CN110018317A - Application of the nuclear lamina protein A in preparation cutaneum carcinoma Postoperative determination assessment kit - Google Patents

Application of the nuclear lamina protein A in preparation cutaneum carcinoma Postoperative determination assessment kit Download PDF

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CN110018317A
CN110018317A CN201910305485.8A CN201910305485A CN110018317A CN 110018317 A CN110018317 A CN 110018317A CN 201910305485 A CN201910305485 A CN 201910305485A CN 110018317 A CN110018317 A CN 110018317A
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cutaneum carcinoma
nuclear lamina
lamina protein
glu
leu
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王清水
林尧
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Fujian Normal University
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Fujian Normal University
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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
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
    • G01N33/57407—Specifically defined cancers
    • G01N33/5743—Specifically defined cancers of skin, e.g. melanoma
    • 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
    • G01N33/6881—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids from skin
    • 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
    • G01N33/6893—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving proteins, peptides or amino acids related to diseases not provided for elsewhere
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/50—Determining the risk of developing a disease
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/52—Predicting or monitoring the response to treatment, e.g. for selection of therapy based on assay results in personalised medicine; Prognosis
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N2800/00—Detection or diagnosis of diseases
    • G01N2800/54—Determining the risk of relapse

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Immunology (AREA)
  • Molecular Biology (AREA)
  • Chemical & Material Sciences (AREA)
  • Biomedical Technology (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
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  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Oncology (AREA)
  • Hospice & Palliative Care (AREA)
  • Investigating Or Analysing Biological Materials (AREA)

Abstract

The present invention provides new opplication of the nuclear lamina protein A in preparation cutaneum carcinoma Postoperative determination assessment kit.The present inventor after extensive and in-depth study, has found for the first time, using relative expression quantity of the ImmunohistochemistryMethods Methods detection nuclear lamina protein A in cutaneous carcinoma tissue, can judge that the risk of cutaneum carcinoma relapse and metastasis occurs in cutaneum carcinoma patient.The beneficial effects are mainly reflected as follows: the present invention provides application of the nuclear lamina protein A in preparation cutaneum carcinoma Postoperative determination assessment kit, prompt the albumen that can be used to prepare the protein molecular marker for judging cutaneum carcinoma patient's prognosis, monitoring postoperative for cutaneum carcinoma patient and sequential therapy also have important directive significance.

Description

Application of the nuclear lamina protein A in preparation cutaneum carcinoma Postoperative determination assessment kit
Technical field
The present invention relates to application of the nuclear lamina protein A in preparation cutaneum carcinoma Postoperative determination assessment kit.
Background technique
Cutaneum carcinoma, that is, malignant tumour of skin has different names according to the source of tumour cell difference, including epidermis, skin Skin appendicle, skin soft tissue, peripheral nerve, melanocyte, skin lymphoreticular tissue and hematopoietic tissue etc..Some It is to occur to be transferred to the metastatic tumo(u)r of skin in its hetero-organization.
The occurrence and development and prognosis of cutaneum carcinoma and the activation of oncogene and tumor suppressor gene functionally inactive close association, be mostly because Element induction, the complicated pathologic process that polygenes participates in.In recent years, the molecular mechanism of cutaneum carcinoma early stage relapse and metastasis is the field Heat subject deeply illustrates the molecular mechanism, and imports magnetic target therapy measure in its specific link, is expected to very significantly Improve the postoperative overall treatment effect of cutaneum carcinoma.Therefore, it is related to seek effective cutaneum carcinoma skin excision early postoperation recurrence Early warning molecular marker illustrates the relationship of itself and cutaneum carcinoma early stage relapse and metastasis, this is to raising cutaneum carcinoma aftertreatment effect, assessment Cutaneum carcinoma early stage risk of recurrence, judging prognosis and individualized treatment have vital meaning.
Nuclear lamina protein A is two albumen encoded by lamin, participates in the tissue of nucleus nuclear membrane, influences gene Group stability simultaneously has an impact cell differentiation.Nuclear lamina protein A abnormal expression is generally existing in human tumor, and mutation causes A variety of lamin diseases, such as Emery-Dreifuss muscular dystrophy, dilated cardiomyopathy and hutchinson-Gilford syndrome.
The research of the invention finds that there is significant close with patient's Postoperative determination in the nuclear lamina protein A expression in cutaneum carcinoma System implies that nuclear lamina protein A can be used as effective early warning albumen of Postoperative determination of cutaneum carcinoma.
Cutaneum carcinoma threatens one of maximum tumour as to human health, and the molecular mechanism of its generation is still unclear so far, Also lack the molecular target of specificity to its treatment, and nuclear lamina protein A is as highly important tumour oncogene, at present still No document report nuclear lamina protein A is related to cutaneum carcinoma prognosis or skin metastasis of cancer is judged.
Summary of the invention
It is an object of the present invention to provide new opplication of the nuclear lamina protein A in preparation cutaneum carcinoma Postoperative determination assessment kit.
The technical solution adopted by the present invention is that:
Application of the nuclear lamina protein A in preparation cutaneum carcinoma Postoperative determination assessment kit.
The present inventor after extensive and in-depth study, has found for the first time, detects nuclear lamina protein A using ImmunohistochemistryMethods Methods Relative expression quantity in cutaneous carcinoma tissue can judge that the risk of cutaneum carcinoma relapse and metastasis occurs in cutaneum carcinoma patient.Based on core The correlation of lamina protein A expression quantity and cutaneum carcinoma relapse and metastasis carries out its expression quantity using the albumen as prognostic markers object Detection can be used for instructing the Index for diagnosis of cutaneum carcinoma, therefore can utilize nuclear fabric layer protein using nuclear lamina protein A as molecular labeling White A monoclonal antibody or polyclonal antibody detect nuclear lamina protein A in cutaneous carcinoma tissue in conjunction with immunohistochemical experiment reagent Relative expression quantity.
The kit specifically includes that source of people nuclear lamina protein A monoclonal antibody or polyclonal antibody, immunohistochemical experiment Reagent.The immunohistochemical experiment reagent is the common agents in the immunohistochemical experiment of this field.
Inventor is having found that nuclear lamina protein A is expressed in relapse and metastasis cutaneous carcinoma tissue is higher than non-relapse and metastasis skin Cancerous tissue can speculate that nuclear lamina protein A plays a significant role in the transfer of cutaneum carcinoma postoperative recurrence.Consult domestic and foreign literature, core The generation of lamina protein A and cutaneum carcinoma and the correlative study of relapse and metastasis are few, and in this experiment, nuclear lamina protein A is in skin Expression up-regulation in cancerous tissue, and downward is then expressed by cancer and in normal skin tissue, and compared with non-relapse and metastasis group, core Lamina protein A is expressed in relapse and metastasis group and is also raised, and shows that nuclear lamina protein A may participate in cutaneum carcinoma hair as promotive factor Raw development process.It is analyzed by Kaplan-Meier survivorship curve, nuclear lamina protein A expresses the prognosis of degree and cutaneum carcinoma patient It is related, the highly expressed patient's prognosis mala (P < 0.05) of nuclear lamina protein A.
In conclusion nuclear lamina protein A high expression in cutaneous carcinoma tissue, the high expression of nuclear lamina protein A and cutaneum carcinoma The transfer of patient's postoperative recurrence is related.Nuclear lamina protein A can be used as an important candidate molecular marker object of cutaneum carcinoma prognosis.
Preferably, the source of people nuclear lamina protein A polyclonal antibody is nuclear fabric layer protein shown in SEQ ID NO.1 as sequence White A immune rabbit obtains, and can voluntarily prepare, commercially available commodity can also be used.
Specifically, the immunohistochemical experiment reagent includes: dimethylbenzene, ethyl alcohol, 3%H2O2(aqueous solution), 3%BSA closing Liquid (being prepared with PBS), DAB colour reagent, haematoxylin, horseradish peroxidase (for marking secondary antibody), PBS (pH7.4), 0.01M EDTA repairs liquid.
The application method of kit of the present invention is as follows:
(a) pathologic sampling of the Pathologic specimen in cutaneum carcinoma patient biopsy or art, postoperative.
(b) ImmunohistochemistryMethods Methods utilize SP decoration method, the specific steps are as follows:
(c) cutaneous carcinoma tissue paraffin section is prepared, 60 DEG C of ovens are stayed overnight.
(d) it is sliced dewaxing.It successively impregnates: dimethylbenzene I:10min；Dimethylbenzene II:10min；Dimethylbenzene III:10min.
(e) it is sliced aquation.It successively impregnates: dehydrated alcohol: 3min；90% (v/v) ethyl alcohol: 3min；80% ethyl alcohol: 3min； 75% ethyl alcohol: 3min.
(f) PBS is cleaned 3 times, each 5min.
(h) EDTA antigen Pressure method: slice is put into 0.01M EDTA and repairs liquid immersion, and boiling water bath 5min is cooled to room Temperature.PBS is cleaned 3 times, each 5min.
(I) 3% (w/w) aqueous hydrogen peroxide solution of 300 μ L, 37 DEG C of 10min are added.PBS is cleaned 3 times, each 5min.
(J) 3% (w/w) the BSA confining liquid (PBS preparation) of 300 μ L, 37 DEG C of 1h are added.PBS is cleaned 3 times, each 5min.
(K) primary antibody: nuclear lamina protein A antibody concentration: 1:500 is added, 4 DEG C of refrigerators take out after placing 16h, room temperature rewarming 15min, then PBS is washed 4 times, each 5min.
(L) secondary antibody is added dropwise, the 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.
(M) PBS is washed 3 times, each 5min.Develop the color DAB (DAB colour reagent box, purchased from the raw work in Shanghai) 2-10min, under mirror Observation；Distilled water, which is washed, only to develop the color, and haematoxylin redyes 10s, is rinsed and is impregnated with tap water.
(N) it is dehydrated.It successively impregnates: 75% ethyl alcohol: 2min；80% ethyl alcohol: 2min；90% ethyl alcohol: 2min；Dehydrated alcohol: 2min。
(O) neutral gum, coverslip covering is added in electricity consumption blowing drying.
(P) 3 visuals field of cutaneous carcinoma tissue and cancer beside organism are randomly selected using microscope and imaging device to shoot, is utilized Aperio Image Scope software is scanned 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:
(Q) the immunohistochemistry scoring of each tissue samples is calculated as Positivity × Log10 [255/Iavg], Middle Positivity=NPositive/NTotal, i.e. positive rate, calculation method are positive pixels quantity/colour developing total quantity； Iavg=(Iwp+Ip+Isp)/(Nwp+Np+Nsp), i.e., positive mean intensity, calculation method 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 immunohistochemistry scoring of the tissue is used for subsequent analysis.
(L) for statistical analysis using SPSS18.0, the enumeration data between test rating and clinical data uses Pearson Chi-square Test, measurement data are examined using t.The analysis of Testing index and clinical prognosis is raw using KaPlan-Meier Analysis is deposited, logarithm rank sum test (log-ranktest) compares the difference of survivorship curve.The invention shows nuclear lamina protein As and skin The prognosis of skin cancer has significant correlation, to predict that relapse and metastasis and the postoperative survival rate of cutaneum carcinoma provide a completely new way Diameter plays an important role to the prognosis of cutaneum carcinoma patient.When the scoring of cancerous tissue nuclear lamina protein A immunohistochemistry is higher than 0.586, skin Easily there is relapse and metastasis, the postoperative easy death of cutaneum carcinoma patient in skin cancer.
The beneficial effects are mainly reflected as follows: it is postoperative pre- in preparation cutaneum carcinoma that the present invention provides nuclear lamina protein As The application in kit is assessed afterwards, prompts the albumen that can be used to prepare the protein molecular marker for judging cutaneum carcinoma patient's prognosis, Monitoring postoperative for cutaneum carcinoma patient and sequential therapy also have important directive significance.
Detailed description of the invention
Fig. 1 is nuclear lamina protein A expression in the tissue samples of the postoperative no relapse and metastasis of cutaneum carcinoma patient；
Fig. 2 is nuclear lamina protein A expression in the tissue samples that cutaneum carcinoma patient's postoperative recurrence shifts；
Fig. 3 is nuclear lamina protein A low expression group and high expression group survivorship curve in cutaneous carcinoma tissue；
Specific embodiment
The present invention is described further combined with specific embodiments below, but protection scope of the present invention is not limited in This:
Embodiment 1:
(a) pathologic sampling of the Pathologic specimen in cutaneum carcinoma patient biopsy or art, postoperative.
(b) ImmunohistochemistryMethods Methods utilize SP decoration method, the specific steps are as follows:
(c) cutaneous carcinoma tissue paraffin section is prepared, 60 DEG C of ovens are stayed overnight.
(d) it is sliced dewaxing.It successively impregnates: dimethylbenzene I:10min；Dimethylbenzene II:10min；Dimethylbenzene III:10min.
(e) it is sliced aquation.It successively impregnates: dehydrated alcohol: 3min；90% (v/v) ethyl alcohol: 3min；80% ethyl alcohol: 3min； 75% ethyl alcohol: 3min.
(f) PBS is cleaned 3 times, each 5min.
(h) EDTA antigen Pressure method: slice is put into 0.01M EDTA and repairs liquid immersion, and boiling water bath 5min is cooled to room Temperature.PBS is cleaned 3 times, each 5min.
(I) 3% (w/w) aqueous hydrogen peroxide solution of 300 μ L, 37 DEG C of 10min are added.PBS is cleaned 3 times, each 5min.
(J) 3% (w/w) the BSA confining liquid (PBS preparation) of 300 μ L, 37 DEG C of 1h are added.PBS is cleaned 3 times, each 5min.
(K) primary antibody: nuclear lamina protein A antibody concentration: 1:500 is added, 4 DEG C of refrigerators take out after placing 16h, room temperature rewarming 15min.PBS is washed 4 times, each 5min.
(L) secondary antibody is added dropwise, the 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.
(M) PBS is washed 3 times, each 5min.Develop the color DAB (DAB colour reagent box, purchased from the raw work in Shanghai) 2-10min, under mirror Observation；Distilled water, which is washed, only to develop the color, and haematoxylin redyes 10s, is rinsed and is impregnated with tap water.
(N) it is dehydrated.It successively impregnates: 75% ethyl alcohol: 2min；80% ethyl alcohol: 2min；90% ethyl alcohol: 2min；Dehydrated alcohol: 2min。
(O) neutral gum, coverslip covering is added in electricity consumption blowing drying.
(P) 3 visuals field of cutaneous carcinoma tissue and cancer beside organism are randomly selected using microscope and imaging device to shoot, is utilized Aperio Image Scope software is scanned the photo 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:
(Q) the immunohistochemistry scoring of each tissue samples is calculated as Positivity × Log10 [255/Iavg], Middle Positivity=NPositive/NTotal, i.e. positive rate, calculation method are positive pixels quantity/colour developing total quantity； Iavg=(Iwp+Ip+Isp)/(Nwp+Np+Nsp), i.e., positive mean intensity, calculation method 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 immunohistochemistry scoring of the tissue is used for subsequent analysis.The high low expression standard of nuclear lamina protein A with The median (0.586) of nuclear lamina protein A expression scoring is boundary in 30 cutaneous carcinoma tissues.
(L) for statistical analysis using SPSS18.0, the enumeration data between test rating and clinical data uses Pearson Chi-square Test, measurement data are examined using t.The analysis of Testing index and clinical prognosis is raw using KaPlan-Meier Analysis is deposited, logarithm rank sum test (log-ranktest) compares the difference of survivorship curve.
According to the method described above, present invention testing result in the tumor tissues of 30 cutaneum carcinoma patients is as shown in Figs. 1-2: core Lamina protein A is lower than relapse and metastasis group (Fig. 2) without the expression (Fig. 1) in relapse and metastasis group.
The relationship of nuclear lamina protein A and cutaneum carcinoma patient prognosis:
It is analyzed by Kaplan-Meier survivorship curve, the expression degree of nuclear lamina protein A and the prognosis of cutaneum carcinoma patient Related (Fig. 3).
Embodiment 2:
It takes the postoperative tumor sample of certain cutaneum carcinoma to carry out specimens paraffin embedding slices, and utilizes above-described immunohistochemistry side Method is detected, and is computed, and the nuclear lamina protein A immunohistochemistry tissue scoring of cancerous tissue is 0.732.It is sent out by Follow-up After It is existing, 20th month after surgery generation cutaneum carcinoma relapse and metastasis of the patient, postoperative death in 42 months.
Embodiment 3:
It takes the postoperative tumor sample of certain cutaneum carcinoma to carry out specimens paraffin embedding slices, and utilizes above-described immunohistochemistry side Method is detected, and is computed, and the nuclear lamina protein A immunohistochemistry tissue scoring of cancerous tissue is 0.294.It is sent out by Follow-up After Existing, which 3 does not find transfer and relapse every year after surgery, is still living and in good health.
By the above test result it is found that the method by using immunohistochemistry detects nuclear lamina protein A molecule relative expression Amount can predict cutaneum carcinoma DISTANT METASTASES IN risk and the postoperative existence or death of patient.When being immunized for cancerous tissue nuclear lamina protein A When groupization scoring is higher than 0.586, easily there is relapse and metastasis, the postoperative easy death of cutaneum carcinoma patient in cutaneum carcinoma.Obvious nuclear lamina protein A There is correlation with cutaneum carcinoma, therefore, carrying out detection to its expression quantity using nuclear lamina protein A as protein molecular marker can be pre- Survey the events such as cutaneum carcinoma recurrence after operation transfer, and judging prognosis.
The basic principles, main features and advantages of the present invention have been shown and described above.The technology of the industry Personnel are it should be appreciated that the present invention is not limited to the above embodiments, and the above embodiments and description only describe this The principle of invention, various changes and improvements may be made to the invention without departing from the spirit and scope of the present invention, these changes Change and improvement all fall within the protetion scope of the claimed invention.The claimed scope of the invention by appended claims and its Equivalent defines.
Sequence table
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<120>application of the nuclear lamina protein A in preparation cutaneum carcinoma Postoperative determination assessment kit
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Met Glu Thr Pro Ser Gln Arg Arg Ala Thr Arg Ser Gly Ala Gln Ala
1 5 10 15
Ser Ser Thr Pro Leu Ser Pro Thr Arg Ile Thr Arg Leu Gln Glu Lys
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Glu Asp Leu Gln Glu Leu Asn Asp Arg Leu Ala Val Tyr Ile Asp Arg
35 40 45
Val Arg Ser Leu Glu Thr Glu Asn Ala Gly Leu Arg Leu Arg Ile Thr
50 55 60
Glu Ser Glu Glu Val Val Ser Arg Glu Val Ser Gly Ile Lys Ala Ala
65 70 75 80
Tyr Glu Ala Glu Leu Gly Asp Ala Arg Lys Thr Leu Asp Ser Val Ala
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Lys Glu Arg Ala Arg Leu Gln Leu Glu Leu Ser Lys Val Arg Glu Glu
100 105 110
Phe Lys Glu Leu Lys Ala Arg Asn Thr Lys Lys Glu Gly Asp Leu Ile
115 120 125
Ala Ala Gln Ala Arg Leu Lys Asp Leu Glu Ala Leu Leu Asn Ser Lys
130 135 140
Glu Ala Ala Leu Ser Thr Ala Leu Ser Glu Lys Arg Thr Leu Glu Gly
145 150 155 160
Glu Leu His Asp Leu Arg Gly Gln Val Ala Lys Leu Glu Ala Ala Leu
165 170 175
Gly Glu Ala Lys Lys Gln Leu Gln Asp Glu Met Leu Arg Arg Val Gly
180 185 190
Ala Glu Asn Arg Leu Gln Thr Met Lys Glu Glu Leu Asp Phe Gln Lys
195 200 205
Asn Ile Tyr Ser Glu Glu Leu Arg Glu Thr Lys Arg Arg His Glu Thr
210 215 220
Arg Leu Val Glu Ile Asp Asn Gly Lys Gln Arg Glu Phe Glu Ser Arg
225 230 235 240
Leu Ala Asp Ala Leu Gln Glu Leu Arg Ala Gln His Glu Asp Gln Val
245 250 255
Glu Gln Tyr Lys Lys Glu Leu Glu Lys Thr Tyr Ser Ala Lys Leu Asp
260 265 270
Asn Ala Arg Gln Ser Ala Glu Arg Asn Ser Asn Leu Val Gly Ala Ala
275 280 285
His Glu Glu Leu Gln Gln Ser Arg Ile Arg Ile Asp Ser Leu Ser Ala
290 295 300
Gln Leu Ser Gln Leu Gln Lys Gln Leu Ala Ala Lys Glu Ala Lys Leu
305 310 315 320
Arg Asp Leu Glu Asp Ser Leu Ala Arg Glu Arg Asp Thr Ser Arg Arg
325 330 335
Leu Leu Ala Glu Lys Glu Arg Glu Met Ala Glu Met Arg Ala Arg Met
340 345 350
Gln Gln Gln Leu Asp Glu Tyr Gln Glu Leu Leu Asp Ile Lys Leu Ala
355 360 365
Leu Asp Met Glu Ile His Ala Tyr Arg Lys Leu Leu Glu Gly Glu Glu
370 375 380
Glu Arg Leu Arg Leu Ser Pro Ser Pro Thr Ser Gln Arg Ser Arg Gly
385 390 395 400
Arg Ala Ser Ser His Ser Ser Gln Thr Gln Gly Gly Gly Ser Val Thr
405 410 415
Lys Lys Arg Lys Leu Glu Ser Thr Glu Ser Arg Ser Ser Phe Ser Gln
420 425 430
His Ala Arg Thr Ser Gly Arg Val Ala Val Glu Glu Val Asp Glu Glu
435 440 445
Gly Lys Phe Val Arg Leu Arg Asn Lys Ser Asn Glu Asp Gln Ser Met
450 455 460
Gly Asn Trp Gln Ile Lys Arg Gln Asn Gly Asp Asp Pro Leu Leu Thr
465 470 475 480
Tyr Arg Phe Pro Pro Lys Phe Thr Leu Lys Ala Gly Gln Val Val Thr
485 490 495
Ile Trp Ala Ala Gly Ala Gly Ala Thr His Ser Pro Pro Thr Asp Leu
500 505 510
Val Trp Lys Ala Gln Asn Thr Trp Gly Cys Gly Asn Ser Leu Arg Thr
515 520 525
Ala Leu Ile Asn Ser Thr Gly Glu Glu Val Ala Met Arg Lys Leu Val
530 535 540
Arg Ser Val Thr Val Val Glu Asp Asp Glu Asp Glu Asp Gly Asp Asp
545 550 555 560
Leu Leu His His His His Gly Ser His Cys Ser Ser Ser Gly Asp Pro
565 570 575
Ala Glu Tyr Asn Leu Arg Ser Arg Thr Val Leu Cys Gly Thr Cys Gly
580 585 590
Gln Pro Ala Asp Lys Ala Ser Ala Ser Gly Ser Gly Ala Gln Val Gly
595 600 605
Gly Pro Ile Ser Ser Gly Ser Ser Ala Ser Ser Val Thr Val Thr Arg
610 615 620
Ser Tyr Arg Ser Val Gly Gly Ser Gly Gly Gly Ser Phe Gly Asp Asn
625 630 635 640
Leu Val Thr Arg Ser Tyr Leu Leu Gly Asn Ser Ser Pro Arg Thr Gln
645 650 655
Ser Pro Gln Asn Cys Ser Ile Met
660

Claims (2)

1. application of the nuclear lamina protein A in preparation cutaneum carcinoma Postoperative determination assessment kit, it is characterised in that: with nuclear fabric layer protein White A is as molecular labeling, using source of people nuclear lamina protein A monoclonal antibody or source of people nuclear lamina protein A polyclonal antibody, in conjunction with Immunohistochemical experiment reagent detects relative expression quantity of the nuclear lamina protein A in cutaneous carcinoma tissue.
2. application of the nuclear lamina protein A according to claim 1 in preparation cutaneum carcinoma Postoperative determination assessment kit, Be characterized in that: the source of people nuclear lamina protein A polyclonal antibody is that nuclear lamina protein A shown in SEQ ID NO.1 is exempted from as sequence Epidemic disease rabbit obtains.
CN201910305485.8A 2019-04-16 2019-04-16 Application of the nuclear lamina protein A in preparation cutaneum carcinoma Postoperative determination assessment kit Withdrawn CN110018317A (en)

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Application publication date: 20190716