CN107024587A - A kind of lung cancer early diagnosis marker and diagnostic kit and application - Google Patents
A kind of lung cancer early diagnosis marker and diagnostic kit and application Download PDFInfo
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- CN107024587A CN107024587A CN201710453002.XA CN201710453002A CN107024587A CN 107024587 A CN107024587 A CN 107024587A CN 201710453002 A CN201710453002 A CN 201710453002A CN 107024587 A CN107024587 A CN 107024587A
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- 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
- G01N33/57423—Specifically defined cancers of lung
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Abstract
The present invention relates to a kind of lung cancer early diagnosis marker and diagnostic kit and application, identification marker thing can be early diagnosed as lung cancer by disclosing thromboxane, contrasted by the thrombus cellulose content to experimental group and control group, it was found that content difference of the thromboxane in experimental group and control group is away from very notable, it is suitably applied the antidiastole of lung cancer early stage, with good application value, a kind of new methods for clinical diagnosis is provided for the early diagnosis of lung cancer.
Description
Technical field
Early diagnose and indicate as a kind of lung cancer the present invention relates to technical field of biomedical detection, more particularly to thromboxane
Thing and diagnostic kit and application.
Background technology
Lung cancer has become the global death rate and incidence of disease highest cancer, and epidemiology and results of laboratory are confirmed
85-90% lung cancer is relevant with smoking.Also lacking effective lung cancer early diagnosis marker at present is used for the lung cancer relevant with smoking
Early diagnosis.
Thromboxane is with very strong blood vessel and bronchoconstriction effect, while there is very strong platelet aggregation effect again.Make
For a kind of prostaglandin being metabolized from cell membrane phospholipid, thromboxane relates generally to PUD D, bronchus in lung and roared
Breathe heavily and pulmonary hypertension etc., it has many physiologically actives, including allergy, immune response, angiogenesis and tumour turn
Move etc..
The content of the invention
Present invention aims at a kind of new application there is provided thromboxane, specifically thromboxane is early diagnosed as lung cancer and marked
The application of will thing, and preparing lung cancer early diagnosis kit and application.
The purposes of thromboxane, it is used for the antidiastole mark as lung cancer early stage.
The diagnostic reagent or kit of lung cancer early diagnosis marker as described above are included, the kit contains thrombus
Element is used for the diagnostic reagent for preparing lung cancer Early Identification.
The invention also discloses the application for including lung cancer early diagnosis marker as described above, by test experience group and right
Detection comparative analysis is carried out according to the thrombus cellulose content in the lung tissue or serum of group.
Wherein, detection comparative analysis is carried out to the thrombus cellulose content in lung tissue using liquid phase second order mses method.
Wherein, detection pair is carried out to the thrombus cellulose content of test group and control group using enzyme-linked immunosorbent assay method
Than analysis.
To achieve the above object, the present invention is adopted the technical scheme that：
The purposes of the differential diagnosis kit of the lung cancer, for the diagnostic reagent or kit of the discriminating for preparing lung cancer.
Inventor is all in 4W-24W experiment by the experimental group and the control group of injecting normal saline induced nitrosamine
In the range of mouse pathologic section contrasted, find experimental group pathologic section in thrombus cellulose content it is substantially high
The thrombus cellulose content in the pathologic section of control group；Blood in contrast experiment's group serum in thromboxane and control group serum
Bolt cellulose content, it is found that its significant difference is consistent with thrombus cellulose content significant difference rule in lung tissue, further determine that blood
Application value of the bolt element in lung cancer early diagnosis marker, therefore thromboxane is suitably applied the diagnosis of lung cancer Early Identification.
Beneficial effects of the present invention are as follows：
The present invention is analyzed by thromboxane in the experimental group and the lung tissue of control group and serum to lung cancer, with bright
Aobvious significant difference, can be used as lung cancer early diagnosis marker, can be to be applicable.
Brief description of the drawings
Below in conjunction with the accompanying drawings, it is described in detail by the embodiment to the present invention, technical scheme will be made
And other beneficial effects are apparent.
Experimental period is 4 weeks experimental group mouse pathologic slice maps in Fig. 1（Toluidine blue × 200）；
Experimental period is 4 weeks control group mouse pathologic slice maps in Fig. 2（Toluidine blue × 200）；
Experimental period is 8 weeks experimental group mouse pathologic slice maps in Fig. 3（Toluidine blue × 200）；
Experimental period is 8 weeks control group mouse pathologic slice maps in Fig. 4（Toluidine blue × 200）；
Experimental period is 12 weeks experimental group mouse pathologic slice maps in Fig. 5（Toluidine blue × 200）；
Experimental period is 12 weeks control group mouse pathologic slice maps in Fig. 6（Toluidine blue × 200）；
Experimental period is 16 weeks experimental group mouse pathologic slice maps in Fig. 7（Toluidine blue × 200）；
Experimental period is 16 weeks control group mouse pathologic slice maps in Fig. 8（Toluidine blue × 200）；
Experimental period is 20 weeks experimental group mouse pathologic slice maps in Fig. 9（Toluidine blue × 200）；
Experimental period is 20 weeks control group mouse pathologic slice maps in Figure 10（Toluidine blue × 200）；
Experimental period is 24 weeks experimental group mouse pathologic slice maps in Figure 11（Toluidine blue × 200）；
Experimental period is 24 weeks control group mouse pathologic slice maps in Figure 12（Toluidine blue × 200）；
Figure 13 is Figure 10 enlarged diagram（Toluidine blue × 400）；
Figure 14 is Figure 11 enlarged diagram（Toluidine blue × 400）；
Figure 15 is Figure 12 enlarged diagram（Toluidine blue × 400）；
Figure 16 is thrombus cellulose content average value and mean difference figure in experimental group and control group serum；
Figure 17 is thrombus cellulose content average value and mean difference figure in experimental group and control group lung tissue.
Embodiment
Further to illustrate the technological means and its effect of the invention taken, below in conjunction with being preferable to carry out for the present invention
Example and its accompanying drawing are described in detail.
The purposes of thromboxane, it is used for the antidiastole mark as lung cancer early stage.
The diagnostic reagent or kit of lung cancer early diagnosis marker as described above are included, the kit contains thrombus
Element is used for the diagnostic reagent for preparing lung cancer Early Identification.
The invention also discloses the application for including lung cancer early diagnosis marker as described above, by test experience group and right
Detection comparative analysis is carried out according to the thrombus cellulose content in the lung tissue or serum of group.
Wherein, detection comparative analysis is carried out to the thrombus cellulose content in lung tissue using liquid phase second order mses method.
Wherein, detection pair is carried out to the thrombus cellulose content of test group and control group using enzyme-linked immunosorbent assay method
Than analysis.
Embodiment
Lung cancer mouse pathologic section detection thrombus cellulose content and with the lung tissue thrombus cellulose content of control group
Compare, comprise the following steps that
1st, standby sample：
Inventor selects the SPF level A/J female mices of 60 8 week old, body weight（20±3）G, mouse is purchased from Nanjing University's pattern and moved
Thing research institute, licensing, which is numbered, is：S (XK Soviet Unions) 201000001.220mg nitrosamine accurately is weighed, 11mL physiological saline is dissolved in
In, the 20mgmL-1 NNK aqueous solution is formed, is kept in dark place standby in -20 DEG C of refrigerators.By mouse with 0.1% picric acid dye liquor
It is numbered according to 1-60 numbers and records body weight, is divided into two big groups, every group of 30 groups of mouse according to randomized blocks principle.Pass through
Excel softwares are according to modeling time cycle packet such as table 1, the mouse every of experimental group 30 mouse intraperitoneal injection 0.1mL NNK solution
（20 mg·mL-1）；0.1mL physiological saline is injected intraperitoneally in the mouse every of control group 30 mouse, then according to the length of modeling time
Draw materials in batches.
The mice group table of table 1
| Date group | 4 weeks | 8 weeks | 12 weeks | 16 weeks | 20 weeks | 24 weeks |
| Control group（Only） | 5 | 5 | 5 | 5 | 5 | 5 |
| Blank group（Only） | 5 | 5 | 5 | 5 | 5 | 5 |
2nd, sample
Sampling is divided into serum sample and lung tissue sampling, puts to death 5 mouse according to modeling batch in table 1, by control group
Blood sampling is carried out with the lung tissue and mouse orbit of the mouse of experimental group, lung tissue is placed in equipped with 10% neutral formalin fixer
Vial in preserve, blood sample is placed in 4 DEG C of refrigerators and refrigerates 4h, centrifuging and taking serum is preserved in -20 DEG C of refrigerators.
3rd, sample treatment
3.1 wax stones processed：Transparent, the waxdip bag by experimental group and the lung tissue of control group progress graded ethanol, dehydration of alcohol, dimethylbenzene
Bury and wax stone is made；
3.2 section：Paraffin mass is fixed on slicer, cuts 4 μm of slabs, is placed in the exhibition piece machine equipped with 42 DEG C of water, to be cut
After piece flattening completely, take most intact person on the slide that egg white glycerine was smeared, then put baking box（65℃）Dewax 4~6h；
3.3 dyeing：Dimethylbenzene transparent processing, gradient alcohol dehydration are dried, and distill water process, and haematoxylin is redyed, and circulating water is slightly washed,
Acidic alcohol differentiation immersion liquid processing, washing, running water, which is rinsed, returns indigo plant, and eosin stains distill water process, and gradient alcohol dehydration is done
Dry, neutral gum mounting is made in dimethylbenzene transparent processing.
4th, test：
4.1 micro- sem observations：
The all numbers of obtained experiment are placed under microscope for the neutral gum mounting of 4W, 8W, 12W, 16W, 20W, 24W control group and seen
Examine, such as Fig. 1, Fig. 3, Fig. 5, Fig. 7, Fig. 9, Figure 11, be 4W, 8W, 12W, 16W, 20W, 24W experimental group by all numbers of obtained experiment
Neutral gum mounting is placed in micro- Microscopic observation, such as Fig. 2, Fig. 4, Fig. 6, Fig. 8, Figure 10, Figure 12, finds experimental group compared to control
Group has obvious cancerous lung tissue canceration sign.Wherein Figure 13, Figure 14, Figure 15 are Figure 10, Figure 11, Figure 12 enlarged diagram.
4.2 set up the LC-MS/MS of vertical improvement（Liquid phase second order mses method）Method is examined to the thrombus cellulose content in mouse tissue
Survey, while using Elisa（Enzyme-linked immunosorbent assay method）Detection kit is analyzed, and is inquired into thromboxane and is lured with NNK
The A/J mice lung cancers led occur, the relation of development, are that the diagnosis, prognosis and the research of new drug action target spot of tumour lay the first stone.
Thromboxane content measurement in 4.2 serum
It refer to table 2 and be the blood in the experimental group and control group mice serum that are 4W, 8W, 12W, 16W, 20W, 24W experimental period
The content of bolt element（ng/ml）Contrast table, experimental group and control group that it is 4W, 8W, 12W, 16W, 20W, 24W experimental period that Figure 16, which is,
The content average value and average value disparity map of thromboxane in mice serum, can therefrom be invented, the experiment that all numbers of experiment are 4W
Organize and the thromboxane content difference significance value of control group is 0.73, average value is high by 1.4；The all numbers of experiment are 8W experimental group and right
Thromboxane content difference significance value according to group is 0.08, and average value is high by 5.9；Experimental group and control group that all numbers of experiment are 12W
Thromboxane content difference significance value be 0.14, average value is high by 6.9；The all numbers of experiment are 16W experimental group and the blood of control group
Bolt cellulose content significance of difference value is less than 0.01, and average value is high by 17.8；The all numbers of experiment are 20W experimental group and the thrombus of control group
Cellulose content significance of difference value is less than 0.01, and average value is high by 19.6；The all numbers of experiment are 24W experimental group and the thromboxane of control group
Content difference significance value is less than 0.01, and average value is high by 23.6.
Therefrom it can be found that thromboxane have in the serum early diagnosis of cases of lung cancer it is significant significant, it is adaptable to
For the diagnosis different of lung cancer early stage, a kind of new diagnostic method is provided for lung cancer early diagnosis.
The thrombus cellulose content table of comparisons in the experimental group of table 2 and control group serum
4.3 thromboxane content measurement in lung tissue
Refer to table 3 and be experimental period 4W, 8W, 12W, 16W, 20W, 24W experimental group and control group mice lung tissue in
Content (ng/g) contrast table of thromboxane, experimental group and control that it is 4W, 8W, 12W, 16W, 20W, 24W experimental period that Figure 17, which is,
The content average value and average value disparity map of thromboxane in group mouse lung tissue, can therefrom be invented, and all numbers of experiment are 4W's
Experimental group and the thromboxane content difference significance value of control group are 0.73, and average value is high by 1.4；The experimental group that all numbers of experiment are 8W
Thromboxane content difference significance value with control group is 0.08, and average value is high by 5.9；The all numbers of experiment are 12W experimental group and right
Thromboxane content difference significance value according to group is 0.14, and average value is high by 6.9；Experimental group and control group that all numbers of experiment are 16W
Thromboxane content difference significance value be less than 0.01, average value is high by 17.8；Experimental group and control group that all numbers of experiment are 20W
Thromboxane content difference significance value is less than 0.01, and average value is high by 19.6；The all numbers of experiment are 24W experimental group and the blood of control group
Bolt cellulose content significance of difference value is less than 0.01, and average value is high by 23.6.
Therefrom it can be found that thromboxane have in the lung tissue early diagnosis of cases of lung cancer it is significant significant, with blood
Thrombus cellulose content experimental group is consistent with the difference rule of control group in clear, further proves that thromboxane is applied to be used for lung cancer early stage
Diagnosis different, for lung cancer early diagnosis provide a kind of new diagnostic method.
The thrombus cellulose content table of comparisons in the experimental group of table 3 and the lung tissue of control group
Mouse of the content of thromboxane in the lung tissue and serum of the mouse of the experimental group obtained by test apparently higher than control group
The content of the thromboxane of lung tissue, with obvious significant difference, can be used, this hair as lung cancer early diagnosis marker
It is bright to provide a kind of new diagnostic method for lung cancer early diagnosis.
It is described above, for the person of ordinary skill of the art, can be with technique according to the invention scheme and technology
Other various corresponding changes and deformation are made in design, and all these changes and deformation should all belong to the claims in the present invention
Protection domain.
Claims (5)
1. the purposes of thromboxane, it is characterised in that for the antidiastole mark as lung cancer early stage.
2. include the diagnostic reagent or kit of lung cancer early diagnosis marker as claimed in claim 1, it is characterised in that institute
State kit and contain the diagnostic reagent that thromboxane is used to prepare lung cancer Early Identification.
3. include the application of lung cancer early diagnosis marker as claimed in claim 1, it is characterised in that by test experience group and
Thrombus cellulose content in the lung tissue or serum of control group carries out detection comparative analysis.
4. application according to claim 3, it is characterised in that using liquid phase second order mses method to the thrombus in lung tissue
Cellulose content carries out detection comparative analysis.
5. application according to claim 3, it is characterised in that using enzyme-linked immunosorbent assay method to test group and
The thrombus cellulose content of control group carries out detection comparative analysis.
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| CN201710453002.XA CN107024587A (en) | 2017-06-15 | 2017-06-15 | A kind of lung cancer early diagnosis marker and diagnostic kit and application |
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108998530A (en) * | 2018-08-31 | 2018-12-14 | 昆明医科大学第附属医院 | Lung cancer raises long-chain non-coding RNA marker and its application |
| CN108998531A (en) * | 2018-08-31 | 2018-12-14 | 昆明医科大学第附属医院 | Lung cancer lowers long-chain non-coding RNA marker and its application |
| CN110699457A (en) * | 2019-10-30 | 2020-01-17 | 深圳瑞科生物科技有限公司 | Primer group and kit for detecting lung cancer |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102520156A (en) * | 2011-12-15 | 2012-06-27 | 苏州苏大赛尔免疫生物技术有限公司 | Enzyme linked immunoassay kit of thromboxane B2 (TXB2) |
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2017
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102520156A (en) * | 2011-12-15 | 2012-06-27 | 苏州苏大赛尔免疫生物技术有限公司 | Enzyme linked immunoassay kit of thromboxane B2 (TXB2) |
Non-Patent Citations (2)
| Title |
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| 孙京栋等: "血栓素A2、前列环素与Lewis肺癌生长及转移关系的实验研究", 《中国肿瘤临床》 * |
| 张永生等: "《检验项目与临床应用》", 31 December 2014 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108998530A (en) * | 2018-08-31 | 2018-12-14 | 昆明医科大学第附属医院 | Lung cancer raises long-chain non-coding RNA marker and its application |
| CN108998531A (en) * | 2018-08-31 | 2018-12-14 | 昆明医科大学第附属医院 | Lung cancer lowers long-chain non-coding RNA marker and its application |
| CN108998531B (en) * | 2018-08-31 | 2021-10-01 | 昆明医科大学第一附属医院 | Long-chain non-coding RNA marker for lung cancer down-regulation and application thereof |
| CN110699457A (en) * | 2019-10-30 | 2020-01-17 | 深圳瑞科生物科技有限公司 | Primer group and kit for detecting lung cancer |
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