CN102662060A - Application of Matriptase - Google Patents
Application of Matriptase Download PDFInfo
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- CN102662060A CN102662060A CN2012101140128A CN201210114012A CN102662060A CN 102662060 A CN102662060 A CN 102662060A CN 2012101140128 A CN2012101140128 A CN 2012101140128A CN 201210114012 A CN201210114012 A CN 201210114012A CN 102662060 A CN102662060 A CN 102662060A
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- matriptase
- chronic lymphocytic
- diagnosis
- leukemia
- lymphocytic leukemia
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Abstract
The invention discloses an application of Matriptase. The Matriptase comprises a short intracellular segment, a transmembrane region, an SEA (sea urchin sperm protein, enteropeptidase, agrin domain) region, two CUB (Cls/Clr, urchin embryonic growth factor and bone morphogenetic protein-1) regions, four low density lipoprotein acceptor domains and a C-terminal trypsin-like protease region. The Matriptase is applied to diagnosis and treatment of chronic lymphocyte leukemia. By an experiment, bone marrows and peripheral blood specimens of normal people and leukemia sufferers are collected, and the detection result shows as follows: Matriptase mRNA and protein are highly expressed in the chronic lymphocyte leukemia sufferers, the expression level of Matriptase of the surface of lymphocyte can serve as a molecular marker for diagnosis of chronic lymphocyte leukemia, and the Matriptase can serve as a novel target for diagnosis and treatment of chronic lymphocyte leukemia.
Description
Technical field
The present invention relates to biological technical field, specifically, is the effect of Matriptase in chronic lymphocytic leukemia medical diagnosis on disease, treatment, therapeutic evaluation and prognosis.
Background technology
Leukaemia is a kind of pernicious neoplastic hematologic disorder, and its incidence of disease increases year by year, and wherein chronic lymphocytic leukemia is that B is a lymphocyte opposing apoptosis, and clone's property is accumulated in lymphoid tissue, marrow and peripheral blood, causes the depleted neoplastic hematologic disorder of normal hematopoiesis eventually.Chronic lymphocytic leukemia is about at western countries' incidence of disease that all are leukemic 30%, and China and Asian countries's incidence of disease are lower than 10%, and increase trend is arranged in recent years.The characteristics of chronic lymphocytic leukemia cell are that viability is strong, and apoptosis is suppressed, and its pathogenesis it be unclear that.Therefore, further probe into the new target drone of this medical diagnosis on disease and treatment, have great importance and clinical value.
Matriptase has another name called MT-SP1, TADG-15, and epithin and ST-14 belong to II type transmembrane serine protease family, in external breast cancer cell nutrient culture media, are identified out to have the activity of gelatin hydrolysate in 1993.The encoding gene ST14 of Matriptase is positioned at human chromosome 11q24-25,855 the amino acid whose polypeptide of encoding.Matriptase is expressed in embryo and adult's the epithelial tissue usually.In addition, in immunocyte such as mast cell, monocyte and macrophage, also can detect the expression of Matriptase.
Confirmed that at present Matriptase has vital role at aspects such as epithelium differentiation, skin function, immune systems, and closely related with multiple entity tumor.From breast cancer cell the earliest, come to light so far,, in kidney, liver, lungs and ovarian neoplasm, the SCCHN, all can detect the overexpression of Matriptase at prostate, colon; In the part tumour, the Matriptase high level expression is bad relevant with clinical prognosis.People such as List have set up skin and have crossed the transgenic mice of expressing Matriptase, and they find that Matriptase expresses to increase and can cause the spontaneous squamous cell carcinoma of mouse and the susceptibility that chemicals brings out cutaneum carcinoma is strengthened.In addition, with cross express Matriptase stomach cancer cell through lumbar injection in nude mouse, tumour cell increases nearly 3 times to the ability of lymphatic metastasis diffusion.The external invasive ability that can also significantly strengthen colon cancer cell is expressed in crossing of Matriptase.These researchs show that increasing of Matriptase expression can promote tumour to form and transfer.
Discover that recently Matriptase has trace expression at peripheral blood mononuclear and macrophage, but its biological function in blood is still not clear.Our early-stage Study result finds that Matriptase mRNA and albumen are expressed all unusually in the patients with chronic lymphocytic sample increase; Show that the high expressed of Matriptase in chronic lymphocytic leukemia has its specificity, prompting Matriptase possibly play a significant role in chronic lymphocytic leukemia course of disease progress.Its specific expressed changing may become the new molecular marker of chronic lymphocytic leukemia, becomes new chronic lymphocytic leukemia diagnosis and treatment target spot.
Summary of the invention
The present invention is directed to present Matriptase expresses at the patients with chronic lymphocytic lymphocytic cell surface; The invention discloses the application of Matriptase in chronic lymphocytic leukemia diagnosis and treatment; Be used for chronic lymphocytic leukemia medical diagnosis on disease index, and as chronic lymphocytic leukemia treatment novel targets.
Matriptase has another name called MT-SP1, TADG-15, and epithin and ST-14 belong to II type transmembrane serine protease family, in external breast cancer cell nutrient culture media, are identified out to have the activity of gelatin hydrolysate in 1993.The encoding gene ST14 of Matriptase is positioned at human chromosome 11q24-25,855 the amino acid whose polypeptide of encoding.
The application of Matriptase, said Matriptase comprises born of the same parents' inner segment of a weak point, strides the film district for one; A SEA district (sea urchin sperm protein; Enteropeptidase, agrin domain), two CUB district (Cls/Clr; Urchin embryonic growth factor and bone morphogenetic protein-1); Four LDL receptors (LDLRA) domain, and the trypsin-like proteinase zone of C end, said Matriptase is applied in chronic lymphocytic leukemia diagnosis and the treatment.
Further, said Matriptase is used for the chronic lymphocytic leukemia diagnosis index.
Further, said Matriptase newly harrows a little as the treatment of chronic lymphocytic leukemia.
Compared with prior art, the present invention has the following advantages:
Through experiment; Collect normal person and leukaemic's marrow and peripheral blood sample; The testing result prompting: Matriptase mRNA and albumen high expressed are in patients with chronic lymphocytic; The expression of prompting lymphocytic cell surface Matriptase can be used as the molecular marker of chronic lymphocytic leukemia diagnosis, is used for the novel targets of chronic lymphocytic leukemia diagnosis and treatment.
Above-mentioned explanation only is the general introduction of technical scheme of the present invention, understands technological means of the present invention in order can more to know, and can implement according to the content of instructions, below with preferred embodiment of the present invention and conjunction with figs. specify as after.
Description of drawings
Accompanying drawing described herein is used to provide further understanding of the present invention, constitutes the application's a part, and illustrative examples of the present invention and explanation thereof are used to explain the present invention, do not constitute improper qualification of the present invention.In the accompanying drawings:
Fig. 1 is the schematic arrangement of Matriptase;
Fig. 2 detects the result of Matriptase for RT-PCR;
Fig. 3 detects the result of Matriptase for Western blot;
Fig. 4 detects the gray-scale statistical result of Matriptase for Western blot;
Fig. 5 is the result that flow cytometry and immunofluorescence dyeing detect Matriptase.
Embodiment
Below with reference to accompanying drawing and combine embodiment, specify the present invention.
The application of Matriptase, said Matriptase is applied in chronic lymphocytic leukemia diagnosis and the treatment.Referring to shown in Figure 1, said Matriptase comprises born of the same parents' inner segment of a weak point, strides the film district for one; A SEA district (sea urchin sperm protein; Enteropeptidase, agrin domain), two CUB district (Cls/Clr; Urchin embryonic growth factor and bone morphogenetic protein-1); Four LDL receptors (LDLRA) domain, and the trypsin-like proteinase zone of C end, said Matriptase is applied in chronic lymphocytic leukemia diagnosis and the treatment.
Further, said Matriptase is used for the chronic lymphocytic leukemia diagnosis index.
Further, said Matriptase newly harrows a little as the treatment of chronic lymphocytic leukemia.
The practical implementation process:
One. gather leukaemic's marrow and peripheral blood
Gather 143 routine leukaemics' sample of bone marrow, comprise 74 routine chronic lymphocytic leukemias, 20 routine ALLs, 28 routine acute myeloblastic leukemias and 21 routine chronic myelocytic leukemias.
Two, detect said Matriptase mRNA expression
Extract total RNA of all leukaemic's myeloplasts; Adopt RT-PCR and real-time quantitative RT-PCR (Power SYBR Green PCR Master Mix; Applied Biosystems) expression of the said Matriptase mRNA of detection; Find that through detecting (72/74,97.3%) said Matriptase mRNA expresses increase in most chronic lymphocytic leukemia cases, in other types of leukemia encountered (ALL, acute myeloblastic leukemia and chronic myelocytic leukemia); Basically not observing said Matriptase mRNA expresses; Referring to shown in Figure 2, be part RT-PCR testing result, wherein each 10 example of chronic lymphocytic leukemia and ALL; Each 6 example of chronic myelocytic leukemia and acute myeloblastic leukemia, ctrl is not for adding the negative control of cDNA template.
Three, Western blot, flow cytometry and immunofluorescence technique detect said Matriptase protein expression situation:
1, extract clone and leukaemic's myeloplast total protein, BCA (bicinchoninic acid) method is measured protein concentration, and Western blot detects the expression of said Matriptase albumen.Referring to Fig. 3 and shown in Figure 4; Western blot result shows; The expression of Matriptase albumen described in patients with chronic lymphocytic (n=18) bone marrow cell is obviously increased, and does not detect the expression of said Matriptase albumen in ALL (n=12), acute myeloblastic leukemia (n=11) and chronic myelocytic leukemia (n=3) the patient bone marrow cell.NPBC is the contrast of normal person's marrow among the figure, and n is a sample example number.Wherein A is the Western blot result of part sample, and B is the gray-scale statistical result, * * p < 0.01 (other group of chronic lymphocytic leukemia vs.).
2, extract clone and leukaemic's myeloplast total protein, BCA (bicinchoninic acid) method is measured protein concentration, and flow cytometry and immunofluorescence dyeing detect location and the expression of said Matriptase albumen at surface of cell membrane.Referring to shown in Figure 5; In the expression that said Matriptase albumen can be clearly seen on the lymphoma cell line Namalwa cell and the patients with chronic lymphocytic bone marrow cell surface in B cell source, still then do not observe the expression of said Matriptase albumen on the surface of normal person's PBC (NPBC).To be uniformly coated on the microslide with the cell smear machine with the above-mentioned cell behind the fluorescence antibody mark, with DAPI-Fluoromount-G (SouthernBiotech) mountant mounting, Laser Scanning Confocal Microscope (Leica TCS-SP2) is observed down.The result is: the expression of said Matriptase albumen is almost feminine gender (0.5%) on the NPBC surface; And in the whole basically positive expression of Matriptase albumen described in Namalwa cell and the patients with chronic lymphocytic bone marrow cell, positive rate is respectively 99% and 93%.This result confirms that said Matriptase is positioned chronic lymphocytic leukemia sample surface of cell membrane; Thereby point out processes such as infiltration that said Matriptase possibly participate in the chronic lymphocytic leukemia cell and surface signal conduction, the flow cytometry result also points out Matriptase to be used for the diagnosing chronic lymphocytic leukemia as the cell surface marker thing.Bar=5 μ m among the figure.
The above is merely the preferred embodiments of the present invention, is not limited to the present invention, and for a person skilled in the art, the present invention can have various changes and variation.All within spirit of the present invention and principle, any modification of being done, be equal to replacement, improvement etc., all should be included within protection scope of the present invention.
< 110>University Of Suzhou
< 120>application of Matriptase
<160> 2
<210> 1
<211> 3319
<212> DNA
< 213>artificial sequence
<400> 1
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< 213>artificial sequence
<400> 2
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271?RGS?DLV?TVY?NTL?SPM?EPH?ALV?QLC?GTY?PPS?YNL?TFH?SSQ?NVL?LIT
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496?CKN?KFC?KPL?FWV?CDS?VND?CGD?NSD?EQG?CSC?PAQ?TFR?CSN?GKC?LSK
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586?ECD?GKE?DCS?DGS?DEK?DCD?CGL?RSF?TRQ?ARV?VGG?TDA?DEG?EWP?WQV
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Claims (3)
1.Matriptase application; Said Matriptase comprises born of the same parents' inner segment of a weak point, strides the film district for one, a SEA district; Two CUB districts; Four LDL receptor domains, and the trypsin-like proteinase zone of C end, it is characterized in that: said Matriptase is applied in chronic lymphocytic leukemia diagnosis and the treatment.
2. the application of Matriptase according to claim 1 is characterized in that: said Matriptase is used for the chronic lymphocytic leukemia diagnosis index.
3. the application of Matriptase according to claim 1 is characterized in that: said Matriptase newly harrows a little as the treatment of chronic lymphocytic leukemia.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN2012101140128A CN102662060A (en) | 2012-04-18 | 2012-04-18 | Application of Matriptase |
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| CN2012101140128A CN102662060A (en) | 2012-04-18 | 2012-04-18 | Application of Matriptase |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110391016A (en) * | 2019-07-10 | 2019-10-29 | 浙江大学 | A kind of analysis method of Dynamic constrasted enhancement magnetic resonance image |
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2012
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110391016A (en) * | 2019-07-10 | 2019-10-29 | 浙江大学 | A kind of analysis method of Dynamic constrasted enhancement magnetic resonance image |
| CN110391016B (en) * | 2019-07-10 | 2021-11-09 | 浙江大学 | Analysis method for dynamic contrast enhanced magnetic resonance image |
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Application publication date: 20120912 |