CN107367619A - Application, Prognosis in Breast Cancer assessment kit and method of the TCP1 albumen in Mammary cancer prognosis evaluation reagent kit is prepared - Google Patents
Application, Prognosis in Breast Cancer assessment kit and method of the TCP1 albumen in Mammary cancer prognosis evaluation reagent kit is prepared Download PDFInfo
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- CN107367619A CN107367619A CN201710759222.5A CN201710759222A CN107367619A CN 107367619 A CN107367619 A CN 107367619A CN 201710759222 A CN201710759222 A CN 201710759222A CN 107367619 A CN107367619 A CN 107367619A
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/68—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids
Abstract
The invention provides application, Prognosis in Breast Cancer assessment kit and method of the TCP1 albumen in Mammary cancer prognosis evaluation reagent kit is prepared.Relative expression quantity of the present invention using ImmunohistochemistryMethods Methods detection TCP1 albumen in breast cancer tissue, judges that the risk of breast cancer relapse transfer occurs in patient with breast cancer with this.The beneficial effects are mainly as follows：The invention provides application of the TCP1 albumen in Mammary cancer prognosis evaluation reagent kit is prepared, prompt the albumen to be used to prepare the protein molecular marker for judging patient with breast cancer's prognosis, also there is important directive significance for the postoperative monitoring of breast cancer patients and sequential therapy.
Description
Technical field
The invention belongs to biological technical field, and in particular to TCP1 albumen is preparing Mammary cancer prognosis evaluation reagent kit
In application, Prognosis in Breast Cancer assess kit and method.
Background technology
Breast cancer is one of most common malignant tumour of women, and according to statistics, the incidence of disease accounts for the various malignant tumours of whole body
7-10%, uterine cancer is only second in female diseases, the morbidity of breast cancer is often relevant with heredity, and patient is common in 40-60 year
Women between, women's incidence of disease before and after climacteric is higher, and only about 1-2% mammary gland patient is male.Breast cancer is usual
The malignant tumour in breast galandular epithelium tissue occurs, is a kind of to have a strong impact on the most common of the physically and mentally healthy even threat to life of women
One of malignant tumour.
The cause of disease of breast cancer is not yet fully apparent from, and research finds that the morbidity of breast cancer has certain regularity, has breast
The women of gland cancer high risk factor easily suffers from breast cancer.So-called high risk factor refer to it is relevant with pathogenesis of breast carcinoma it is various it is dangerous because
Element, and the hazards that most of patient with breast cancers have are known as the high risk factor of breast cancer.The early detection of breast cancer,
Early diagnosis, it is the key for improving curative effect.
Operative treatment is still one of essential therapeutic arsenals of breast cancer, and art formula has a variety of, selects it still weary unified opinion,
Total development trend is to reduce surgical disruption as far as possible, allows lower Remainded mamma of being done the best to early-stage breast cancer patient in appointed condition
Profile, no matter from which kind of art formula, it is necessary to strictly grasp based on effecting a radical cure, the principle supplemented by reservation function and profile.
The occurrence and development and prognosis of breast cancer and the activation of oncogene and tumor suppressor gene functionally inactive close association, be it is more because
Element induction, the complicated pathologic process that polygenes participates in.In recent years, the molecular mechanism of breast cancer early stage relapse and metastasis is the field
Heat subject, the molecular mechanism is deeply illustrated, and magnetic target therapy measure is imported in its specific link, be expected to very significantly
Improve Mammary cancer overall treatment effect.Therefore, it is related to seek the postoperative early stage recurrence of effective breast cancer breast excision
Early warning molecular marker, its relation with breast cancer early stage relapse and metastasis is illustrated, this is to improving Mammary cancer therapeutic effect, assessing
Breast cancer early stage risk of recurrence, judging prognosis and individualized treatment have vital meaning.
Protein folding is one of basic problem of life science.The folding of protein in the cell needs molecular chaperones
Assist.Therefore protein folding procedure is not completely spontaneous, and is a need energy process.Molecular chaperones can not only prevent
Protein forms aggregation or inactive structure in folding process, improves correct folding ratio, and can also affect on and fold road
Footpath.Molecular chaperones is mainly made up of three highly conserved protein familieses, and the member of these three families is distributed widely in protokaryon
In eukaryotic.TCP1 (t-complex polypeptide-1) is the chaperone in eukaryotic cytosol.TCP1
Actin and tubulin is assisted to unfold.
Breast cancer is as one of maximum tumour is threatened human health, and the molecular mechanism of its generation is still unclear so far,
Treatment to breast cancer also lacks specific molecular target, and TCP1 there is no at present as highly important tumour oncogene
Document report TCP1 albumen is related to judging Prognosis in Breast Cancer or Metastasis in Breast Cancer.And present invention research is found in breast cancer
TCP1 expresses has significant relation with patient's Postoperative determination, implies that TCP1 can be as effective early warning of the Postoperative determination of breast cancer
Albumen.
The content of the invention
Present invention aims at application of the offer TCP1 albumen in Mammary cancer prognosis evaluation reagent kit is prepared, and carry
For the kit and method for Mammary cancer prognosis evaluation.
The present invention has found by in-depth study extensively, TCP1 albumen is detected in breast using ImmunohistochemistryMethods Methods first
Relative expression quantity in adenocarcinoma tissue, it can interpolate that the risk of breast cancer relapse transfer occurs in patient with breast cancer.Based on TCP1 eggs
The correlation that white expression quantity shifts with breast cancer relapse, carrying out detection to its expression quantity as prognostic markers thing using TCP1 albumen can
For instructing the Index for diagnosis of breast cancer.
Therefore, the application the invention provides TCP1 albumen in Mammary cancer prognosis evaluation reagent kit is prepared, should answer
By the use of be using TCP1 albumen as molecular labeling, it is real with reference to SABC using TCP1 monoclonal antibodies or TCP1 polyclonal antibodies
Test reagent, relative expression quantity of the detection TCP1 albumen in breast cancer tissue.
Present invention also offers the kit for being used for Mammary cancer prognosis evaluation in the application, the kit includes：
People source TCP1 monoclonal antibodies or TCP1 polyclonal antibodies, immunohistochemical experiment reagent.The immunohistochemical experiment reagent is this
Common agents in the immunohistochemical experiment of field.
The present invention has found that TCP1 is expressed in recurrent and metastatic breast cancer tissue is higher than non-recurrent and metastatic breast cancer by research
Tissue, it can speculate that TCP1 plays a significant role in breast cancer recurrence transfer.Consult domestic and foreign literature, TCP1 and mammary gland
The generation of cancer and the correlative study of relapse and metastasis are few, in our current research, up-regulated expressions of the TCP1 in breast cancer tissue, and
Then expression is lowered by cancer and in normal liver tissue, and compared with non-relapse and metastasis group, TCP1 is expressed in relapse and metastasis group on also
Adjust, show that TCP1 participates in breast cancer occurrence and development process possibly as promotive factor.Pass through Kaplan-Meier survivorship curves point
Analysis, TCP1 expression degree is relevant with the prognosis of patient with breast cancer, patient's prognosis mala (P of the high expression of TCP1<0.05).
In summary, TCP1 high expression in breast cancer tissue, TCP1 high expression and patient with breast cancer's postoperative recurrence turn
Move relevant.TCP1 can be as an important candidate molecular marker thing of Prognosis in Breast Cancer.
Preferably, the people source TCP1 polyclonal antibodies are the TCP1 protein immunization rabbits shown in SEQ ID NO.1 as sequence
Son obtains, and can voluntarily prepare, can also use commodity purchased in market.
Immunohistochemical experiment reagent of the present invention includes：Dimethylbenzene, ethanol, 3%H2O2(aqueous solution), 3%BSA are closed
Liquid (being prepared with PBS), DAB colour reagents, haematoxylin, horseradish peroxidase (be used for mark secondary antibody), PBS (pH7.4),
0.01M EDTA repair liquid.
The method that Mammary cancer prognosis evaluation is carried out using kit of the present invention is as follows：
(1) patient with breast cancer's Pathologic specimen is taken, using people source TCP1 monoclonal antibodies or people source TCP1 polyclonal antibodies, with
And immunohistochemical experiment reagent, carry out immunohistochemical staining；
Wherein, Pathologic specimen comes from the pathologic sampling in patient with breast cancer's biopsy or art, postoperative.SABC side
Method utilizes SP decoration methods, comprises the following steps that：
(a) breast cancer tissue's paraffin section is prepared, 60 DEG C of baking boxs are stayed overnight.
(b) section dewaxing.Soak successively：Dimethylbenzene I：10min；Dimethylbenzene II：10min；Dimethylbenzene III：10min.
(c) section aquation.Soak successively：Absolute ethyl alcohol：3min；90% (v/v) ethanol：3min；80% ethanol：3min；
75% ethanol：3min.
(d) PBS 3 times, each 5min.
(e) EDTA antigens Pressure method：Section is put into 0.01M EDTA and repairs liquid immersion, boiling water bath 5min, is cooled to room
Temperature.PBS 3 times, each 5min.
(f) 300 μ L 3% (w/w) aqueous hydrogen peroxide solution, 37 DEG C of 10min are added.PBS 3 times, each 5min.
(g) 300 μ L 3% (w/w) BSA confining liquids (PBS preparations), 37 DEG C of 1h are added.PBS 3 times, each 5min.
(h) primary antibody is added：TCP1 antibody concentrations：1：500,4 DEG C of refrigerators take out after placing 16h, room temperature rewarming 15min, so
PBS is washed 4 times afterwards, each 5min.
(i) secondary antibody is added dropwise, described secondary antibody is that horseradish peroxidase-labeled goat anti-rabbit igg (steps novel agent purchased from Foochow
Company, instant, without dilution), 37 DEG C of 45min.PBS is washed 4 times, each 5min.
(j) PBS is washed 3 times, each 5min.Develop the color DAB (DAB colour reagent boxes, work being given birth to purchased from Shanghai) 2-10min, under mirror
Observation；Distilled water, which is washed, only to develop the color, and haematoxylin redyes 10s, is rinsed and soaked with running water.
(k) it is dehydrated.Soak successively：75% ethanol：2min；80% ethanol：2min；90% ethanol：2min；Absolute ethyl alcohol：
2min。
(L) electricity consumption blowing drying, neutral gum, cover glass covering are added.
(2) 3 visual field shootings of breast cancer tissue and cancer beside organism are randomly selected using microscope and imaging device, utilized
Aperio Image Scope softwares are scanned to the photograph of tissue samples, and the Algorithms of the software is used after scanning
(Positive Pixel Count V9) program carries out positive strength calculating to each sample, and it is as follows to calculate data：
(3) the immunohistochemistry score calculation of each tissue samples is Positivity × Log10 [255/Iavg], its
Middle Positivity=NPositive/NTotal, i.e. positive rate, computational methods are positive pixels quantity/colour developing total quantity；
Iavg=(Iwp+Ip+Isp)/(Nwp+Np+Nsp), i.e., positive mean intensity, computational methods are positive mean intensity=(weak sun
Property pixel overall strength+positive pixels overall strength+strong positive pixel overall strength)/(weakly positive pixel number+positive pixels quantity+strong
Positive pixels quantity), the as SABC scoring of the tissue, for subsequent analysis.
(4) statistical analysis is carried out using SPSS18.0, the enumeration data between test rating and clinical data uses
Pearson Chi-square Tests, measurement data are examined using t.The analysis of Testing index and clinical prognosis is given birth to using KaPlan-Meier
Analysis is deposited, the difference of survivorship curve is compared in logarithm rank test (log-ranktest).The present invention shows TCP1 albumen and mammary gland
The prognosis of cancer has significant correlation, to predict that the relapse and metastasis of breast cancer and postoperative survival rate provide a brand-new way
Footpath, the prognosis to patient with breast cancer play an important role.When the scoring of cancerous tissue TCP1 SABCs is higher than 0.36, breast cancer easily goes out
Existing relapse and metastasis, patient with breast cancer are postoperative easily dead.
The beneficial effects are mainly as follows：Present invention research finds TCP1 expression and patient's art in breast cancer
Afterwards there is significant relation in prognosis, imply that TCP1 can be as effective early warning albumen of the Postoperative determination of breast cancer.Therefore, it is of the invention
Application of the TCP1 albumen in Mammary cancer prognosis evaluation reagent kit is prepared is provided, prompts the albumen to be used to prepare judgement
The protein molecular marker of patient with breast cancer's prognosis, also there is important finger for the postoperative monitoring of breast cancer patients and sequential therapy
Lead meaning.
Brief description of the drawings
Fig. 1 be TCP1 in postoperative 1 year of patient with breast cancer without the tissue samples of relapse and metastasis in weak expression；
Fig. 2 is TCP1 strongly expresseds in the tissue samples of relapse and metastasis in postoperative 1 year of patient with breast cancer；
Fig. 3 is TCP1 weak expressions in mammary gland cancer beside organism；
Fig. 4 is TCP1 low expressions group and high expression group survivorship curve in breast cancer tissue.
Embodiment
With reference to specific embodiment, the present invention is described further, but protection scope of the present invention is not limited to
This：
Embodiment 1
For the kit of Mammary cancer prognosis evaluation, the kit includes：For to the TCP1 in breast cancer tissue
People source TCP1 monoclonal antibodies or TCP1 polyclonal antibodies that the relative expression quantity of albumen is detected, immunohistochemical experiment examination
Agent.Wherein, immunohistochemical experiment reagent includes：Dimethylbenzene, ethanol, 3%H2O2, 3%BSA confining liquids, DAB colour reagents, bush
Element, horseradish peroxidase, PBS and 0.01M EDTA repair liquid.
People source TCP1 polyclonal antibodies of the present invention are the TCP1 protein immunization rabbits shown in SEQ ID NO.1 as sequence
Obtain.It can voluntarily prepare, can also use commodity purchased in market.
Immunohistochemical experiment reagent of the present invention includes：Dimethylbenzene, ethanol, 3%H2O2(aqueous solution), 3%BSA are closed
Liquid (being prepared with PBS), DAB colour reagents, haematoxylin, horseradish peroxidase (be used for mark secondary antibody), PBS (pH7.4),
0.01M EDTA repair liquid.
Embodiment 2
Application of the TCP1 albumen in Mammary cancer prognosis evaluation reagent kit is prepared, the application are to utilize reagent of the present invention
Box carries out Mammary cancer prognosis evaluation, and method is as follows：
(1) patient with breast cancer's Pathologic specimen is taken, using people source TCP1 monoclonal antibodies or people source TCP1 polyclonal antibodies, with
And immunohistochemical experiment reagent, carry out immunohistochemical staining；
Wherein, Pathologic specimen comes from the pathologic sampling in patient with breast cancer's biopsy or art, postoperative.SABC side
Method utilizes SP decoration methods, comprises the following steps that：
(a) breast cancer tissue's paraffin section is prepared, 60 DEG C of baking boxs are stayed overnight.
(b) section dewaxing.Soak successively：Dimethylbenzene I：10min；Dimethylbenzene II：10min；Dimethylbenzene III：10min.
(c) section aquation.Soak successively：Absolute ethyl alcohol：3min；90% (v/v) ethanol：3min；80% ethanol：3min；
75% ethanol：3min.
(d) PBS 3 times, each 5min.
(e) EDTA antigens Pressure method：Section is put into 0.01M EDTA and repairs liquid immersion, boiling water bath 5min, is cooled to room
Temperature.PBS 3 times, each 5min.
(f) 300 μ L 3% (w/w) aqueous hydrogen peroxide solution, 37 DEG C of 10min are added.PBS 3 times, each 5min.
(g) 300 μ L 3% (w/w) BSA confining liquids (PBS preparations), 37 DEG C of 1h are added.PBS 3 times, each 5min.
(h) primary antibody is added：TCP1 antibody concentrations：1：500,4 DEG C of refrigerators take out after placing 16h, room temperature rewarming 15min, so
PBS is washed 4 times afterwards, each 5min.
(i) secondary antibody is added dropwise, described secondary antibody is that horseradish peroxidase-labeled goat anti-rabbit igg (steps novel agent purchased from Foochow
Company, instant, without dilution), 37 DEG C of 45min.PBS is washed 4 times, each 5min.
(j) PBS is washed 3 times, each 5min.Develop the color DAB (DAB colour reagent boxes, work being given birth to purchased from Shanghai) 2-10min, under mirror
Observation；Distilled water, which is washed, only to develop the color, and haematoxylin redyes 10s, is rinsed and soaked with running water.
(k) it is dehydrated.Soak successively：75% ethanol：2min；80% ethanol：2min；90% ethanol：2min；Absolute ethyl alcohol：
2min。
(L) electricity consumption blowing drying, neutral gum, cover glass covering are added.
(2) 3 visual field shootings of breast cancer tissue and cancer beside organism are randomly selected using microscope and imaging device, utilized
Aperio Image Scope softwares are scanned to the photograph of tissue samples, and the Algorithms of the software is used after scanning
(Positive Pixel Count V9) program carries out positive strength calculating to each sample, and it is as follows to calculate data：
(3) the immunohistochemistry score calculation of each tissue samples is Positivity × Log10 [255/Iavg], its
Middle Positivity=NPositive/NTotal, i.e. positive rate, computational methods are positive pixels quantity/colour developing total quantity；
Iavg=(Iwp+Ip+Isp)/(Nwp+Np+Nsp), i.e., positive mean intensity, computational methods are positive mean intensity=(weak sun
Property pixel overall strength+positive pixels overall strength+strong positive pixel overall strength)/(weakly positive pixel number+positive pixels quantity+strong
Positive pixels quantity), the as SABC scoring of the tissue, for subsequent analysis.The high low expression standards of TCP1 are with 326 breasts
The median (0.36) of TCP1 expression scoring is boundary in adenocarcinoma tissue.
(4) statistical analysis is carried out using SPSS18.0, the enumeration data between test rating and clinical data uses
Pearson Chi-square Tests, measurement data are examined using t.The analysis of Testing index and clinical prognosis is given birth to using KaPlan-Meier
Analysis is deposited, the difference of survivorship curve is compared in logarithm rank test (log-ranktest).
According to the method described above, testing result is as Figure 1-3 in the tumor tissues of 326 breast cancer patients by the present invention：
TCP1 weak expressions in non-relapse and metastasis group in 1 year；The high expression (Fig. 2) in relapse and metastasis group；The weak expression in cancer beside organism
(Fig. 3).
TCP1 and the relation of patient with breast cancer's prognosis：
Analyzed by Kaplan-Meier survivorship curves, TCP1 expression degree is related to the prognosis of patient with breast cancer, such as
Shown in Fig. 4, patient's mean survival time (MST) (survival rate) of the high expression of TCP1, is less than the patient of TCP1 low expressions.
Embodiment 3
Take certain Mammary cancer tumor sample to carry out specimens paraffin embedding slices, and utilize above-described immunohistochemistry side
Method is detected, and is computed, and the TCP1 SABCs tissue scoring of its cancerous tissue is 0.586.Found by Follow-up After, the trouble
Breast cancer relapse transfer, postoperative death in 15 months occur for 9th month after surgery for person.
Embodiment 4
Take certain Mammary cancer tumor sample to carry out specimens paraffin embedding slices, and utilize above-described immunohistochemistry side
Method is detected, and is computed, and the TCP1 SABCs tissue scoring of its cancerous tissue is 0.128.Found by Follow-up After, the trouble
Person 1 does not have found transfer and relapse every year after surgery, is still living and in good health.
By the above results showed that TCP1 protein molecular relative expression quantity energy is detected by using the method for SABC
Predict breast cancer DISTANT METASTASES IN risk and the postoperative existence or death of patient.When cancerous tissue TCP1 SABC scoring is higher than
When 0.36, easily there is relapse and metastasis in breast cancer, and patient with breast cancer is postoperative easily dead.Obvious TCP1 albumen has related to breast cancer
Property, therefore, its expression quantity is carried out using TCP1 albumen as protein molecular marker to recur after detection can predict mammary cancer surgery
The events such as transfer, and judging prognosis.
Sequence table
<110>Fujian Normal University
<120>Application of the TCP1 albumen in Mammary cancer prognosis evaluation reagent kit is prepared, Prognosis in Breast Cancer assess kit
And method
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<170> SIPOSequenceListing 1.0
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Met Glu Gly Pro Leu Ser Val Phe Gly Asp Arg Ser Thr Gly Glu Thr
1 5 10 15
Ile Arg Ser Gln Asn Val Met Ala Ala Ala Ser Ile Ala Asn Ile Val
20 25 30
Lys Ser Ser Leu Gly Pro Val Gly Leu Asp Lys Met Leu Val Asp Asp
35 40 45
Ile Gly Asp Val Thr Ile Thr Asn Asp Gly Ala Thr Ile Leu Lys Leu
50 55 60
Leu Glu Val Glu His Pro Ala Ala Lys Val Leu Cys Glu Leu Ala Asp
65 70 75 80
Leu Gln Asp Lys Glu Val Gly Asp Gly Thr Thr Ser Val Val Ile Ile
85 90 95
Ala Ala Glu Leu Leu Lys Asn Ala Asp Glu Leu Val Lys Gln Lys Ile
100 105 110
His Pro Thr Ser Val Ile Ser Gly Tyr Arg Leu Ala Cys Lys Glu Ala
115 120 125
Val Arg Tyr Ile Asn Glu Asn Leu Ile Val Asn Thr Asp Glu Leu Gly
130 135 140
Arg Asp Cys Leu Ile Asn Ala Ala Lys Thr Ser Met Ser Ser Lys Ile
145 150 155 160
Ile Gly Ile Asn Gly Asp Phe Phe Ala Asn Met Val Val Asp Ala Val
165 170 175
Leu Ala Ile Lys Tyr Thr Asp Ile Arg Gly Gln Pro Arg Tyr Pro Val
180 185 190
Asn Ser Val Asn Ile Leu Lys Ala His Gly Arg Ser Gln Met Glu Ser
195 200 205
Met Leu Ile Ser Gly Tyr Ala Leu Asn Cys Val Val Gly Ser Gln Gly
210 215 220
Met Pro Lys Arg Ile Val Asn Ala Lys Ile Ala Cys Leu Asp Phe Ser
225 230 235 240
Leu Gln Lys Thr Lys Met Lys Leu Gly Val Gln Val Val Ile Thr Asp
245 250 255
Pro Glu Lys Leu Asp Gln Ile Arg Gln Arg Glu Ser Asp Ile Thr Lys
260 265 270
Glu Arg Ile Gln Lys Ile Leu Ala Thr Gly Ala Asn Val Ile Leu Thr
275 280 285
Thr Gly Gly Ile Asp Asp Met Cys Leu Lys Tyr Phe Val Glu Ala Gly
290 295 300
Ala Met Ala Val Arg Arg Val Leu Lys Arg Asp Leu Lys Arg Ile Ala
305 310 315 320
Lys Ala Ser Gly Ala Thr Ile Leu Ser Thr Leu Ala Asn Leu Glu Gly
325 330 335
Glu Glu Thr Phe Glu Ala Ala Met Leu Gly Gln Ala Glu Glu Val Val
340 345 350
Gln Glu Arg Ile Cys Asp Asp Glu Leu Ile Leu Ile Lys Asn Thr Lys
355 360 365
Ala Arg Thr Ser Ala Ser Ile Ile Leu Arg Gly Ala Asn Asp Phe Met
370 375 380
Cys Asp Glu Met Glu Arg Ser Leu His Asp Ala Leu Cys Val Val Lys
385 390 395 400
Arg Val Leu Glu Ser Lys Ser Val Val Pro Gly Gly Gly Ala Val Glu
405 410 415
Ala Ala Leu Ser Ile Tyr Leu Glu Asn Tyr Ala Thr Ser Met Gly Ser
420 425 430
Arg Glu Gln Leu Ala Ile Ala Glu Phe Ala Arg Ser Leu Leu Val Ile
435 440 445
Pro Asn Thr Leu Ala Val Asn Ala Ala Gln Asp Ser Thr Asp Leu Val
450 455 460
Ala Lys Leu Arg Ala Phe His Asn Glu Ala Gln Val Asn Pro Glu Arg
465 470 475 480
Lys Asn Leu Lys Trp Ile Gly Leu Asp Leu Ser Asn Gly Lys Pro Arg
485 490 495
Asp Asn Lys Gln Ala Gly Val Phe Glu Pro Thr Ile Val Lys Val Lys
500 505 510
Ser Leu Lys Phe Ala Thr Glu Ala Ala Ile Thr Ile Leu Arg Ile Asp
515 520 525
Asp Leu Ile Lys Leu His Pro Glu Ser Lys Asp Asp Lys His Gly Ser
530 535 540
Tyr Glu Asp Ala Val His Ser Gly Ala Leu Asn Asp
545 550 555
Claims (6)
- Application of the 1.TCP1 albumen in Mammary cancer prognosis evaluation reagent kit is prepared.
- 2. application of the TCP1 albumen according to claim 1 in Mammary cancer prognosis evaluation reagent kit is prepared, it is special Sign is：Using TCP1 albumen as molecular labeling, using people source TCP1 monoclonal antibodies or people source TCP1 polyclonal antibodies, with reference to Immunohistochemical experiment reagent, relative expression quantity of the detection TCP1 albumen in breast cancer tissue.
- 3. application of the TCP1 albumen according to claim 2 in Mammary cancer prognosis evaluation reagent kit is prepared, it is special Sign is：The people source TCP1 polyclonal antibodies are obtained as sequence for the TCP1 protein immunizations rabbit shown in SEQ ID NO.1.
- 4. the kit applied to Mammary cancer prognosis evaluation, it is characterised in that：The kit includes being used for breast cancer The people source TCP1 monoclonal antibodies or people source TCP1 polyclonal antibodies that the relative expression quantity of TCP1 albumen in tissue is detected, And immunohistochemical experiment reagent.
- 5. according to claim 4 be applied to Mammary cancer prognosis evaluation reagent kit, it is characterised in that：The immune group Changing experiment reagent includes：Dimethylbenzene, ethanol, 3%H2O2, 3%BSA confining liquids, DAB colour reagents, haematoxylin, horseradish peroxidase Enzyme, PBS and 0.01M EDTA repair liquid.
- 6. a kind of method that kit based on described in claim 4 carries out Mammary cancer prognosis evaluation, it is characterised in that：Its Comprise the following steps：（1）Patient with breast cancer's Pathologic specimen is taken, using people source TCP1 monoclonal antibodies or people source TCP1 polyclonal antibodies, and is exempted from Epidemic disease group experiment reagent, carry out immunohistochemical staining；（2）3 visuals field of breast cancer tissue and cancer beside organism, which are randomly selected, using microscope and imaging device is shot for digital photograph；（3）Tissue samples are scanned using Aperio Image Scope softwares, using the software after scanning Algorithms programs carry out positive strength calculating to each sample, are scaled the immunohistochemistry scoring of the sample；（4）The postoperative prognosis situation of patient is assessed by resulting scoring.
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| CN109342742A (en) * | 2018-11-28 | 2019-02-15 | 中山大学孙逸仙纪念医院 | A kind of kit for Prognosis in Breast Cancer judgement |
| CN109507161A (en) * | 2018-11-28 | 2019-03-22 | 中山大学孙逸仙纪念医院 | Application of the TIMP-1 in the kit and suppression cancer drug of preparation Prognosis in Breast Cancer diagnosis |
| CN110286228A (en) * | 2019-04-16 | 2019-09-27 | 福建师范大学 | Nuclear lamina protein A is preparing the application in clear cell carcinoma of kidney Postoperative determination assessment kit |
| CN110412295A (en) * | 2019-08-09 | 2019-11-05 | 中国人民解放军军事科学院军事医学研究院 | PTEN Nedd8 modifies the invention and application as breast cancer Novel marker and its specific antibody |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109342742A (en) * | 2018-11-28 | 2019-02-15 | 中山大学孙逸仙纪念医院 | A kind of kit for Prognosis in Breast Cancer judgement |
| CN109507161A (en) * | 2018-11-28 | 2019-03-22 | 中山大学孙逸仙纪念医院 | Application of the TIMP-1 in the kit and suppression cancer drug of preparation Prognosis in Breast Cancer diagnosis |
| CN110286228A (en) * | 2019-04-16 | 2019-09-27 | 福建师范大学 | Nuclear lamina protein A is preparing the application in clear cell carcinoma of kidney Postoperative determination assessment kit |
| CN110412295A (en) * | 2019-08-09 | 2019-11-05 | 中国人民解放军军事科学院军事医学研究院 | PTEN Nedd8 modifies the invention and application as breast cancer Novel marker and its specific antibody |
| CN110412295B (en) * | 2019-08-09 | 2022-09-13 | 中国人民解放军军事科学院军事医学研究院 | PTEN Nedd8 modified breast cancer novel marker and invention and application of specific antibody thereof |
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