CN109490543A - It is a kind of predict gastrointestinal stromal tumor drug resistance kit and its application - Google Patents
It is a kind of predict gastrointestinal stromal tumor drug resistance kit and its application Download PDFInfo
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Abstract
The present invention relates to a kind of kits for predicting gastrointestinal stromal tumor drug resistance, kit includes Rad51 albumen, KU70 protein immunization group detection reagent and specification, and kit application method is: the normal tissue of patient and tumor tissues first being taken to carry out Rad51 albumen and KU70 protein immunization histochemical staining respectively；Pathological section read tablet is carried out afterwards, measures staining power and stained area；Immunohistochemistry scoring is carried out according to the calculation method of staining power x stained area, if result are as follows: (tumor tissues Rad51 score/normal tissue Rad51 score)>1, and when (tumor tissues KU70 score/normal tissue KU70 score)<1, then gastrointestinal stromal tumor is not in drug resistance, if result are as follows: (tumor tissues Rad51 score/normal tissue Rad51 score)<1, and when (tumor tissues KU70 score/normal tissue KU70 score)>1, then gastrointestinal stromal tumor will appear drug resistance.
Description
Technical field
The present invention relates to pharmaceutical technology fields, specifically, be it is a kind of predict gastrointestinal stromal tumor drug resistance kit and
It is applied.
Background technique
Gastrointestinal stromal tumor (GastrointestinalStromalTumors, GIST) is a kind of originating between gastrointestinal tract
The tumour of leaf texture accounts for the major part of leaf tumour between alimentary canal, and since tumour is insensitive to conventional radiotheraphy, operation excision was once controlled
The only effective method of GIST is treated, the patient of operative chance is lost for tumour progression transfer or postoperative recurrence, wherein position is raw
Deposit the phase only 6-18 months, survival rate is less than 10% within 5 years.10 Yu Nianlai, as modern molecular biology technique is examined in GIST clinic
Extensive use in disconnected treatment especially as the appearance of the molecular targeted agents of representative and is popularized using Imatinib, makes GIST patient
Prognosis significantly improve.Targeted therapy become oncotherapy hot topic, currently, Imatinib for treat advanced GIST and
The significant curative effect of GIST postoperative adjuvant therapy to be known together extensively, but the primary and secondary problems of Imatinib simultaneously
Also it is increasingly subject to researcher's concern, becomes the hot spot of recent GIST research.Currently, Imatinib and Sutent are for treating
GIST's is predominantly targeting drug, and Imatinib, tyrosine kinase inhibitor is a kind of small molecular protein kinase inhibitor, it has
Play the role of blocking one or more protein kinases.It is clinically used for treatment chronic myelogenous leukemia and malignant gastrointestinal interstitial is swollen
Tumor.Sutent can be used for treatment failure or intolerable gastrointestinal stromal tumor (GIST), inoperable advanced renal cell carcinoma
(RCC), unresectable, metastatic differentiated progressive stage pancreas neuroendocrine tumors (pNET) adult patients.Imatinib and
Sutent can act as the tyrosine kinase sites of counterfeit substrate Yu ATP competitive binding KIT/PDGFRA receptor, play retardance
The effect of kinase activator and signal transduction, in addition, Sutent also has the function of anticancer drug screening assay, although targeted drug
Treatment significantly improve the prognosis of GIST, about 80% patient benefits after initially receiving treatment with imatinib, but targets medicine
Object drug resistance is inevitable, and resistance problems become the thorny problem in targeted therapy.The study found that there is the trouble of 40%-50%
There is drug resistance in 2 years after receiving treatment with imatinib in person, and the patient that imatinib-resistant occurs such as receives Sutent treatment,
Its progression free survival phase.Drug resistance can be divided into primary drug resistance and secondary resistance, the former refer to after medication disease be still in progress or
Less than 6 months persons of stable disease, the latter refer to the progression of disease occurred again after treatment stable disease 6 months.Drug resistance
It may be related with following factor:
(1) gene mutation factor；
(2) overexpression etc. of KIT/PDGFRA gene.
Therefore, for the prediction of drug resistance of tumor, it can help to the selection of targeted drug.Have for the recovery of patient very big
Help.
Chinese patent application: CN105074009B is disclosed and a kind of prediction technique of the sensibility of EGFR inhibitor,
It include: that (a) determines process, which whether there is KRAS gene source to determine from the blood sample acquired in subject
Nucleic acid or its protein, and determine the blood sample in above-mentioned KRAS gene source nucleic acid or its protein be wild
Type or anomaly；And (b) judgment step, in above-mentioned operation (a), if detecting wild type in above-mentioned blood sample
The nucleic acid or its protein of the nucleic acid of KRAS gene source or its protein without detecting anomaly KRAS gene source,
A possibility that tumour of above-mentioned subject is sensitive to EGFR inhibitor height is then judged, if detecting variation in above-mentioned blood sample
The nucleic acid or its protein of type KRAS gene source, then judge the tumour of above-mentioned subject it is insensitive to EGFR inhibitor can
It can property height.But it yet there are no report about a kind of kit for predicting gastrointestinal stromal tumor drug resistance of the present invention and its application.
Summary of the invention
The first purpose of this invention is to provide a kind of prediction gastrointestinal stromal tumor drug resistance for limit the deficiencies in the prior art
Kit.
Second object of the present invention is in view of the deficiencies of the prior art, to provide a kind of prediction gastrointestinal stromal tumor drug resistance
The application of kit.
To realize above-mentioned first purpose, the technical solution adopted by the present invention is that:
A kind of kit for predicting gastrointestinal stromal tumor drug resistance, the kit includes operational manual, and the operation is said
Bright secretary is loaded with:
The score computation method are as follows: staining power * stained area；
It further include that normal tissue sample, Rad51 protein immunization group detection reagent and KU70 albumen are exempted from the kit
Epidemic disease group detection reagent, the Rad51 protein immunization group detection reagent include rabbit-anti people Rad51 polyclonal antibody, Rad51 egg
The white goat antirabbit fluorescence secondary antibody for extracting reagent, dyestuff, FITC or Dylight549 label；The KU70 protein immunization groupization inspection
Test agent includes the mountain of the anti-human KU70 monoclonal antibody of mouse, KU70 Protein Extraction Reagent, dyestuff, FITC or Dylight549 label
Sheep anti mouse fluorescence secondary antibody.
As a preferred embodiment of the invention, also recorded in the operational manual of the kit:
。
As a preferred embodiment of the invention, steps are as follows for the use of the kit:
(1) the postoperative tumor tissue section's white tiles of gastrointestinal stromal tumor patient is extracted, it is spare；
(2) white to the slice being prepared in step (1) respectively using Rad51 and KU70 protein immunization group detection reagent
Piece carries out Rad51 albumen and KU70 protein immunization histochemical staining, and to pathological section read tablet, according to immunohistochemistry calculation method meter
Calculate result；
(3) Rad51 is carried out using Rad51 and KU70 protein immunization group detection reagent difference normal tissue slice white tiles
Albumen and KU70 protein immunization histochemical staining, and to pathological section read tablet, according to immunohistochemistry calculation method calculated result；
(4) tumor tissues and normal tissue Rad51 albumen and KU70 albumen score ratio are calculated separately；
(5) above-mentioned calculating calculated result is determined to the drug resistance of tissue to be detected according to operational manual contents.
As a preferred embodiment of the invention, the ImmunohistochemistryMethods Methods include the following steps:
(1) dewaxing and aquation；
(2) it is washed using PBS；
(3) antigen retrieval；
(4) 3%H2O2Washing is washed with PBS；
(5) primary antibody is added to be incubated overnight；
(6) PBS is washed；
(7) secondary antibody is added；
(8) PBS is washed；
(9) the diluted horseradish enzyme label strepto- avidin of proper proportion is added dropwise to be incubated for；
(10) PBS is washed；
(11) chromogenic reagent；
(12) it rinses, mounting.
As a preferred embodiment of the invention, the medicine refers to the targeted drug of gastrointestinal stromal tumor, including her horse
For Buddhist nun and Sutent.
To realize above-mentioned second purpose, the technical solution adopted by the present invention is that:
Application of the kit as described above in prediction gastrointestinal stromal tumor drug resistance.
As a preferred embodiment of the invention, the medicine refers to the targeted drug of gastrointestinal stromal tumor, including her horse
For Buddhist nun and Sutent.
The chemical name of the Sutent is: (Z)-N- [2- (diethylin) ethyl -5- [(fluoro- 2- oxo -1,2- of 5-
Dihydro -3H- indoles -3- subunit) methyl] -2,4- dimethyl -3- carbamyl -1H- pyrroles's malate, molecular formula is:
C22H27FN4O2·C4H6O5, indication is as follows:
(1) imatinib mesylate treatment failure or intolerable gastrointestinal stromal tumor (GIST)；
(2) inoperable advanced renal cell carcinoma (RCC)；
(3) unresectable, metastatic differentiated progressive stage pancreas neuroendocrine tumors (pNET) adult patients.
It is 50mg that it, which treats gastrointestinal stromal tumor and the recommended dose of advanced renal cell carcinoma, once a day, is taken orally；Medication 4 weeks,
It is discontinued 2 weeks.
For pancreas neuroendocrine tumors, recommended dose 37.5mg is taken orally, once a day, continuous to take medicine, no drug withdrawal
Phase.
It is for gastrointestinal stromal tumor and metastatic renal cell cancer, according to the safety and tolerance of individual patients, with
12.5mg is that gradient units gradually adjust dosage.Daily maximum dose level is no more than 75mg, lowest dose level 25mg.
For pancreas neuroendocrine tumors, according to the safety and tolerance of individual patients, using 12.5mg as gradient units
Gradually adjust dosage.The maximum dose used in 3 clinical trial phases is daily 50mg.
As a preferred embodiment of the invention, the structural formula of the Imatinib is as follows:
As a preferred embodiment of the invention, the chemical name of the Imatinib is: 4- [(4- methyl-1-piperazine
Piperazine base) methyl]-N- [4- methyl -3- [4- (3- pyridyl group) -2- pyrimidine radicals] amino] phenyl] benzamide；Its molecular formula is:
C29H31N7O。
The invention has the advantages that:
1, kit of the invention can be used for predicting gastrointestinal stromal tumor drug resistance, thus pointedly selection targeting medicine
Object is greatly saved treatment time for gastrointestinal stromal tumor patient, provides to the treatment of gastrointestinal stromal tumor patient very big
It helps.
2, kit forecasting efficiency of the invention is high, can be applied in oncotherapy well, before having application well
Scape.
Specific embodiment
The invention will be further elucidated with reference to specific embodiments.It should be understood that these embodiments are merely to illustrate this hair
It is bright rather than limit the scope of the invention.In addition, it should also be understood that, after having read the content of the invention recorded, art technology
Personnel can make various changes or modifications the present invention, and such equivalent forms equally fall within the application the appended claims and limited
Fixed range.
Embodiment 1 predicts gastrointestinal stromal tumor drug resistance kit
It predicts to include operational manual in the kit of gastrointestinal stromal tumor drug resistance,
The operational manual records:
The operational manual also records:
The score computation method are as follows: staining power * stained area；
It further include that normal tissue sample, Rad51 protein immunization group detection reagent and KU70 albumen are exempted from the kit
Epidemic disease group detection reagent, the Rad51 protein immunization group detection reagent include rabbit-anti people Rad51 polyclonal antibody, Rad51 egg
The white goat antirabbit fluorescence secondary antibody for extracting reagent, dyestuff, FITC or Dylight549 label；The KU70 protein immunization groupization inspection
Test agent includes the mountain of the anti-human KU70 monoclonal antibody of mouse, KU70 Protein Extraction Reagent, dyestuff, FITC or Dylight549 label
Sheep anti mouse fluorescence secondary antibody.
The application method of the prediction gastrointestinal stromal tumor drug resistance kit of embodiment 2
Predict that the application method of the kit of patient tumors drug resistance is as follows:
(1) to the horizontal detection and calculating of patient's Rad51 protein expression:
(1) the postoperative tumor tissue section's white tiles of gastrointestinal stromal tumor patient is extracted according to the method for paraffin section de-waxing to water,
It is spare；
(2) the slice white tiles for taking step (1) to be prepared, distilled water flushing, and impregnated 5 minutes with PBS；
(3) antigen retrieval is carried out using the method for enzymic digestion；
(4) prepared 3%H is used2O2Incubation at room temperature 5-10 minutes, to eliminate the activity of endogenous peroxydase,
PBS is rinsed, and 2 minutes × 3 times；
(5) 5-10% normal rabbit serum is closed, and is incubated at room temperature 10 minutes.Incline serum deprivation, does not wash, and proper proportion dilution is added dropwise
Rabbit-anti people's primary antibody, 37 DEG C incubation 1-2 hours or 4 DEG C overnight；
(6) PBS is rinsed, and 2 minutes × 3 times；
(7) be added dropwise the diluted goat antirabbit secondary antibody of proper proportion (1%BSA-PBS dilution), 37 DEG C incubation 10-30 minutes；
37 DEG C or incubation at room temperature 10-20 minutes；
(8) PBS is rinsed, and 2 minutes × 3 times；
(9) diluted horseradish enzyme label strepto- avidin (PBS dilution) of proper proportion is added dropwise, 37 DEG C are incubated for 10~30 points
Clock, 37 DEG C or incubation at room temperature 10-20 minutes；
(10) PBS is rinsed, and 2 minutes × 3 times；
(11) chromogenic reagent；
(12) tap water sufficiently rinses, and redyes, and is dehydrated transparent, mounting；
(13) to pathological section read tablet according to immunohistochemistry calculation method calculated result, calculation method are as follows: staining power * dye
Color area；
(2) to the horizontal detection and calculating of patient's KU70 protein expression:
(1) it is white that the postoperative tumor tissue section of gastrointestinal stromal tumor patient is extracted according to the method for paraffin section de-waxing to water
Piece, it is spare；
(2) the above-mentioned slice white tiles being prepared, distilled water flushing are taken, and is impregnated 5 minutes with PBS；
(3) antigen retrieval is carried out using the method for enzymic digestion；
(4) prepared 3%H is used2O2Incubation at room temperature 5-10 minutes, to eliminate the activity of endogenous peroxydase,
PBS is rinsed, and 2 minutes × 3 times；
(5) 5-10% normal rabbit serum is closed, and is incubated at room temperature 10 minutes.Incline serum deprivation, does not wash, and proper proportion dilution is added dropwise
The anti-human primary antibody of mouse, 37 DEG C incubation 1-2 hours or 4 DEG C overnight；
(6) PBS is rinsed, and 2 minutes × 3 times；
(7) be added dropwise the diluted mountain sheep anti mouse secondary antibody of proper proportion (1%BSA-PBS dilution), 37 DEG C incubation 10-30 minutes；
37 DEG C or incubation at room temperature 10-20 minutes；
(8) PBS is rinsed, and 2 minutes × 3 times；
(9) diluted horseradish enzyme label strepto- avidin (PBS dilution) of proper proportion is added dropwise, 37 DEG C are incubated for 10~30 points
Clock, 37 DEG C or incubation at room temperature 10-20 minutes；
(10) PBS is rinsed, and 2 minutes × 3 times；
(11) chromogenic reagent；
(12) tap water sufficiently rinses, and redyes, and is dehydrated transparent, mounting；
(13) to pathological section read tablet, according to immunohistochemistry calculation method calculated result, calculation method are as follows: staining power *
Stained area；
(3), the horizontal detection and calculating of normal tissue Rad51 albumen and KU70 protein expression:
(1) the normal tissue sections white tiles in kit, distilled water flushing are taken, and is impregnated 5 minutes with PBS；
(2) antigen retrieval is carried out using the method for enzymic digestion；
(3) prepared 3%H is used2O2Incubation at room temperature 5-10 minutes, to eliminate the activity of endogenous peroxydase,
PBS is rinsed, and 2 minutes × 3 times；
(4) 5-10% normal rabbit serum is closed, and is incubated at room temperature 10 minutes.Incline serum deprivation, does not wash, and proper proportion dilution is added dropwise
The anti-human primary antibody of mouse, 37 DEG C incubation 1-2 hours or 4 DEG C overnight；
(5) PBS is rinsed, and 2 minutes × 3 times；
(6) be added dropwise the diluted mountain sheep anti mouse secondary antibody of proper proportion (1%BSA-PBS dilution), 37 DEG C incubation 10-30 minutes；
37 DEG C or incubation at room temperature 10-20 minutes；
(7) PBS is rinsed, and 2 minutes × 3 times；
(8) diluted horseradish enzyme label strepto- avidin (PBS dilution) of proper proportion is added dropwise, 37 DEG C are incubated for 10~30 points
Clock, 37 DEG C or incubation at room temperature 10-20 minutes；
(9) PBS is rinsed, and 2 minutes × 3 times；
(10) chromogenic reagent；
(11) tap water sufficiently rinses, and redyes, and is dehydrated transparent, mounting；
(12) to pathological section read tablet, according to immunohistochemistry calculation method calculated result, calculation method are as follows: staining power *
Stained area；
(4) above-mentioned calculating calculated result is determined to the drug resistance of tissue to be detected according to operational manual contents.
3 clinical verification of embodiment
Experiment in the present embodiment is to carry out under the premise of my informed consent, and sign informed consent form.
1 data
1.1 general information
Collecting in May, -2018 in November, 2015, Jinshan Hospital Fudan University is diagnosed as gastrointestinal stromal tumor patient 180
Example.
1.2 diagnostic criteria
Shape of tumor: tumor size is different, from 0.2cm~44cm etc., originates from gastrointestinal tract wall muscularis propria, can be to chamber
Outside interior, chamber or simultaneously to intracavitary, chamber outgrowth.Ulcer can be formed to Intracavity, therefore can be divided into according to tumour body position
Gastrointestinal tract external form in intracavitary type, wall inner mold, dumbbell shape, chamber external form and abdomen.Most of tumours are in expansion growth, clear border, matter
It is hard frangible；Section flesh of fish shape, bois de rose, center can have bleeding, necrosis, cystis degeneration etc. are secondary to sexually revise.Tumor number can be more
It is a.
Histological characteristic: GISTs is mainly made of spindle cell and epithelioid cell, and two kinds of cells can come across simultaneously
In different tumours, but morphological change range is big.According to two kinds of cells number can be divided into spindle cell type, epithelioid cell
Type and shuttle shape and epithelial cell mixed type.The arrangement of tumour cell is also in diversification, in the majority with pencil and sheet arrangement.Stomach with
The morphological change of small intestine is big, and the morphological change of rectum is small, and most of is spindle cell type, and it is more to intersect pencil arrangement.Tumour
Cell differentiation etc..
1.3 are included in standard
(1) meet above-mentioned gastrointestinal stromal tumor diagnostic criteria；
(2) age, men and women was unlimited between 18-60 years old；
(3) subject signs informed consent form.
1.4 reject standard:
The case for not meeting the standard of being included in and being accidentally included in；Though not controlled with this research approach after meeting the standard of being included in and being included in
The case for the treatment of；Non- curative effect reason and adverse reaction and test midway stop the case that falls off；Add with other drugs person；Data is not complete
It cannot the person of statistics.
1.5 terminate and remove clinical test standard: other diseases occur in test, influence to test into passerby；It can not be pre-
Other situations seen.
2 methods
2.1 experimental method
2.1.1 patient's biopsy tissues processing method
The postoperative tumor tissue section's white tiles of gastrointestinal stromal tumor patient is extracted according to the method for paraffin section de-waxing to water, it is standby
With；
According to kit in above-described embodiment 2 application method to the Rad51 albumen of the biopsy tissues of 180 patients with
KU70 albumen carries out immunohistochemical staining and calculates scores.
Tumor tissues and normal tissue Rad51, Ku70 protein expression level score are calculated according to immunohistochemistry calculation method,
And the ratio both calculated, content record its drug resistance knot for then recording calculated result according to kit operational manual
Fruit.
2.1.2 patient's post surgery treatment method
After 180 specimens slices are removed, gives patient's Imatinib Pharmaceutical and treat and observe, treat
Time is 6 months, and during treatment, whether close observation patient will appear drug resistance of tumor, and result is recorded after treatment time.
Usage and dosage: should be taken at table and drinks a mug water.Usually adult each 400mg or 600mg, Yi Ji once a day
Dose 800mg, that is, 400mg dosage, 2 times a day (in the morning and at night).The patient (including children) that tablet cannot be swallowed, can
To disperse tablet in the water or cider without gas (100mg agreement that contracts a film or TV play to an actor or actress 50ml, 400mg about use 200ml).It should stir mixed
Suspension, once tablet disintegration should take immediately completely.
2.2 standards of grading
It is carried out according to immunohistochemistry standards of grading
Immunohistochemical staining result distribute an average score, both considered a clear positive reaction staining power and
The ratio of tumour cell.
Positive reaction is defined as those cytoplasms and shows brown signal.For δ-catenin, a staining index
(0-12) is defined as the product of staining power and pigmented section.
The score of staining power is decided to be: 0 point negative；Weak 1 point；Medium 2 points；Strong positive 3 divides.The frequency quilt of positive cell
Is defined as: less than 5%, 0 point；5%-25%, 1 point；26%-50%, 2 points；51%-75%, 3 points；Greater than 75%, 4 point.
When dyeing isomery, we score in this way: each scoring is independent, the result is that comprehensive.
When data are analyzed, 0-7 is considered as low expression and 8 to 12 be considered as high expression.
3 results
Calculated result are as follows: 42 patients: (tumor tissues Rad51 score/normal tissue Rad51 score) > 1, (tumor tissues
Ku70 score/normal tissue Ku70 score) < 1, according to the operational manual of kit, determine that these patients are not in drug resistance
Property；42 patient's post-operative medications results are as follows: 41 patients do not occur drug resistance during treatment, and 1 patient occurs resistance to
Pharmacological property, it is 97.62% that kit operational manual, which verifies accuracy,.
46 patients: (tumor tissues Rad51 score/normal tissue Rad51 score) < 1, (tumor tissues Ku70 score/just
Often tissue Ku70 score) > 1, according to the operational manual of kit, determine that these patients will appear drug resistance in a short time；This 46
Example patient's post-operative medications result are as follows: drug resistance occur in a short time during treatment in 45 patients, and 1 patient does not occur drug resistance
Property, it is 97.82% that kit operational manual, which verifies accuracy,.
45 patients: (tumor tissues Rad51 score/normal tissue Rad51 score) > 1, (tumor tissues Ku70 score/just
Often tissue Ku70 score) > 1, according to the operational manual of kit, determine that these patients need close follow-up；45 patients
Post-operative medications result are as follows: the drug resistance of 43 patient's tumor tissues during treatment be it is uncertain as a result, it is desirable to closely with
It visits, occurs drug resistance during 2 patient's treatments in a short time, it is 95.56% that kit operational manual, which verifies accuracy,.
47 patients: (tumor tissues Rad51 score/normal tissue Rad51 score) < 1, (tumor tissues Ku70 score/just
Often tissue Ku70 score) < 1, according to the operational manual of kit, determine that these patients need close follow-up；47 patients
Post-operative medications result are as follows: the drug resistance of 45 patient's tumor tissues during treatment be it is uncertain as a result, it is desirable to closely with
It visits, occurs drug resistance during 2 patient's treatments in a short time, it is 95.74% that kit operational manual, which verifies accuracy,.
It should be noted that accuracy refers to that giving patient to the result of its drug resistance and kit during drug therapy grasps
Secretary's record drug resistance result is explained to calculate.
" drug resistance " refers to that disease is still in progress after medication or stable disease was less than 6 months；
" intolerance " refers to through stable disease in treatment 6 months, does not occur progression of disease.
In conclusion the accuracy of the operational manual of kit of the invention is 95.74% or more.
Kit of the invention is applied to prediction gastrointestinal stromal tumor patient tumors drug resistance, to facilitate under patient
The treatment of one step is greatly saved treatment time for patient, patient is helped to get well as early as possible.
Kit of the invention can be used for predicting gastrointestinal stromal tumor drug resistance, so that specific aim selects targeted drug, it is right
The treatment of gastrointestinal stromal tumor patient provides a great help.Kit forecasting efficiency of the invention is high, can apply well
In oncotherapy, have a good application prospect.
The above is only a preferred embodiment of the present invention, it is noted that for the ordinary skill people of the art
Member, without departing from the principle of the present invention, can also make several improvement and supplement, these are improved and supplement also should be regarded as
Protection scope of the present invention.
Claims (6)
1. a kind of kit for predicting gastrointestinal stromal tumor drug resistance, which is characterized in that the kit includes operational manual, institute
Operational manual is stated to record:
The score computation method are as follows: staining power * stained area；
It further include normal tissue sample, Rad51 protein immunization group detection reagent and KU70 protein immunization group in the kit
Change detection reagent, the Rad51 protein immunization group detection reagent includes that rabbit-anti people Rad51 polyclonal antibody, Rad51 albumen mention
Take the goat antirabbit fluorescence secondary antibody of reagent, dyestuff, FITC or Dylight549 label；The KU70 protein immunization groupization detection examination
Agent includes that the goat of the anti-human KU70 monoclonal antibody of mouse, KU70 Protein Extraction Reagent, dyestuff, FITC or Dylight549 label is anti-
Mouse fluorescence secondary antibody.
2. kit according to claim 1, which is characterized in that also recorded in the operational manual of the kit:
。
3. kit according to claim 1, which is characterized in that steps are as follows for the use of the kit:
(1) the postoperative tumor tissue section's white tiles of gastrointestinal stromal tumor patient is extracted, it is spare；
(2) using Rad51 and KU70 protein immunization group detection reagent respectively to the slice white tiles being prepared in step (1) into
Row Rad51 albumen and KU70 protein immunization histochemical staining, and to pathological section read tablet, it calculates and ties according to immunohistochemistry calculation method
Fruit；
(3) Rad51 albumen is carried out using Rad51 and KU70 protein immunization group detection reagent difference normal tissue slice white tiles
With KU70 protein immunization histochemical staining, and to pathological section read tablet, according to immunohistochemistry calculation method calculated result；
(4) tumor tissues and normal tissue Rad51 albumen and KU70 albumen score ratio are calculated separately；
(5) above-mentioned calculating calculated result is determined to the drug resistance of tissue to be detected according to operational manual contents.
4. kit according to claim 1, which is characterized in that the medicine refers to the targeted drug of gastrointestinal stromal tumor, including
Imatinib and Sutent.
5. application of the kit described in claim 1-4 in prediction gastrointestinal stromal tumor drug resistance.
6. applying according to claim 5, which is characterized in that the medicine refers to the targeted drug of gastrointestinal stromal tumor, including she
Imatinib and Sutent.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112048557A (en) * | 2020-09-01 | 2020-12-08 | 云南省肿瘤医院(昆明医科大学第三附属医院) | Kit for predicting gastrointestinal stromal tumor drug resistance and application thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2007120726A2 (en) * | 2006-04-11 | 2007-10-25 | The Regents Of The University Of California | Compositions and methods related to rad51 inactivation in the treatment of neoplastic diseases, and especially cml |
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| CN112048557A (en) * | 2020-09-01 | 2020-12-08 | 云南省肿瘤医院(昆明医科大学第三附属医院) | Kit for predicting gastrointestinal stromal tumor drug resistance and application thereof |
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