CN106383235A - Application of breast cancer multidrug resistant protein molecule IBP - Google Patents
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Abstract
The invention discloses an application of a breast cancer multidrug resistant protein molecule IBP, wherein the IBP has an amino acid sequence as shown in SEQ ID NO.11; studies find out that expression of the IBP in breast cancer multidrug resistant cells MCF-7/ADR is significantly higher than expression in MCF-7 parental cells, and moreover, after the expression of the IBP in the breast cancer cells is decreased, the sensibility on a variety of chemotherapy drugs is significantly enhanced; after the expression of the IBP in the MCF-7/ADR is interfered, the breast cancer drug-resistant cells can be inhibited, besides, expression and drug transport functions of MDR1 and other multidrug resistant related genes in the MCF-7/ADR are inhibited, and the drug sensitivity is increased; therefore, the IBP can be used as a multidrug resistant target gene for detection of breast cancer, and also can be used for preparing breast cancer multidrug resistant preparations or drugs.
Description
Technical field
The invention belongs to biochemical field is and in particular to the application of breast carcinoma Mdr-p matter molecule IBP.
Background technology
Breast carcinoma is the major disease seriously threatening WomanHealth, be lead to women die second largest cancer related because
Element.Studies have found that, although facilitating chemotherapy, still have quite a few patient that transfer and the recurrence of breast carcinoma occur.
According to statistics, 5 years survival rates of metastatic breast cancer only have 26.7%.The metastatic breast cancer patient having 90% is due to chemotherapeutic
Thing tolerance leads to Endodontic failure.For metastatic breast cancer patient, the progressive antitumor drug drug resistance in treatment is one
Individual major issue.Although importantly, clinic has multiple antitumor drug, once occurring a kind of medicine resistance in the course for the treatment of
It is subject to, most of medicines no longer will produce response to treatment to patient, and this phenomenon is referred to as multidrug resistance.
Existing achievement in research shows, take part in the multidrug resistance of breast carcinoma from the many factors of breast carcinoma itself, main
Including：Medicine flows out mechanism correlation multidrug resistance gene such as MDR1, MRP1, ABCG2 etc. expression and changes, and other non-cross-film machines
The atypia drug resistance approach of system mediation.But the breast carcinoma resistance currently for these mechanism reverses achievement in research in clinical practice
Effect in application is unsatisfactory.
Content of the invention
In view of this, an object of the present invention is to provide breast carcinoma Mdr-p matter molecule IBP as diagnosis
Application in breast carcinoma multidrug resistance mark；The second object of the present invention is to provide detection breast carcinoma Mdr-p matter
Application in preparing Diagnosis of Breast cancer multidrug resistance test kit for the reagent of molecule IBP；The third object of the present invention is to provide
The reagent reducing breast carcinoma Mdr-p matter molecule IBP expression is applied in preparing anti-breast cancer multidrug resistance preparation.
For achieving the above object, the present invention provides following technical scheme：
1st, breast carcinoma Mdr-p matter molecule IBP is as the application in Diagnosis of Breast cancer multidrug resistance mark, institute
The aminoacid sequence stating breast carcinoma Mdr-p matter molecule IBP is as shown in SEQ ID NO.11.
2nd, Diagnosis of Breast cancer multidrug resistance test kit prepared by the reagent of detection breast carcinoma Mdr-p matter molecule IBP
In application, the aminoacid sequence of described breast carcinoma Mdr-p matter molecule IBP is as shown in SEQ ID NO.11.
3rd, anti-breast cancer multidrug resistance preparation prepared by the reagent reducing breast carcinoma Mdr-p matter molecule IBP expression
Middle application, the aminoacid sequence of described breast carcinoma Mdr-p matter molecule IBP is as shown in SEQ ID NO.11.
Further, the reagent of described reduction breast carcinoma Mdr-p matter molecule IBP expression is interference breast carcinoma multiple medicines
The carrier of drug-resistant protein matter molecule IBP expression or medicine.
Further, the reagent of described reduction breast carcinoma Mdr-p matter molecule IBP expression is interference breast carcinoma multiple medicines
The carrier of drug-resistant protein matter molecule IBP expression.
Further, the disturbance target point such as SEQ of the carrier of described interference breast carcinoma Mdr-p matter molecule IBP expression
Shown in ID NO.12 or SEQ ID NO.13.
The beneficial effects of the present invention is：The present invention have studied breast carcinoma Mdr-p matter molecule IBP in breast carcinoma
Overexpression in multidrug resistance cell, in breast cancer medicines sensitive cellss low expression characteristic, IBP and breast cancer cell multiple medicines
Drug resistance is closely related, and the breast cancer cell after overexpression IBP has been directed to the drug resistance ability of multiple breast cancer treatment common drugs
Strengthen, the obvious up-regulated of multidrug resistance gene MDR1；Expression in breast carcinoma MCF-7/ADR mdr cell for the IBP is bright
Aobvious increase, interference expression in breast carcinoma MCF-7/ADR mdr cell for the IBP can significantly improve its sensitivity to doxorubicin
Property hence it is evident that reducing the expression of multidrug resistance gene MDR1 etc., lower the drug efflux ability of breast carcinoma resistance cell, for breast
Adenocarcinoma multidrug resistance provides a kind of new mark.And on this basis, in order to design and to develop Diagnosis of Breast cancer multiple medicines resistance to
Medicine test kit, it is directed to the new drug of breast carcinoma multidrug resistance.
Brief description
In order that the purpose of the present invention, technical scheme and beneficial effect are clearer, the present invention provides drawings described below：
Fig. 1 is breast carcinoma multiple medicine-resistant cell line MCF-7/ADR drug resistance ability and multidrug resistance gene MDR1
(A represents that CCK-8 detects MCF-7/ADR and its parental cell MCF-7 48 hours under the doxorubicin effect of variable concentrations for expression
Cell survival rate afterwards；B represents that MCF-7/ADR and MCF-7 acts on the half-inhibition concentration IC50 ratio of 48 hours in doxorubicin
Relatively；C represents the expression of fluorescence quantitative PCR detection MCF-7/ADR and MDR1 and IBP in MCF-7；D represents that Western detects
The expression of MDR1 and IBP in MCF-7/ADR and MCF-7).
Fig. 2 be after IBP interference expression in MCF/ADR cell the expression of IBP and multidrug resistance gene (A represents
Fluorescence quantitative PCR detection IBP and the expression of multidrug resistance gene；B represents Western detection IBP and multidrug resistance
The expression of associated protein).
For cellular drug resistance after MCF-7/ADR interference IBP expression, (A represents interference in CCK-8 detection MCF-7/ADR to Fig. 3
After IBP expression under the doxorubicin effect of variable concentrations the cell survival rate after 48 hours；B represents interference in MCF-7/ADR
The half-inhibition concentration IC50 acting on 48 hours in doxorubicin after IBP expression compares).
Fig. 4 is for Western detection MCF-7/ADR interference IBP expression group and its compared with control cells in variable concentrations doxorubicin
Apoptosis situation under effect.
For colony formation, Fig. 5 detects that (A represents that MCF-7/ADR interference IBP expression group and its compared with control cells are dense in difference
Cell proliferative conditions under degree doxorubicin effect；B represents IBP overexpression MCF-7 cell and its compared with control cells in variable concentrations
Cell proliferative conditions under doxorubicin effect).
Fig. 6 is that (A represents before outer row Rh123 using outer row's ability after the interference IBP expression of flow cytometer showed MCF-7/ADR cell
Afterwards in cell Rh123 ratio, left for before outer row, right for after outer row；B represents the discharge rate of Rh123).
Fig. 7 is the expression (A of IBP and MDR1 after overexpression IBP in MCF-7 cell：Fluorescence quantitative PCR detection
The expression of IBP and MDR1 after overexpression IBP in MCF-7 cell；B：Overexpression IBP in Western detection MCF-7 cell
The expression of IBP and MDR1 afterwards.
Fig. 8 is phonetic in doxorubicin (DOX), paclitaxel (PTX), 5 '-fluorine urine for CCK8 detection IBP overexpression MCF-7 cell
Drug resistance in pyridine (5 '-FU), cisplatin (DDP).
Fig. 9 detects the shRNA skill using lentivirus mediated in breast carcinoma cell strain SK-BR-3 and MDA-MB-231 for CCK8
Art reduces the drug resistance after IBP expresses in doxorubicin (DOX), paclitaxel (PTX).
Specific embodiment
Below in conjunction with accompanying drawing, the preferred embodiments of the present invention are described in detail.Unreceipted concrete in embodiment
The experimental technique of condition, generally according to normal condition, such as Molecular Cloning:A Laboratory guide (third edition, J. Pehanorm Brooker etc. writes)
Described in condition, or according to the condition proposed by manufacturer.
The culture of embodiment 1, the induction of breast carcinoma resistance cell strain MCF-7/ADR and breast carcinoma cell strain
MCF-7, MDR-MB-231, SK-BR-3 breast carcinoma cell strain is purchased from U.S.'s ATCC cell bank, is voluntarily frozen by this room
Deposit.MCF-7 cell culture in 1640 culture medium containing 10% (v/v) hyclone, the MCF-7/ADR cell of multidrug resistance by
MCF-7 parental cell is constantly exposed to induce in the doxorubicin culture medium of gradient concentration and forms, wherein doxorubicin concentration by
5ng/ml is gradually increased to 1000ng/ml.Using doxorubicin containing 1000ng/ml, 10% tire cattle during MCF-7/ADR long-term cultivation
1640 culture medium of serum, in 37 DEG C, the CO containing 5% (v/v)2Culture.MDR-MB-231, SK-BR-3 cell culture is in containing
The DMEM culture medium of 10% (v/v) hyclone.
The doxorubicin of the MCF-7/ADR cell variable concentrations of the multidrug resistance building is processed, adopts after 48 hours
CCK-8 detects cell survival rate, and result is as shown in A in Fig. 1.Result shows, MCF-7/ADR makees in the doxorubicin of variable concentrations
Higher with lower survival rate.Then compare the half suppression that MCF-7/ADR and MCF-7 acts on 48 hours in 1000ng/ml doxorubicin
Concentration IC50 processed, result is as shown in B in Fig. 1.Result shows, the half-inhibition concentration IC50 of MCF-7/ADR is higher than MCF-7, its
Resistance index is 57.48.
Then Realtime-PCR is adopted to detect MDR1 gene and IBP gene in MCF-7/ADR cell and MCF-7 cell
Expression, comprises the following steps that：Each cell culture collected cell after 24 hours, according to RNA extracts kit Trizol step,
Extract total serum IgE, and measure concentration and the quality of RNA with ultraviolet spectrophotometer.Then adopt PrimeScriptTMRT
Reagent Kit random primer reverse transcription RNA obtains cDNA, and reverse transcription system is as follows：5 × PrimeScript buffer 4 μ l,
PrimeScript Enzyme Mix 1 μ l, Oligo dT Primer (50 μM) 1 μ l, Random 6mers (100 μM) 4 μ l,
RNA 1μg；Reaction condition：37 DEG C of incubations 15min, 85 DEG C of enzyme-deactivating 5s；Adopt 2 × Real-time PCR Master again
MIX, with reversion rate product as masterplate, SYBR green method expands.Reaction system is as follows：ddH2O 2 μ l, 2 × Real-time
PCR Master MIX 5 μ l, forward primer 0.5 μ l, downstream primer 0.5 μ l, cDNA2 μ l.Reaction condition：95 DEG C of reaction 1min,
47 are circulated in 95 DEG C, 5s；58 DEG C, 5s；72 DEG C, 20s；Gather fluorescence, solubility curve when 72 DEG C, rise to 95 DEG C of mistakes from 60 DEG C
Cheng Zhong, each circulation increases by 0.5 DEG C, totally 71 circulations, and each PCR reaction primer sequence is shown in Table 1.
Table 1, fluorescence quantification PCR primer sequence
Quantitative result is calculated using Comparative Delta-delta Ct method, experiment is minimum in triplicate, to each thin
Each in born of the same parents' strain
Mrna expression amount carries out statistical analysiss, and result is as shown in C in Fig. 1.Result shows, IBP is thin in MCF-7/ADR multiple medicines
Expression in born of the same parents' drug-resistant cell strain substantially increases compared with MCF-7 parental cell strain.
Then Western Blot is adopted to detect the expression feelings in MCF-7/ADR cell and MCF-7 cell for MDR1 and IBP
Condition, concrete grammar is as follows：Scrape cell with cell after adding RIPA cell pyrolysis liquid, after fully cracking on ice in rotating speed be
It is centrifuged 5min under the conditions of 14000g, take supernatant, abandon precipitation.BCA method measures sample protein mass concentration in supernatant, is subsequently adding suitable
Amount 5 × sample-loading buffer, boils -80 DEG C of preservations after 5min；Again by cell protein with 30 μ g/ hole rows PAGE gel electrophoresis,
Constant current 0.25A, transferring film 180min, after pvdf membrane closing, overnight (one anti-is respectively anti-IBP (this laboratory for 4 DEG C of an anti-incubation
Provide for oneself, 1：1000), anti-MDR1 (Santa Cruz, 1：1000), anti-PARP (Cell Signaling
Technology, 1：1000), anti-ABCG2 (Cell Signaling Technology, 1：1000), anti-MRP
(Santa Cruz, 1：1000), anti-Tublin (Cell Signaling Technology, 1：5000), anti-GAPDH
(Cell Signaling Technology, 1：5000)), TBST washs 4 times, each 15min, then uses HRP labelling goat to resist
Mus/rabbit igg (1：10000) (18～25 DEG C) incubation 1h of room temperature, TBST wash 6 times, and each 15min adds ECL reagent, film
Exposure tests, result is as shown in D in Fig. 1.Result shows, expression in MCF-7/ADR multiple medicines cells resistance cell strain for the IBP is relatively
MCF-7 parental cell strain substantially increases.
MCF-7/ADR, MDA-MB-231, SK-BR-3 that embodiment 2, the IBPshRNA of structure lentivirus mediated disturb are steady
Determine cell line
By bioinformatic analysis, for IRF4 associated proteins (IBP) gene (Genbank：NM_022047), its ammonia
Base acid sequence as shown in SEQ ID NO.11, then 2 shot design RNA interference sequences according to IBP gene, target sequence and
Interference sequence is as shown in table 2.Synthetic oligo is annealed into double-strand oligo sequence, connects and disturb load into linearizing RNA
Body LV10, obtains the LV10 carrier containing aim sequence.The correct transformant of sequence verification, carries out high-purity plasmid extraction.Then
The LV10 carrier containing aim sequence building is entered with shuttle plasmid and packaging plasmid (pGag/Pol, pRev, pVSV-G) cotransfection
293T cell, collects cells and supernatant, ultracentrifugation concentrating virus, -80 DEG C of preservations.
Table 2.IBP gene target and RNA interference sequence
Well-grown MCF-7/ADR, MDA-MB-231 and SKBR-3 cell is taken to be seeded to 24 orifice plates, every hole total cellular score
About 5 × 104Individual, in 37 DEG C, containing 5% (v/v) CO2Culture 24 hours.Then virus is taken to infect MCF-7/ADR, MDA-MB- respectively
231 and SKBR-3 cells, infection MCF-7/ADR cell is 5 using MOI value, without cohesion amine (polybrene)；Infection
MDA-MB-231 and SKBR-3 cell is 10 using MOI value, with complete medium virus dilution liquid and add final concentration of 5 μ g/
The Polybrene of ml, persistent virus infections change fresh complete medium after 18 hours.Treat that cell covers with culture hole, do 1：
Move into culture bottle culture after 10 dilutions, and add puromycin (Puromycin) to be screened.Screening time is no less than 14 days,
Period cell normally passes on and changes liquid, obtains stable infection cell strain.
Method according to embodiment 1 is passed through quantitative fluorescent PCR and is detected that IBP, MDR1, MRP are related with ABCG2 with Western
The expression of multidrug resistance gene, result is as shown in Figure 2.Result shows, after lentivirus mediated IBPA interference expression, related multiple medicines is resistance to
Medicine gene all reduces in gene level and protein expression amount.
Then the MCF-7/ADR cell of infection is adopted CCK8 to detect survival rate, the MCF-7/ of the IBP to be uninfected by simultaneously
, as comparison, result is as shown in A in Fig. 3 for ADR cell.Result shows, in variable concentrations after interference IBP expression in MCF-7/ADR
Doxorubicin act on lower 48 hours after cell survival rate be less than matched group.Then detect the MCF-7/ADR cell half of infection
Number inhibition concentrations IC50, simultaneously using the MCF-7/ADR cell of IBP that is uninfected by as comparison, result is as shown in B in Fig. 3.Result
Display, the half-inhibition concentration IC50 acting on 48 hours in doxorubicin after interference IBP expression in MCF-7/ADR cell relatively compares
Group is low.
Utilize after Western detection MCF-7/ADR cell interference IBP expression in variable concentrations according to above-mentioned identical method
Apoptosis situation under doxorubicin effect, result is as shown in Figure 4.Result shows, the interference IBP expression of MCF-7/ADR cell
Afterwards under the conditions of 5 μ g/ml, 20 μ g/ml doxorubicins stimulate 24 hours, IBP low expression group apoptosis mark Cleaved-
PARP expression substantially increases, and points out the expression of interference IBP can promote drug-induced apoptosis.
Act in variable concentrations doxorubicin using after colony formation detection MCF-7/ADR cell interference IBP expression
Under cell proliferative conditions, comprise the following steps that：Take the logarithm each group cell of trophophase, use mass fraction 0.25% pancreas egg respectively
White enzymic digestion simultaneously blows and beats into individual cells, and cell suspension in the corresponding complete culture solution of hyclone of volume fraction 10%
Standby；Cell suspension is made gradient multiple dilutions, MCF-7/ADR NC, siIBP2# group cell is with 300 cells in every hole, MCF-
7/C1, MCF-7/IBP group cell inoculates six orifice plates with 500 cells in every hole, and gently rotates, and so that cell is uniformly dispersed.Put 37
℃、CO2Volume fraction be 5% cell culture incubator in overnight incubation.Each hole adds the how soft ratio of corresponding variable concentrations within second day
Star, continues 37 DEG C, 5%CO2Culture changed liquid after 24 hours, persistently cultivated 2～3 weeks.Often observe, when appearance naked eyes in culture dish
During visible clone, terminate culture.Abandoning supernatant, carefully embathes 2 times with PBS；Plus the paraformaldehyde of mass fraction 4% fixes
Cell fixes 15 minutes.Then remove fixative, plus appropriate GIMSA application dyeing liquor contaminates 30 minutes, is then slowly washed away with flowing water
Dyeing liquor, air is dried.Plate is inverted and is superimposed a transparent film with grid, with the naked eye directly count clone, or
Microscope (low power lens) counts the clone's number more than 10 cells.Finally calculate cloning efficiency, cloning efficiency=(clone
Number/inoculating cell number) × 100%, result is as shown in A in Fig. 5.Result shows, after the interference IBP expression of MCF-7/ADR cell,
1 μ g/ml, 2 μ g/ml, 3 μ g/ml doxorubicins act on 24 hours after ability of cell proliferation all substantially weaken.
Drug efflux ability after flow cytometry detection MCF-7/ADR cell interference IBP expression, comprises the following steps that：
Become 1 × 10 with the pancreatin digestion corresponding cell preparation of mass fraction 0.25%6/ ml concentration single cell suspension, every kind of cell with
1ml adds 6 orifice plates, overnight incubation, treats cell attachment.Every hole adds the Rh123 of 5.0 μ g/ml, continues culture 60min, digests,
1000g is centrifuged 5min, supernatant discarded, and ice-cold 0.01M PBS washes cell 2 times, take 250 μ l to arrange outside flow cytomery before
Rh123 average fluorescent strength.In test tube, remaining cell continues to add cellar culture 60min in culture medium culturing plate, and 1000g is centrifuged
5min, supernatant discarded, ice-cold 0.01M PBS washes cell 2 times, takes 250 μ l average with Rh123 after arranging outside flow cytomery
Fluorescence intensity × 100%；Flow cytometer measures Rh123 fluorescence intensity at excitation wavelength 488nm, launch wavelength 525nm, meter
Number about 2 × 104Individual cell；Before drug efflux efficiency=(average fluorescent strength after average fluorescent strength-outer row before arranging outward)/outer row
Average fluorescent strength, result is as shown in Figure 6.Result shows, after IBP expression reduction, MCF/ADR cell is to medicine efflux pump substrate
The discharge rate of Rh123 substantially reduces.
Embodiment 3, the MCF-7 stably transfected cell line of structure IBP overexpression
Extract human lymphocyte total serum IgE, its reverse transcription is cDNA.With human lymphocyte cDNA as template, using PCR skill
Art expands people's IBP full-length cDNA fragment.People's IBP full-length cDNA fragment is cloned in carrier for expression of eukaryon pEGFP-C1, through surveying
Sequence is identified, successfully builds pEGFP fusion expression plasmid pEGFP-C1-IBP.By 1 μ g pEGFP-C1 and pEGFP-C1-IBP matter
Grain, transfects the MCF-7 cell in 24 orifice plates respectively, obtains IBP overexpression MCF-7 cell.Transfection adopts
Lipofectamine2000 test kit；By cell dissociation after transfecting 24 hours, then with volume ratio for 1：24 pairs of cell suspension enter
Row dilution is inoculated in 24 new orifice plates, continues culture, cell culture condition MCF-7 cell culture condition as previously mentioned；Training
Add 1000 μ g/ml G418 (Geneticin, Geneticin) in the medium after supporting 24 hours；After screening and culturing 10 days, disappear
Change cell in the culture hole that fluorecyte ratio is high, fluorescence intensity is bright, after counting, cell is diluted in 96 orifice plate Colony Culture；
The good single clone's amplification culture of fluorescence intensity high state is selected in 96 orifice plates of cloning.
Then according to the method for embodiment 1 utilize quantitative fluorescent PCR and Western detection transfection MCF-7 cell IBP and
The expression of MDR1, result is as shown in Figure 7.Result shows, and transfects IBP overexpression plasmid in breast cancer cell MCF-7
Afterwards, IBP and MDR1 expression increases, and that is, drug resistance in multi-medicament for the MCF-7 is remarkably reinforced.
IBP overexpression MCF-7 cell is processed under variable concentrations doxorubicin, simultaneously to transfect the MCF-7 of empty plasmid
Cell is comparison, then utilizes colony formation to detect cell proliferative conditions, result is as shown in B in Fig. 5.Result shows, IBP
Overexpression MCF-7 cell cell after concentration is 0.1 μ g/ml and 0.2 μ g/ml doxorubicin acts on 24 hours compared with compared with control cells
Multiplication capacity all strengthens.
Embodiment 4, cell proliferation-toxicity test detection different pharmaceutical processes lower cell inhibitory rate
Well-established IBP overexpression MCF-7 cell strain amplification culture is good to cell state, each thin with pancreatin digestion
Born of the same parents, and with, after neutralization pancreatin digestion, washing cell once rear cell counting with aseptic PBS, each thin using complete medium adjustment
Born of the same parents' concentration is 5 × 104Then the cell suspension adjusting concentration be placed in the Tissue Culture Dish of aseptic 10cm, in front and back by/ml
Left and right jog mixes cell, adds in 96 orifice plates using the volley of rifle fire by uniform for cell suspension, every hole 100 μ l, 37 DEG C of overnight incubation,
Add the medicine of corresponding variable concentrations after cell attachment, the cycle hole of 96 orifice plates is not added with cell suspension, plus aseptic PBS in case
Culture medium is evaporated；Then change liquid with the volley of rifle fire, each hole adds and accordingly contains variable concentrations doxorubicin respectively with gradient dilution
(DOX), paclitaxel (PTX), 5'-FU (5 '-FU), the complete medium of cisplatin (DDP), and also add in acellular hole
Enter the pastille culture medium of corresponding variable concentrations, as acellular blank, every hole adds 100 μ l pastille complete mediums, often
Individual drug level at least does 5 multiple holes, 37 DEG C of cultures, and after drug treating terminates, every hole adds 10 μ L CCK-8 solution, in cell
Continue culture 1 hour, 450nm mensuration absorbance, experimental result is represented with cell survival rate in incubator：Cell survival rate (%)
=(drug treating group-relative medicine concentration blank)/(non-drug treating group-blank) × 100%, result such as Fig. 8
Shown.Result shows, after overexpression IBP, the survival rate of MCF-7 cell dramatically increases, and shows MCF-7 cell after IBP overexpression
Drug resistance is remarkably reinforced.
Then according to MDA-MB-231 the and SKBR-3 cell that the interference IBP that embodiment 1 is obtained is expressed by identical method
Cell survival rate under doxorubicin (DOX) and paclitaxel (PTX) effect, result is as shown in Figure 9.Result shows, in SK-BR-3
After interference IBP expression in MDA-MB-231 cell, two kinds of cells of SK-BR-3 and MDA-MB-231 are in doxorubicin and paclitaxel
In sensitivity substantially increase.
Finally illustrate, preferred embodiment above only in order to technical scheme to be described and unrestricted, although logical
Cross above preferred embodiment the present invention to be described in detail, it is to be understood by those skilled in the art that can be
In form and various changes are made to it, without departing from claims of the present invention limited range in details.
Claims (6)
1. breast carcinoma Mdr-p matter molecule IBP is as the application in Diagnosis of Breast cancer multidrug resistance mark, described breast
The aminoacid sequence of adenocarcinoma Mdr-p matter molecule IBP is as shown in SEQ ID NO.11.
2. the reagent of detection breast carcinoma Mdr-p matter molecule IBP is in preparing Diagnosis of Breast cancer multidrug resistance test kit
Application, the aminoacid sequence of described breast carcinoma Mdr-p matter molecule IBP is as shown in SEQ ID NO.11.
3. the reagent reducing breast carcinoma Mdr-p matter molecule IBP expression should in preparing anti-breast cancer multidrug resistance preparation
With the aminoacid sequence of described breast carcinoma Mdr-p matter molecule IBP is as shown in SEQ ID NO.11.
4. according to claim 3 application it is characterised in that：Described reduction breast carcinoma Mdr-p matter molecule IBP
The reagent of expression is carrier or the medicine of interference breast carcinoma Mdr-p matter molecule IBP expression.
5. according to claim 4 application it is characterised in that：Described reduction breast carcinoma Mdr-p matter molecule IBP
The reagent of expression is the carrier of interference breast carcinoma Mdr-p matter molecule IBP expression.
6. according to claim 4 application it is characterised in that：Described interference breast carcinoma Mdr-p matter molecule IBP
The disturbance target point of the carrier of expression is as shown in SEQ ID NO.12 or SEQ ID NO.13.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108950002A (en) * | 2018-09-06 | 2018-12-07 | 广东省中医院（广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院） | Application and kit based on Caveolin-1 detection chemosensitivity of breast cancer |
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2016
- 2016-08-31 CN CN201610789324.7A patent/CN106383235B/en not_active Expired - Fee Related
Non-Patent Citations (3)
| Title |
|---|
| A. A. STAVROVSKAYA: "Cellular Mechanisms of Multidrug Resistance of Tumor Cells", 《BIOCHEMISTRY》 * |
| MINGZHEN YANG ET AL: "Interferon regulatory factor 4 binding protein is a novel p53 target gene and suppresses cisplatin-induced apoptosis of breast cancer cells", 《MOLECULAR CANCER》 * |
| SC LINN ET AL: "p53 and P-glycoprotein are often co-expressed and are associated with poor prognosis in breast cancer", 《BRITSH JOURNAL OF CANCER》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108950002A (en) * | 2018-09-06 | 2018-12-07 | 广东省中医院（广州中医药大学第二附属医院、广州中医药大学第二临床医学院、广东省中医药科学院） | Application and kit based on Caveolin-1 detection chemosensitivity of breast cancer |
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| CN106383235B (en) | 2018-03-30 |
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