CN106950384A - Rectal neoplasm mark and its application, kit - Google Patents
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Abstract
The invention belongs to biomedicine technical field, specifically related to a kind of rectal neoplasm mark, the rectal neoplasm mark is Beclin1 albumen, and it is a kind of for detecting that the expression of the Beclin1 albumen and/or the reagent of phosphorylation level are being prepared for detection and/or Diagnosis of Rectal tumour or for the application in the treatment to rectal neoplasm patient and/or the kit of prognostic evaluation, a kind of detection and/or Diagnosis of Rectal tumour or the treatment to intestinal tumor patient and/or the kit of prognostic evaluation, including detecting the expression of Beclin1 albumen and/or the reagent of phosphorylation level.The mark aggregate level and phosphorylation level by joint assessment, can as rectal neoplasm early detection and/or the reference of diagnosis, and rectal neoplasm patient Medication monitor, the reference of drug resistance prediction.
Description
Technical field
The invention belongs to biomedicine technical field, and in particular to a kind of rectal neoplasm mark and its application, kit.
Background technology
Cell autophagy (Autophagy) is one kind " reversal " phenomenon of multicellular organism, and cell is by wrapping up endochylema content
Thing is conveyed to the degraded of lyase system to recycle the need that catabolite supplies normal activity, and the phenomenon has evolution conservative
Property.Autophagy is the important defence of body and protection mechanism, and the complicated and angle of multiple-effect is play in many physiology and pathologic process
Color.When cells survival is on the hazard, such as anoxic or nutritional deficiency, autophagy can be survived non-emergent albumen by lysosomal degradation
With organelle etc., metabolic nutrient raw material is recycled to promote cell survival；Autophagy can also degrade and eliminate toxic
The outmoded albumen and protein masses of tendency, form protection mechanism to resist nerve retrograde affection；In addition, autophagy can also be gulped down
Bacterium and the virus of invasion are bitten, is locked it in autophagosome, lysosome is finally transported to and eliminates to play the work of immune defense
With.The disorder of autophagocytosis can cause many pathological reactions, and tumour is one of result of its dysfunction.
Autophagy needs the synergy of multiple signal paths and multienzyme complex, Beclin1 (Bcl2-binding myosin-
Like protein 1) be autophagy molecular mechanism nucleus, be the skelemin in Beclin1-VPS34 complexs.
VPS34 is mammal type III phosphatidylinositol3 3 kinase (PI3K), the phosphatide of its specifically three, Phosphation inose ring
Sour inositol is simultaneously translated into the phosphate of phosphatidic acid inositol 3 (PI3Ps).The activation of VPS34 cytoplasme lcinases activity is that autophagy starts
Committed step in mechanism because the mark member of autophagy process-be used for wrapping up double film autophagosomes of endochylema content-formation
Need the PI3Ps largely produced by VPS34.Beclin1 is combined closely with VPS34, and interaction is provided for autophagy regulatory factor
Platform, therefore it autophagy regulation and control in play uniqueness role.As far as is known, multiple autophagy regulatory factors by with
Beclin1 interacts and forms the unique Beclin1-VPS34 secondary composites body of function to adjust VPS34 activity, so that
Play the adjustment effect each to autophagy process.The regulation of Beclin1-VPS34 complexs activity with autophagy except by regulating and controlling
The combination of the factor beyond realizing, can also be realized by Beclin1 phosphorylations, so as to change autophagy intensity.Beclin1 can be
Some positions are phosphorylated, so as to influence the formation of Beclin1-VPS34 complexs, it is then sharp and exactly for intracellular
Many influence factors produce response.
Rectal neoplasm is from dentate line between sigmoid colon intersection as formed by occurring canceration for rectal tissue cell
Tumour, its position gos deep into pelvic cavity, and recurrence after operation rate is high.Its origin cause of formation is unknown, it is now recognized that high risk factor be related to nutrition not
The too high and food fiber insufficiency of intake of equilibrium, i.e. animal tallow and protein intake.The incidence of disease of rectal neoplasm increases year by year, only
It is following or lung cancer and stomach cancer can be exceeded inferior to lung cancer and stomach cancer, leap to the first, therefore the diagnosis and treatment of rectal neoplasm are current
The particularly important research topic in the world.Specific biomarkers involved by the current diagnostic process of current rectal neoplasm have cancer
Two kinds of embryonal antigen (CEA) and sugar chain antigens 19-9 (CA19-9), the former is used to diagnose and detect, the latter is used for prognosis evaluation, two
Person must use in conjunction, but fault rate still exists, it is necessary to more detection means are to prove, such as iconography and endoscopy, and
These detection methods have respective shortcoming, such as take, have traumatic, expensive, and body, spirit plus warp are constituted to patient
Ji etc. is multiple to be shared.It is particularly noteworthy that the middle hypomere tumour in rectal neoplasm is because close to anal sphincter, causing art
Anus preserving turns into a problem afterwards, and this also make it that rectal neoplasm turns into a kind of disease for maximum of being disputed on operation method, clinical
The strategy that doctor takes is typically " first save one's life, then protect anus ".But because artificial anus is formed by stomach wall colostomy, Huan Zhebi
, because of psychology and social cause, it must thus bring great mental suffering permanently in stomach wall position defecation, have a strong impact on patient
Quality of life and life dignity so that many patients would rather select death to be also reluctant to receive stomach wall artificial anus.
The difficult point of preventing and treating of rectal neoplasm includes：1) early diagnose, due to the individuality of conditions of patients, for early screening
The selection of biomarker is rather difficult so that most patients have been middle and advanced stages when finding；2) anus is protected, can operative treatment keep
Anus, the principal concern as patient；3) drug resistance, chemotherapeutics is due to molecular target poor specificity so that drug resistance and
Drug side-effect is obvious.Therefore personalized target is found very crucial for the preventing and treating of rectal neoplasm.
The content of the invention
It is an object of the invention to overcome the above-mentioned deficiency of prior art there is provided a kind of rectal neoplasm mark and its answer
With, kit, it is intended to solve the technology that specific biomarkers are limited and rectal neoplasm preventing and treating is difficult of existing rectal neoplasm
Problem.
For achieving the above object, the technical solution adopted by the present invention is as follows：
On the one hand, the present invention provides a kind of rectal neoplasm mark, and the rectal neoplasm mark is Beclin1 albumen.
On the other hand, the present invention provides a kind of expression and/or phosphorylation water for being used to detect above-mentioned Beclin1 albumen
Flat reagent is being prepared for detection and/or Diagnosis of Rectal tumour or commented for treatment and/or prognosis to rectal neoplasm patient
Application in the kit of valency.
Finally, the present invention also provides a kind of detect and/or Diagnosis of Rectal tumour or treatment to intestinal tumor patient and/or pre-
The kit evaluated afterwards, the kit includes detecting the examination of the expression and/or phosphorylation level of above-mentioned Beclin1 albumen
Agent.
The rectal neoplasm mark that the present invention is provided be Beclin1 albumen, the expression of the Beclin1 albumen and/or
Phosphorylation level has correlation with rectal neoplasm.We are from cell autophagy angle, with the skelemin in the path
Beclin1 is research object, and the Beclin1 levels in rectal neoplasm clinical sample and phosphorylation level are characterized first.
Beclin1 state can be divided into autophagy promoted type and autophagy suppressive, and the state of autophagy can be divided into the beneficial type of tumour cell
Type is harmful to tumour cell.When tumour is in different (formation and development, shift and infiltrate, and chemicotherapy) by stages, at autophagy
In different states, therefore it can be adjusted by monitoring Beclin1 state (albumen aggregate level and phosphorylation modification level)
The therapeutic scheme of whole tumor patient, such as autophagy promotion-beneficial type of tumour cell (being in Tumor Growth) and autophagy
Suppression-tumour cell is harmful to type and (is in effective therapeutic process) patient, it should select autophagy suppressive chemotherapeutics；For certainly
Bite promotion-tumour cell and be harmful to type (being in effective therapeutic process) and autophagy suppression-beneficial type of tumour cell (in tumour life
In growth process) patient, it should select autophagy promoted type chemotherapeutics.Therefore, Beclin1 state may be used as rectal neoplasm
Early diagnosis and therapeutic effect tracking and prognostic evaluation provide reference.
The present invention is provided in a kind of examination for being used to detect the expression and/or phosphorylation level of above-mentioned Beclin1 albumen
Agent is being prepared for detection and/or Diagnosis of Rectal tumour or the treatment to rectal neoplasm patient and/or the kit of prognostic evaluation
In application, the application be embodied on product be it is a kind of detect and/or Diagnosis of Rectal tumour or treatment to intestinal tumor patient and/
Or the kit of prognostic evaluation.By collecting clinical rectal neoplasm tissue samples, research sufferer personalization expression Beclin1 feelings
Condition, between the individuation and tumour individuation of Beclin1 phosphorylations set up be available for contacting for clinical reference, be clinical diagnosis and
Treatment provides help.The kit can be to the Beclin1 protein levels and Beclin1 phosphorylations in rectal neoplasm clinical sample
The detection of two levels of level, investigation Beclin1 expression and the personalized relation between rectal neoplasm sign personalization of modification,
By joint assessment, it can be supervised as rectal neoplasm early detection and/or the reference of diagnosis, and the medication of rectal neoplasm patient
Survey, the reference of drug resistance prediction.
Brief description of the drawings
Fig. 1 determines external protein B eclin1_WT, Beclin1_ to utilize constant temperature measuring ball method in the embodiment of the present invention 1
229E233E and Atg14L interaction result datagrams；
Fig. 2 is the results of immunoblot analysis figure of Beclin1 phosphorylation condition tests in the embodiment of the present invention 2；
Fig. 3 is clinical rectal neoplasm LC3-I/II and Beclin1/Beclin1 Expression of phosphorylated water in the embodiment of the present invention 3
Flat immunoblotting analysis analysis result figure；
Fig. 4 is that different tissues position Beclin1 expressions are exempted from rectal neoplasm clinical sample in the embodiment of the present invention 4
Epidemic disease histochemical analysis result figure；
Fig. 5 is LC3-I/II and p62 tables in the Hela cells in the embodiment of the present invention 5 after anti-rectal neoplasm drug-treated
Up to horizontal immunoblotting analysis analysis result figure.
Embodiment
In order that technical problems, technical solutions and advantageous effects to be solved by the present invention are more clearly understood, below in conjunction with
Embodiment, the present invention will be described in further detail.It should be appreciated that specific embodiment described herein is only to explain
The present invention, is not intended to limit the present invention.
On the one hand, the embodiments of the invention provide a kind of rectal neoplasm mark, the rectal neoplasm mark is Beclin1
Albumen.Expression and/or phosphorylation level and the rectal neoplasm of the Beclin1 albumen have correlation.Pass through research, autophagy
The personalization of state and the personalized one-to-one corresponding of rectal neoplasm, propose Beclin1 expression and modificationization state, can be from molecular water
The flat upper personal feature for judging rectal neoplasm patient, theoretical foundation is provided for rectal neoplasm personalized treatment.
In one embodiment of the invention, Beclin1 phosphorylations and autophagy activation factor are simulated with Beclin1 mutains
Adhesion between Atg14L.Complete Beclin1 mono- has three functional areas (BH3 areas, CCD areas, ECD areas), and the present invention is real
Example is applied with the sequence (amino acid that numbering is 102-268 in people source Beclin1 partial amino-acid series, i.e. BH3 areas and CCD areas
Sequence) as wild type Beclin1 albumen (abbreviation Beclin1_WT), by two junket ammonia in wild type Beclin1 albumen
Acid (amino acid number difference 229 and 233) simultaneous mutation is that (abbreviation Beclin1_229E233E, E are glutamic acid formation mutant
Glutamic acid, Glutamic acid, trigram is abbreviated as Glu, and one-letter abbreviations are E, and glutamic acid acidic amino acid can be from electric charge
Atom microenvironment after upper simulation phosphorylation), i.e., the mutation body length is initial amino acid numbering 102, terminates amino acid number
268,167 amino acid, is re-introduced into 4 amino acid (4 amino acid " GPGS " most started) of connection expression vector altogether,
Therefore 171 amino acid constitute the Beclin1 mutains of the present invention, the amino acid sequence of the Beclin1 mutains altogether
For SEQ ID NO:1：GPGSEASDGGTMENLSRRLKVTGDLFDIMSGQTDVDHPLCEECTDTLLDQLDTQLNVTENECQ
NYKRCLEILEQMNEDDSEQLQMELKELALEEERLIQELEDVEKNRKIVAENLEKVQAEAERLDQEEAQEQREESEFK
RQQLELDDELKSVENQMRYAQTQLDKLKKTN。
Because the startup of autophagy needs to undergo three steps：The first step, Beclin1_WT-Beclin1_WT homodimer solutions
From being changed into Belcin1_WT monomers；Second step, Beclin1_WT monomers and Atg14L formation Beclin1_WT-Atg14L are heterologous
Dimer；3rd step, Beclin1_WT-Atg14L heterodimers raise corresponding activation factor as platform is combined, and start certainly
Bite.Based on the molecular mechanism of this protein-interacting, in the Beclin1_WT-Beclin1_WT homodimers three parsed
Tie up on the basis of structure, by gene mutation means, devise people source Beclin1_229E233E mutation, it is intended to simulate
Beclin1229, the tyrosine phosphorylation in 233 two sites.As a result show, 229, the tyrosine in 233 two sites is phosphorylated
Afterwards, the interaction between Beclin1_WT and Atg14L will be lowered, further implies that it produces autophagy inhibitory action.
On the other hand, the embodiment of the present invention provides a kind of expression and/or phosphorus for being used to detect above-mentioned Beclin1 albumen
The reagent of acidifying level prepare for detect and/or Diagnosis of Rectal tumour or for the treatment to rectal neoplasm patient and/or
Application in the kit of prognostic evaluation.The embodiment of the present invention is made with Beclin1 albumen aggregate level and Beclin1 phosphorylation levels
Rectal neoplasm is applied to for Testing index to early diagnose with studying, and is detected and/or Diagnosis of Rectal tumour or swollen to rectum with preparing
The treatment of knurl patient and/or the kit of prognostic evaluation.
Preferably, the phosphorylation level of Beclin1 albumen is 229 sites of Beclin1 albumen and the tyrosine in 233 sites
Phosphorylation level.In an embodiment of the present invention, to the condition test of Beclin1 phosphorylations, 229,233 the two sites are shown
Or be EGFR main substrate site, it was demonstrated that it serves as the important role for causing function to change by structural modification.
Preferably, the reagent of the expression of detection Beclin1 albumen includes anti-Beclin1 protein antibodies or fluorescence probe
The Beclin1 protein antibodies of modification, the reagent that the phosphorylation level of Beclin1 albumen is stated in detection is anti-including anti phosphotyrosine
Body.In an embodiment of the present invention, SABC is carried out to clinical rectal cancer tissue and cancer beside organism with anti-Beclin1 antibody
Experimental analysis；In an alternative embodiment of the invention, it is swollen to clinical rectum with anti-Beclin1 antibody and anti phosphotyrosine antibody
The immunoblotting analysis analysis of knurl Beclin1/Beclin1 Expression of phosphorylated levels.
Preferably, detect that the expression of described Beclin1 albumen and/or the method for phosphorylation level are immunology side
Method.In embodiments of the present invention, immunological method includes Western blot, Immunohistochemical Method and co-immunoprecipitation method.
Preferably, to rectal neoplasm patient treatment and/or the kit of prognostic evaluation can join with anti-rectal neoplasm medicine
Share medicine.Anti- rectal neoplasm medicine is included in capecitabine, Irinotecan, oxaliplatin, fluorouracil and Calciumlevofolinate extremely
Few one kind.In an embodiment of the present invention, by the way that to clinical anti-rectal neoplasm medicine autophagy Capability Categories, " oxaliplatin+fluorine is urinated
Pyrimidine+Calciumlevofolinate " combination has autophagy facilitation；And capecitabine and Irinotecan do not have autophagy facilitation, the reagent
Box by the expression and/or phosphorylation level of Beclin1 albumen, obtain relevant information as rectal neoplasm Medication monitor,
The reference of drug resistance prediction.
Finally, the embodiment of the present invention provide it is a kind of detect and/or Diagnosis of Rectal tumour or treatment to intestinal tumor patient and/
Or the kit of prognostic evaluation, the examination of expression and/or phosphorylation level of the kit including above-mentioned Beclin1 albumen
Agent.Specifically, the phosphorylation level of Beclin1 albumen refers to 229 sites of Beclin1 albumen and the tyrosine phosphorus in 233 sites
Acidifying level, the two sites or the main substrate site for EGFR.Specifically, the expression of Beclin1 albumen is detected
Reagent includes the Beclin1 protein antibodies that anti-Beclin1 protein antibodies or fluorescence probe are modified, and detects the phosphorus of Beclin1 albumen
The reagent of acidifying level includes anti phosphotyrosine antibody.
It is of the invention successively to carry out test of many times, now lift A partial experiment result and carried out further in detail as reference pair invention
Thin description, is described in detail with reference to specific embodiment.
The effect simulation of the Beclin1 phosphorylations of embodiment 1
Constant temperature measuring ball method determines external protein B eclin1_WT, Beclin1_229E233E and autophagy regulatory factor
Atg14L is interacted, and the knot between Beclin1 phosphorylations and autophagy activation factor Atg14L is simulated with Beclin1 mutains
With joint efforts.Detailed process is as follows：
The solvable table of people source Beclin1_WT, Beclin1_229E233E, Atg14L albumen is first realized in Escherichia coli
Reach, and isolate and purify and obtain enough destination proteins；Beclin1_WT, Beclin1_229E233E and Atg14L be dialyzed to
Buffer solution (contains 50mM Tris, 150mM NaCl；PH 8.0) in；Isothermal calorimetric titration instrument titration syringe respectively on
The μ l of sample 40 Beclin1_WT (concentration 12mg/ml) and 40 μ l Beclin1_229E233E (concentration 12mg/ml), and in isothermal
The μ l of sample cell loading 220 of calorimetric titration instrument Atg14L (the μ g/ml of concentration 500), utilizes isothermal calorimetric titration instrument iTC200
(MicroCal) respectively between measure autophagy skelemin Beclin1_WT and Atg14L, Beclin1_229E233E and Atg14L
Interaction caused by thermal transition；Determining every time has 10-40 sample introduction, is balanced 180 seconds per between sample introduction twice, and it is counted
Gather and analyze according to Origin 7.0.
By determining binding constant and carrying out quantitative scoring calculation, as a result as shown in Figure 1：From figure knowable to data, than wild
(Figure 1A is that constant temperature calorimetric titration determines Beclin1_WT and Atg14L phases for interaction between type Beclin1_WT and Atg14L
Interaction), (Figure 1B is determined the interaction between saltant type Beclin1_229E233E and Atg14L for constant temperature calorimetric titration
Beclin1_229E233E and Atg14L interacts) weaken, after hint 229, the tyrosine in 233 two sites are phosphorylated,
The interaction between Beclin1_WT and Atg14L will be lowered, further illustrate that it produces autophagy inhibitory action.
The condition test of the Beclin1 phosphorylations of embodiment 2
(229,233 tyrosine sites are directed to Beclin1 wild types (Beclin1_WT) and saltant type using Escherichia coli
It is mutated) heterogenous expression is carried out, isolate and purify, test phosphorylation condition.In the presence of EGF, EGFR, Beclin1_WT and
Three kinds of mutation (Beclin1mutant A：229 tyrosine sites are only mutated by Beclin1_229E；Beclin1mutant
B：233 tyrosine sites are only mutated by Beclin1_233E；Beclin1mutant C：Beclin1_229E223E, pin
229 and 233 two tyrosine sites are mutated), phosphorylation test is carried out at 4 DEG C, 16 DEG C, 30 DEG C respectively, using exempting from
Epidemic disease blotting (Western Blot) is characterized, wherein, primary antibody is anti phosphotyrosine antibody.
Experimental result is as shown in Figure 2：Beclin1_WT tyrosine can by EGFR phosphorylations, and be influenced by temperature compared with
It is small；And then clear signal weakens by Beclin1_229E and Beclin1_233E, this shows that 229,233 the two sites are EGFR's
Substrate, and its sensitiveness temperature influence is obvious；And Beclin1_229E233E does not detect signal, show 229,233
The two sites or the main substrate site for EGFR.This result of the test illustrates that two sites are served as important is repaiied by structure
Decorations cause the role that function changes.
Clinical rectal neoplasm LC3-I/II, the Beclin1 aggregate level of embodiment 3 and phosphorylation level analysis
Five parts of rectal neoplasm clinical samples (III phases, operation obtains sample, numbering A, B, C, D, E) are obtained, Western blotting is used
Method (Western Blot) is characterized to LC3-I/II and Beclin1 aggregate levels in five parts of samples：I.e. first from paraffin organization
Corresponding destination protein is extracted, and is directly detected with corresponding antibody；Meanwhile, also to the Beclin1 phosphorylations in five parts of samples
Level is characterized：First carried out with anti-Beclin1 antibody after co-immunoprecipitation, then use anti phosphotyrosine antibody test.
Testing result is as shown in figure 3：As a result show, rectal neoplasm late period autophagy level and Beclin1 aggregate level it
Between without obvious relation between persistence.With reference to the clinical chemotherapy truth of five parts of samples, the phosphorylation that chemotherapeutics have impact on Beclin1 leads to
Cetuximab and the antibody that Victibix is targeting EGFR in the targeting combination of road, such as chemotherapy, it is to EGFR EGFR-TKs
The inhibitory action of activity make it that Beclin1 phosphorylation levels are lowered, so that autophagy suppression is released, so result is shown：Beclin1
Phosphorylation (p-Beclin1 in figure) level is lower, the increase of LC3-II band intensities.But autophagy enhancing is for anti-rectal neoplasm
Treatment is that profit is that disadvantage remains to be discussed, it has been found that the clinical evaluation of the sample D in Fig. 3 is not as good as sample C, this prompting Beclin1 phosphorus
Acidifying level can as Medication monitor reference.Therefore, Beclin1 expressions and its combine with p-Beclin1 ratio
Assess, the reference that can be predicted as Medication monitor, drug resistance.
Beclin1 expressions are analyzed in the clinical rectal neoplasm different tissues of embodiment 4
The tissue samples (III phases, operation obtains sample) of 20 clinical Patients With Rectal Carcinomas are chosen, and it is anti-with anti-Beclin1
Body carries out immunohistochemical experiment analysis to the rectal cancer tissue of each Patients With Rectal Carcinoma and cancer beside organism.
Its result is as shown in Figure 4：Knowable to figure, expression (Fig. 4 Bs of the Beclin1 in rectal cancer cell：Cancer group
Active immunity group result) apparently higher than expression (Fig. 4 A of carcinoma of the rectum cancer beside organism：Cancer beside organism's ImmunohistochemistryResults Results), this
Illustrate during developing from normal cell to rectal neoplasm, Beclin1 expression rise.Therefore, Beclin1 expression
The reference that level can be early diagnosed as rectal neoplasm.
The clinical anti-rectal neoplasm medicine autophagy Capability Categories of embodiment 5
The table of LC3-I/II and p62 in the Hela cells after anti-rectal neoplasm drug-treated is analyzed with immunoblot analysis
Up to level, the autophagy ability for rectal neoplasm medicine of being classified according to LC3-II conversion ratio and p62 removal rate.Detailed process is such as
Under：
First divide four groups by Hela cells to be cultivated, every group of transfection LC3 and p62, which enter after Hela cells, carries out excessive table
Reach；Wherein three groups respectively with conventional medicine (the Drug A of three class rectal neoplasm chemotherapy：Capecitabine；Drug B：Irinotecan；
Drug C：" oxaliplatin+fluorouracil+Calciumlevofolinate " is combined) Hela cells are handled, another set does blank control
(Control)；Four groups of Hela cells are incubated 24 hours in 37 DEG C of incubators, and by Hela cell PBSs, lysis
Buffer is cracked, and finally extracts cell protein.By the cell protein of extraction after denatured by boiling, entered using immunoblot experiment
Row target protein is characterized：Characterize LC3-II conversion ratio and p62 removal rate.
Final result is as shown in Figure 5：Test result indicate that, " oxaliplatin+fluorouracil+Calciumlevofolinate " combination group
(Drug C) relatively unused anti-rectal neoplasm drug-treated group (Control), its LC3-II conversion ratio is significantly improved, simultaneously
P62 albumen removal rate (degradation rate) is improved, that is, shows that Drug C have autophagy facilitation；And at capecitabine group (Drug A)
In Irinotecan group (Drug B), LC3-II conversion ratio and p62 removal rate have no significant change, i.e. Drug A and
Drug B do not have scanning quantitation after autophagy facilitation, development to analyze, and Drug C LC3-II conversion ratios are Drug A and Drug
Twice of B.With the classification using autophagy ability as standard is carried out to rectal neoplasm medicine excessively, clinical rectal neoplasm patient will be treated
There is provided and instruct.
Presently preferred embodiments of the present invention is the foregoing is only, is not intended to limit the invention, all essences in the present invention
Any modification, equivalent and improvement made within refreshing and principle etc., should be included within the scope of the present invention.
SEQUENCE LISTING
<110>Shenzhen people's benevolence biological medicine Science and Technology Ltd.
<120>Rectal neoplasm mark and its application, kit
<160> 1
<170> PatentIn version 3.5
<210> 1
<211> 171
<212> PRT
<213>Artificial sequence
<400> 1
Gly Pro Gly Ser Glu Ala Ser Asp Gly Gly Thr Met Glu Asn Leu Ser
1 5 10 15
Arg Arg Leu Lys Val Thr Gly Asp Leu Phe Asp Ile Met Ser Gly Gln
20 25 30
Thr Asp Val Asp His Pro Leu Cys Glu Glu Cys Thr Asp Thr Leu Leu
35 40 45
Asp Gln Leu Asp Thr Gln Leu Asn Val Thr Glu Asn Glu Cys Gln Asn
50 55 60
Tyr Lys Arg Cys Leu Glu Ile Leu Glu Gln Met Asn Glu Asp Asp Ser
65 70 75 80
Glu Gln Leu Gln Met Glu Leu Lys Glu Leu Ala Leu Glu Glu Glu Arg
85 90 95
Leu Ile Gln Glu Leu Glu Asp Val Glu Lys Asn Arg Lys Ile Val Ala
100 105 110
Glu Asn Leu Glu Lys Val Gln Ala Glu Ala Glu Arg Leu Asp Gln Glu
115 120 125
Glu Ala Gln Glu Gln Arg Glu Glu Ser Glu Phe Lys Arg Gln Gln Leu
130 135 140
Glu Leu Asp Asp Glu Leu Lys Ser Val Glu Asn Gln Met Arg Tyr Ala
145 150 155 160
Gln Thr Gln Leu Asp Lys Leu Lys Lys Thr Asn
165 170
Claims (10)
1. a kind of rectal neoplasm mark, it is characterised in that the rectal neoplasm mark is Beclin1 albumen.
2. for detecting the expression of Beclin1 albumen as claimed in claim 1 and/or the reagent of phosphorylation level in system
It is ready for use on detection and/or Diagnosis of Rectal tumour or in the treatment to rectal neoplasm patient and/or the kit of prognostic evaluation
Application.
3. application as claimed in claim 2, it is characterised in that the phosphorylation level of the Beclin1 albumen is described
229 sites of Beclin1 albumen and the tyrosine phosphorylation level in 233 sites.
4. application as claimed in claim 2, it is characterised in that the reagent bag of the expression of the detection Beclin1 albumen
The Beclin1 protein antibodies of anti-Beclin1 protein antibodies or fluorescence probe modification are included, the phosphoric acid of the Beclin1 albumen is detected
The reagent of change level includes anti phosphotyrosine antibody.
5. application as claimed in claim 2, it is characterised in that the expression and/or phosphorus of the described Beclin1 albumen of detection
The method of acidifying level is immunological method.
6. application as claimed in claim 4, it is characterised in that the immunological method includes Western blot, SABC
At least one in method and co-immunoprecipitation method.
7. application as claimed in claim 2, it is characterised in that the institute for the treatment of and/or prognostic evaluation to rectal neoplasm patient
State kit and anti-rectal neoplasm combination therapies；
Wherein, the anti-rectal neoplasm medicine includes capecitabine, Irinotecan, oxaliplatin, fluorouracil and Calciumlevofolinate
In at least one.
8. a kind of detection and/or Diagnosis of Rectal tumour or the treatment to intestinal tumor patient and/or the kit of prognostic evaluation, it is special
Levy and be, the kit includes the expression and/or phosphorylation water for detecting Beclin1 albumen as claimed in claim 1
Flat reagent.
9. as claimed in claim 8 in kit, it is characterised in that the phosphorylation level of the Beclin1 albumen is described
229 sites of Beclin1 albumen and the tyrosine phosphorylation level in 233 sites.
10. kit as claimed in claim 8, it is characterised in that the reagent of the expression of the detection Beclin1 albumen
The Beclin1 protein antibodies modified including anti-Beclin1 protein antibodies or fluorescence probe, detect the phosphorus of the Beclin1 albumen
The reagent of acidifying level includes anti phosphotyrosine antibody.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710247473.5A CN106950384A (en) | 2017-04-14 | 2017-04-14 | Rectal neoplasm mark and its application, kit |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201710247473.5A CN106950384A (en) | 2017-04-14 | 2017-04-14 | Rectal neoplasm mark and its application, kit |
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| Publication Number | Publication Date |
|---|---|
| CN106950384A true CN106950384A (en) | 2017-07-14 |
Family
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201710247473.5A Pending CN106950384A (en) | 2017-04-14 | 2017-04-14 | Rectal neoplasm mark and its application, kit |
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| Country | Link |
|---|---|
| CN (1) | CN106950384A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109557317A (en) * | 2019-01-10 | 2019-04-02 | 南方医科大学南方医院 | Application of the ATXN2L as the marker of aided assessment gastric cancer oxaliplatin secondary resistance |
| WO2020024239A1 (en) * | 2018-08-03 | 2020-02-06 | 暨南大学 | Method for evaluating efficacy of drug for reversal of multi-drug resistance of tumors |
| WO2021185234A1 (en) * | 2020-03-16 | 2021-09-23 | 正大天晴药业集团股份有限公司 | Combined pharmaceutical composition of compound as c-met kinase inhibitor and use thereof |
| CN113804885A (en) * | 2020-06-11 | 2021-12-17 | 复旦大学 | Marker for detecting early tumors and application thereof |
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| CN101971033A (en) * | 2007-06-04 | 2011-02-09 | 戴诺普雷克斯公司 | Biomarker combinations for colorectal cancer |
| WO2014046966A1 (en) * | 2012-09-24 | 2014-03-27 | Board Of Regents, The University Of Texas System | Beclin 1 phosphorylation |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101971033A (en) * | 2007-06-04 | 2011-02-09 | 戴诺普雷克斯公司 | Biomarker combinations for colorectal cancer |
| WO2014046966A1 (en) * | 2012-09-24 | 2014-03-27 | Board Of Regents, The University Of Texas System | Beclin 1 phosphorylation |
Non-Patent Citations (2)
| Title |
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| BECLIN1、LIN28B在结直肠腺瘤中的表达: "张东姣", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
| 张东姣: ""Beclin1、lin28b在结直肠腺瘤中的表达", 《中国优秀硕士学位论文全文数据库 医药卫生科技辑》 * |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2020024239A1 (en) * | 2018-08-03 | 2020-02-06 | 暨南大学 | Method for evaluating efficacy of drug for reversal of multi-drug resistance of tumors |
| CN109557317A (en) * | 2019-01-10 | 2019-04-02 | 南方医科大学南方医院 | Application of the ATXN2L as the marker of aided assessment gastric cancer oxaliplatin secondary resistance |
| WO2021185234A1 (en) * | 2020-03-16 | 2021-09-23 | 正大天晴药业集团股份有限公司 | Combined pharmaceutical composition of compound as c-met kinase inhibitor and use thereof |
| CN115135326A (en) * | 2020-03-16 | 2022-09-30 | 正大天晴药业集团股份有限公司 | Combined pharmaceutical composition of compounds as c-Met kinase inhibitor and application thereof |
| CN113804885A (en) * | 2020-06-11 | 2021-12-17 | 复旦大学 | Marker for detecting early tumors and application thereof |
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Application publication date: 20170714 |