survival, and corresponding surgery alone group and operation+auxiliary
The Survival for the treatment of group is helped, while obtaining the prognosis suggestion of different therapeutic modalities.
The prediction efficiency of molecular model and each independent molecular marked compound, pTNM-stage compares
Find that molecular model is more preferably than single index, pTNM-stage prediction efficiency from table 4.
4 ROC of table analyzes result
Claims (10)
1. a kind of kit for predicting patients with esophageal squamous cell carcinoma prognosis, which is characterized in that including anti-CD117 protein antibodies, anti-CD163
Protein antibodies and anti-a-SMA protein antibodies.
2. predicting the kit of patients with esophageal squamous cell carcinoma prognosis according to claim 1, which is characterized in that further include blood of goats
Clearly, 0.01M citrate repairs liquid, 3%H2O2, Polymer reinforcing agent, Polymer polymer, DAB colour reagent and PBS are molten
Liquid.
3. predicting that the kit of patients with esophageal squamous cell carcinoma prognosis is preparing associated prediction patients with esophageal squamous cell carcinoma according to claim 1
Prognosis and to adjuvant treatment sensibility group group reagent box application.
4. applying according to claim 3, which is characterized in that sample detected is neutral formalin fixation and paraffin packet
The paraffin embedded tissues buried.
5. applying according to claim 3, which is characterized in that the detection method used is super quick two step of type of immunohistochemistry
Method, the specific steps are as follows: neutral formalin is fixed and the paraffin embedded tissues of paraffin embedding are sliced, slice is dewaxed to water
After change, antigen retrieval, deactivating endogenous peroxydase and lowlenthal serum closing, respectively with anti-CD117 protein antibodies, anti-
CD163 protein antibodies and anti-a-SMA protein antibodies 4 DEG C of overnight incubations in moisture preservation box；Then respectively with Polymer reinforcing agent and
After Polymer polymer is incubated for, DAB colour developing and haematoxylin redye nucleus；Most used after dehydration of alcohol and dimethylbenzene are transparent afterwards
Neutral gum mounting, and score every stained slice.
6. applying according to claim 5, which is characterized in that the anti-CD117 protein antibodies mark esophageal squamous cell carcinoma lamina propria
And the mast cell of front of invasion, the anti-CD163 protein antibodies mark M2 type macrophage, the anti-a-SMA protein antibodies
Marked tumor correlation fibroblast calculates lamina propria, front of invasion using the multispectral slice automatic analysis system of Vectra
Mast cell number and mesenchyma stroma of tumors M2 type macrophage number and mirror under differentiate front of invasion a-SMA positive tumor
Correlation fibroblast expression value；The cut-off value of CD117 albumen, CD163 albumen and the high low expression of a-SMA albumen is by X-
Tile software determines.
7. a kind of esophageal squamous cell carcinoma microenvironment cell sign object molecular model, which is characterized in that mainly include a-SMA+ (front of invasion)Fibroblast, CD163+ (interstitial)Macrophage, CD117+ (lamina propria)Mast cell, CD117+ (front of invasion)Mast cell.
8. esophageal squamous cell carcinoma microenvironment cell sign object molecular model according to claim 7, which is characterized in that building
The equation of molecular model are as follows: be Y=0.812 × A+1.788 × B+ (- 0.931) × C+0.967 × D；Wherein A, B, C, D points
It Wei not a-SMA+ (front of invasion)Fibroblast, CD163+ (interstitial)Macrophage, CD117+ (lamina propria)Mast cell and CD117+ (front of invasion)Fertilizer
The expression status of maxicell, 0 represents low expression, and 1 represents high expression；After calculating Y value, according to X-tile software esophageal squamous cell
Cancer patient is divided into height-, in-, it is high risk group that low-risk group, i.e. Y value, which are greater than 2.76, and Y value is more than or equal to 1.80 and is less than or equal to
2.763 be risk group, and Y value is low-risk group less than 1.80.
9. according to the application of the esophageal squamous cell carcinoma microenvironment cell sign object molecular model of claim 7 or 8, feature
It is, can be used for predicting patients with esophageal squamous cell carcinoma prognosis and to the grouping of adjuvant treatment sensibility group.
10. applying according to claim 9, which is characterized in that evaluate individual patients with esophageal squamous cell carcinoma prognosis and assist in the treatment of quick
When the information of perception, only need to expression status corresponding to the ImmunohistochemistryResults Results 3 albumen of patient substitute into formula, so that it may
To obtain Y value, then compared with determining cut-off value.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811123407.8A CN109541209B (en) | 2018-09-26 | 2018-09-26 | Molecular model of esophageal squamous cell carcinoma microenvironment cell marker and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811123407.8A CN109541209B (en) | 2018-09-26 | 2018-09-26 | Molecular model of esophageal squamous cell carcinoma microenvironment cell marker and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN109541209A true CN109541209A (en) | 2019-03-29 |
| CN109541209B CN109541209B (en) | 2022-07-08 |
Family
ID=65841190
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811123407.8A Active CN109541209B (en) | 2018-09-26 | 2018-09-26 | Molecular model of esophageal squamous cell carcinoma microenvironment cell marker and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN109541209B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110393800A (en) * | 2019-08-07 | 2019-11-01 | 石河子大学 | Medicine for enhancing chemotherapy sensitivity of esophageal squamous cell carcinoma |
| CN111269956A (en) * | 2020-02-25 | 2020-06-12 | 福建医科大学 | Application of reagent for detecting flora in preparation of reagent or kit for prognosis prediction marker of esophageal squamous cell carcinoma patient |
| CN111304325A (en) * | 2020-02-22 | 2020-06-19 | 深圳大学 | Oral cancer marker STOML1 gene expression and application thereof |
| CN111796101A (en) * | 2020-07-14 | 2020-10-20 | 汕头市中心医院 | Application of NOTCH3 protein expression amount detection agent and CYPA protein expression amount detection agent in prediction of cancer treatment curative effect |
| WO2023284789A1 (en) * | 2021-07-14 | 2023-01-19 | 郑州大学 | Molecular marker group of human esophageal squamous cell carcinoma and application of molecular marker group |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104407151A (en) * | 2014-11-19 | 2015-03-11 | 汕头大学医学院 | Kit integrating three proteins such as Kindlin-2, Myosin-9 and Annexin II for prognosis evaluation of patient suffering from esophageal squamous cell carcinoma |
| KR20160021970A (en) * | 2014-08-18 | 2016-02-29 | 사회복지법인 삼성생명공익재단 | Novel Biomarker for analysing prognosis of esophageal cancer |
| CN106153919A (en) * | 2016-06-16 | 2016-11-23 | 汕头大学医学院 | CD105, esVEGR2 and MYC tri-albumen associated prediction patients with esophageal squamous cell carcinoma prognosis kit |
| WO2017030929A1 (en) * | 2015-08-14 | 2017-02-23 | The Johns Hopkins University | Decoy nanoparticles to disrupt cancer cell-stormal cell networks |
-
2018
- 2018-09-26 CN CN201811123407.8A patent/CN109541209B/en active Active
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20160021970A (en) * | 2014-08-18 | 2016-02-29 | 사회복지법인 삼성생명공익재단 | Novel Biomarker for analysing prognosis of esophageal cancer |
| CN104407151A (en) * | 2014-11-19 | 2015-03-11 | 汕头大学医学院 | Kit integrating three proteins such as Kindlin-2, Myosin-9 and Annexin II for prognosis evaluation of patient suffering from esophageal squamous cell carcinoma |
| WO2017030929A1 (en) * | 2015-08-14 | 2017-02-23 | The Johns Hopkins University | Decoy nanoparticles to disrupt cancer cell-stormal cell networks |
| CN106153919A (en) * | 2016-06-16 | 2016-11-23 | 汕头大学医学院 | CD105, esVEGR2 and MYC tri-albumen associated prediction patients with esophageal squamous cell carcinoma prognosis kit |
Non-Patent Citations (2)
| Title |
|---|
| 周剑等: "食管鳞状细胞癌组织中M2型巨噬细胞浸润与患者预后及临床病理参数的相关性", 《新疆医科大学学报》 * |
| 樊慧杰: "C-kit和SCF在食管鳞癌中的表达及沉默c-kit基因表达对食管鳞癌细胞系KYSE70体外生物学行为的影响", 《中国博士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110393800A (en) * | 2019-08-07 | 2019-11-01 | 石河子大学 | Medicine for enhancing chemotherapy sensitivity of esophageal squamous cell carcinoma |
| CN111304325A (en) * | 2020-02-22 | 2020-06-19 | 深圳大学 | Oral cancer marker STOML1 gene expression and application thereof |
| CN111269956A (en) * | 2020-02-25 | 2020-06-12 | 福建医科大学 | Application of reagent for detecting flora in preparation of reagent or kit for prognosis prediction marker of esophageal squamous cell carcinoma patient |
| CN111269956B (en) * | 2020-02-25 | 2023-04-18 | 福建医科大学 | Application of reagent for detecting flora in preparation of reagent or kit for prognosis prediction marker of esophageal squamous cell carcinoma patient |
| CN111796101A (en) * | 2020-07-14 | 2020-10-20 | 汕头市中心医院 | Application of NOTCH3 protein expression amount detection agent and CYPA protein expression amount detection agent in prediction of cancer treatment curative effect |
| WO2023284789A1 (en) * | 2021-07-14 | 2023-01-19 | 郑州大学 | Molecular marker group of human esophageal squamous cell carcinoma and application of molecular marker group |
Also Published As
| Publication number | Publication date |
|---|---|
| CN109541209B (en) | 2022-07-08 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109541209A (en) | Esophageal squamous cell carcinoma microenvironment cell sign object molecular model and its application | |
| Minner et al. | Marked heterogeneity of ERG expression in large primary prostate cancers | |
| Glaessgen et al. | Heat shock proteins 27, 60 and 70 as prognostic markers of prostate cancer | |
| Laprie et al. | MIB‐1 immunoreactivity correlates with biologic behaviour in canine cutaneous melanoma | |
| CN106153922B (en) | A kind of colon cancer prognosis prediction mark and its detection method | |
| CN108107216B (en) | Application of combined marker in preparation of pancreatic cancer prognosis kit and determination system and method thereof | |
| Burdelski et al. | p16 upregulation is linked to poor prognosis in ERG negative prostate cancer | |
| Connolly et al. | Role of the Surgical Pathologist in the Diagnosis and Management of the Cancer Patient | |
| Pearl et al. | Stratification of patients by melanoma cell adhesion molecule (MCAM) expression on the basis of risk: implications for sentinel lymph node biopsy | |
| Mahajan et al. | AWARDS | |
| Di Napoli et al. | Tissue biomarkers in renal cell carcinoma: issues and solutions | |
| Brajkovic et al. | Microfluidics for rapid cytokeratin immunohistochemical staining in frozen sections | |
| Guo et al. | Contribution of bladder cancer pathology assessment in planning clinical trials | |
| Bol et al. | Proliferation markers and DNA content analysis in urinary bladder TaT1 urothelial cell carcinomas: identification of subgroups with low and high stage progression risks | |
| CN104034902A (en) | Kit for utilizing combination of four kinds of protein for predicting prognosis of esophagus cancer patient | |
| CN106153919B (en) | Tri- albumen associated prediction patients with esophageal squamous cell carcinoma prognosis kit of CD105, esVEGR2 and MYC | |
| Boruah et al. | Correlation of microvessel parameters in invasive ductal carcinoma of the breast and fibroadenomas: a morphometric study | |
| Shiba et al. | Prognostic implication of Ki-67 immunostaining in treating subclinical pleural cancer found at thoracotomy in lung cancer patients | |
| Pertia et al. | Loss of p27 (Kip1) CDKI is a predictor of poor recurrence-free and cancer-specific survival in patients with renal cancer | |
| Nambirajan et al. | AWARDS | |
| El-Mansi et al. | Validation of tissue microarray technology using cervical adenocarcinoma and its precursors as a model system | |
| Billis et al. | Prostate total tumor extent versus index tumor extent—which is predictive of biochemical recurrence following radical prostatectomy? | |
| Duderstadt et al. | A method to pre-screen rat mammary gland whole-mounts prior to RNAscope | |
| CN111665358A (en) | Application of NALCN protein in prognosis prediction of esophageal squamous cell carcinoma | |
| Tatlidil et al. | Detection of tyrosinase mRNA in the sentinel lymph nodes of melanoma patients is not a predictor of short-term disease recurrence |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |