CN105497900A - Anti-undifferentiated-thyroid-carcinoma drug-resistant target spot and application thereof - Google Patents
Anti-undifferentiated-thyroid-carcinoma drug-resistant target spot and application thereof Download PDFInfo
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- CN105497900A CN105497900A CN201510216684.3A CN201510216684A CN105497900A CN 105497900 A CN105497900 A CN 105497900A CN 201510216684 A CN201510216684 A CN 201510216684A CN 105497900 A CN105497900 A CN 105497900A
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Abstract
The invention discloses an anti-undifferentiated-thyroid-carcinoma drug-resistant target spot and application thereof. The invention discloses influence of micro RNA-30d on undifferentiated thyroid drug treatment, and thus provides a purpose of a micro RNA-30d stimulant in drug for reducing or eliminating the undifferentiated thyroid carcinoma drug resistance and application of the micro RNA-30d stimulant as a undifferentiated thyroid carcinoma drug-resistant mark and as a drug-resistant tumor prediction and diagnosis and drug-resistant tumor drug design and screening target spot. The invention provides a novel mechanism for overcoming the tumor drug resistance and improving the curative effect, and provides a novel target spot for the anti-tumor drug resistance.
Description
Technical field
The invention belongs to molecular biology and treatment and prevention of tumour field, more specifically, the present invention relates to prediction, Diagnosis and Treat drug resistance of tumor field.
Background technology
Undifferentiated thyroid carcinoma is the one that in thyroid carcinoma, grade malignancy is the highest, and sickness rate accounts for the 10-15% of whole thyroid carcinoma, and such tumor growth is fast, can occur in early days to infiltrate and whole body transfer, and grade malignancy is high, poor prognosis.Undifferentiated thyroid carcinoma has the feature being difficult to diagnose in advance usually, because it has toleration to known treatment means.Understand the molecular mechanism causing this kind of drug resistance of tumor, finally to develop the Focal point and difficult point that the new treatment of reversing drug resistance is current undifferentiated thyroid carcinoma study on prevention field.
In order to reverse undifferentiated thyroid carcinoma drug resistance, improve chemotherapeutic efficacy, the new drug of tumor drug resistance in the urgent need to studying the mechanism of tumor drug resistance generation, the key gene target spot of tumor drug resistance in several ways, and is treated in this area according to these shot designs, thus guides tumor drug resistance magnetic target therapy.
The endogenous non-coding strand microRNA that the tetracycline-regulated gene of miRNAs to be a class length be 18 ~ 24 nucleotide is expressed, by the 3 ' untranslated region with said target mrna i.e. 3 ' UTR specific binding, said target mrna is caused to degrade or Translational repression, produce PTGS, current research proves, miRNA molecule has regulated and controled the gene expression of in human genome about 1/3.Large quantity research has confirmed that miRNAs plays a significant role, comprising the propagation, differentiation, apoptosis, Infiltration and metastasis etc. of regulating cell in the developing of malignant tumor.MiR-30d is positioned human chromosome 8q24.22, in malignant tumor, developing effect occurs be subject to extensive concern as the important a member in miR-30 family.Research report miR-30d up-regulated in the Several Kinds of Malignancies such as carcinoma of prostate, medulloblastoma, hepatocarcinoma, breast carcinoma.But miR-30d but presents low expression status in undifferentiated thyroid carcinoma, and participate in undifferentiated thyroid carcinoma malignant progression process.
Summary of the invention
The object of this part is some aspects of general introduction embodiments of the invention and briefly introduces some preferred embodiments.May do in the specification digest and denomination of invention of this part and the application a little simplify or omit with avoid making this part, specification digest and denomination of invention object fuzzy, and this simplification or omit and can not be used for limiting the scope of the invention.
In view of Problems existing in above-mentioned and/or existing tumour medicine and application thereof, propose the present invention.
Therefore, an object of the present invention is to provide a kind of key gene target spot and albumen target spot of tumor drug resistance, for reversing tumor drug resistance, raising chemotherapeutic efficacy provide more efficiently instrument.
For solving the problems of the technologies described above, the invention provides following technical scheme: a kind of anti-undifferentiated thyroid carcinoma drug resistance target spot, described target spot is miR-30d and/or its autophagy approach participated in.
Another object of the present invention is to provide a kind of miR-30d simulant and is reducing and/or eliminating the application in the medicine of the drug resistance in oncotherapy.
As miR-30d simulant of the present invention at a kind of preferred version reducing and/or eliminate the application in the medicine of the drug resistance in oncotherapy, wherein: described miR-30d simulant comprises: miR-30d expresses the autophagy approach restrainer of molecule, miR-30d mediation.
As miR-30d simulant of the present invention at a kind of preferred version reducing and/or eliminate the application in the medicine of the drug resistance in oncotherapy, wherein:
Described miR-30d expresses molecule, and its base sequence of expressing is GUUGUUGUAMCAUCCCCGACUGGMGCUGUMGACACAGCUMGCUUUCAGUCAGAUGU UUGCUGCUAC.
An also object of the present invention is to provide a kind of pharmaceutical composition, and it comprises: the carrier adopting miR-30d simulant, tumor therapeutic agent and pharmaceutically can accept.
As a kind of preferred version of pharmaceutical composition of the present invention, wherein: described tumor therapeutic agent comprises, one or more in cisplatin, amycin, paclitaxel, carboplatin etc., combretastatin A-4, Erlotinib, manumycin or Sorafenib.
As a kind of preferred version of pharmaceutical composition of the present invention, wherein: the described carrier that pharmaceutically can accept comprises: one or more in saline, buffer, glucose, water, glycerol or ethanol.
Another object of the present invention is to provide a kind of method of screening the anti-tumor medicine of overriding resistance, and it comprises: provide undifferentiated thyroid carcinoma tumor cell line, tumor culture or tumor animal; Drug candidate is contacted with the tumor cell line provided, tumor culture or tumor animal, as administration group; Detect the expression of miR-30d in administration group, and compare with the miR-30d expression not giving the control tumor cell line of drug candidate, tumor culture or tumor animal, if testing result shows, the expression of administration group miR-30d is significantly higher than matched group, then show that this drug candidate is the anti-tumor medicine of overriding resistance.
Beneficial effect of the present invention:
(I) relation between miR-30d and undifferentiated thyroid carcinoma drug resistance is disclosed, for design and screening drug-resistant tumor drug provide new approach and target spot;
(2) provide and improve the new method of undifferentiated thyroid carcinoma to the sensitivity of oncotherapy by raising miR-30d and related signaling pathway thereof, thus serve the positive role improving oncotherapy effect, reduce tumor therapeutic agent consumption, reduction of patient misery, there is clinical practicality.
Detailed description of the invention
For enabling above-mentioned purpose of the present invention, feature and advantage become apparent more, are described in detail the specific embodiment of the present invention below in conjunction with detailed description of the invention.
Set forth a lot of detail in the following description so that fully understand the present invention, but the present invention can also adopt other to be different from alternate manner described here to implement, those skilled in the art can when without prejudice to doing similar popularization when intension of the present invention, therefore the present invention is by the restriction of following public specific embodiment.
Secondly, alleged herein " embodiment " or " embodiment " refers to special characteristic, structure or the characteristic that can be contained at least one implementation of the present invention.Different local in this manual " in one embodiment " occurred not all refers to same embodiment, neither be independent or optionally mutually exclusive with other embodiments embodiment.
The present invention studies discovery: miR-30d is low expression in undifferentiated thyroid carcinoma tissue, and confirms that miR-30d is the key molecule of undifferentiated thyroid carcinoma generation chemoresistance by In vitroandin vivotrial.The miR-30d of low expression preferentially can start a series of anti-apoptotic bioprocess, and inhibited apoptosis, causes tumor drug resistance.Adopt simulant to raise miR-30d to express, can reduce or eliminate the drug resistance of tumor.On this basis, the present inventor completes the present invention.
One of main purpose of the present invention is exactly key gene target spot and albumen target spot in order to provide a kind of undifferentiated thyroid carcinoma drug resistance, and for reversing undifferentiated thyroid carcinoma drug resistance, raising chemotherapeutic efficacy provides more efficiently instrument.
Another main purpose of the present invention is according to the prediction of these shot designs, diagnosing tumour drug resistance test kit, screens the medicine of the treatment tumor drug resistance for this tiny RNA and correlation molecule signal path thereof.
An object of the present invention is to provide a kind of crucial target spot mir-30d reversing undifferentiated thyroid carcinoma drug resistance, for this target spot and associated signal paths thereof, as autophagy activation inhibitor, design medicine and therapeutic scheme, raise mir-30d in undifferentiated thyroid carcinoma express or suppress its function, effectively can treat drug-resistant tumor.
In an embodiment of the invention, described mir-30d such as, from people or other eukaryote, mice, rat, cattle or monkey etc., and has the conservative of height between them.
In an embodiment of the invention, described simulant is selected from the synthesis analogies for miR-30d.
In yet another embodiment of the present invention, described simulant is the expression plasmid for miR-30d, and the sequence of more preferably described expression is:
GUUGUUGUAMCAUCCCCGACUGGMGCUGUMGACACAGCUMGCUUUCAGUCAGAUGUUUGCUGCUAC。
In yet another embodiment of the present invention, described tumor is undifferentiated thyroid carcinoma.
In yet another embodiment of the present invention, the therapeutic agent that described oncotherapy adopts is selected from: alkylating agent, antimetabolite, antitumor antibiotics, plant anticarcinogen or immune formulation.
In a preference, described therapeutic agent is selected from: cisplatin, amycin, paclitaxel, carboplatin etc., combretastatin A-4, Erlotinib, manumycin, Sorafenib.
In another preference, described therapeutic agent is preferably platinum medicine: cisplatin or carboplatin.
In a second aspect of the present invention, provide a kind of pharmaceutical composition, described pharmaceutical composition comprises:
(i) miR-30d simulant;
(ii) tumor therapeutic agent;
(iii) pharmaceutically acceptable carrier or excipient.
In a preference, described tumor therapeutic agent is chemotherapeutics.
In a third aspect of the present invention, provide a kind of medicine box, described medicine box comprises:
(A) container of miR-30d simulant is accommodated;
(B) container and (C) operation instructions of tumor therapeutic agent are held.
In a preference, the simulant comprised in pharmaceutical composition of the present invention or medicine box is selected from: for expression molecule, the autophagy inhibitor chloroquine of miR-30d.
In another preference, described in pharmaceutical composition of the present invention or medicine box, simulant is the expression molecule for miR-30d, and more preferably described expressed sequence is:
GUUGUUGUAMCAUCCCCGACUGGMGCUGUMGACACAGCUMGCUUUCAGUCAGAUGUUUGCUGCUAC。
In another preference, described autophagy inhibitor chloroquine acts on the low autophagy activation signal path of expressing mediation of miR-30d.
In another preference, described tumor is selected from undifferentiated thyroid carcinoma.
In another preference, described therapeutic agent is selected from: cisplatin, amycin, paclitaxel, carboplatin etc., combretastatin A-4, Erlotinib, manumycin, Sorafenib.
In a fourth aspect of the present invention, provide a kind of method of screening the undifferentiated thyroid carcinoma medicine of overriding resistance, described method comprises:
A () provides undifferentiated thyroid carcinoma tumor cell line, tumor culture or tumor animal;
B drug candidate contacts with the tumor cell line provided in step (a), tumor culture or tumor animal, as administration group by ();
C () detects the expression of miR-30d in administration group, and compare with the expression of the miR-30d not giving the control tumor cell line of drug candidate, tumor culture or tumor animal;
If testing result shows, the expression of the miR-30d of administration group is significantly higher than matched group, then show that this drug candidate is the anti-tumor medicine of overriding resistance.
In a preference, described method can not produce the anti-tumor medicine of oncotherapy drug resistance to the individuality suffering from tumor for screening.
Disclosing miR-30d in the present invention is first a kind of undifferentiated thyroid carcinoma drug resistance-associated proteins, and it is abnormal low expression in tumor tissues, and a series of anti-apoptotic proteins process of preferential startup, inhibited apoptosis, causes tumor drug resistance.
Based on above-mentioned discovery of the present invention, those of ordinary skill in the art can using miR-30d or its participate in autophagy approach as target spot, design or screening overriding resistance tumour medicine.Such as, those of ordinary skill in the art according to known miR-30d sequential design or can filter out miR-30d simulant, jointly give tumor patient by itself and chemotherapeutics, thus improve tumor to the consumption of the drug resistance of medicine, the success rate improving oncotherapy, minimizing anti-tumor medicine and the misery alleviating patient.
MiR-30d simulant includes but not limited to: miR-30d expresses the autophagy approach restrainer of molecule, miR-30d mediation.Such as, in the embodiment of the present invention provide for miR-30d express base sequence: GUUGUUGUAMCAUCCCCGACUGGMGCUGUMGACACAGCUMGCUUUCAGUCAGAUGU UUGCUGCUAC.
With regard to miRNAs simulant, along with going deep into of miRNAs research, existing company commercialization can provide design and the Composite service of miRNAs
Obviously, by above-mentioned commercial sources and service, this area research worker only need provide target-gene sequence, just can obtain high-quality miR-30d simulant.
In addition, those of ordinary skill in the art also can adopt method as known in the art and software to come designed, designed and screening miRNA simulant.
Therefore, as long as those of ordinary skill in the art have known the sequence of target gene miR-30d, they just can obtain expression molecule for this tiny RNA by commercial sources easily, also can according to principle as known in the art, adopt known software and method to obtain expression molecule for this tiny RNA.
Pharmaceutical composition and medicine box
The invention provides a kind of pharmaceutical composition, it contains the described miR-30d analogies of effective dose, and pharmaceutically acceptable carrier.Pharmaceutical composition of the present invention can be used for, by expressing miR-30d or being combined with autophagy inhibitor, improving undifferentiated thyroid carcinoma drug resistance, improves the effectiveness of anti-tumor medicine in oncotherapy.
Acceptable carrier on pharmaceutical composition Chinese materia medica of the present invention, comprises (but being not limited to): saline, buffer, glucose, water, glycerol, ethanol and combination thereof.Usual pharmaceutical preparation should match with administering mode, and pharmaceutical composition of the present invention can be made into injection form, such as, be prepared by conventional method with normal saline or the aqueous solution containing glucose and other adjuvant.Described pharmaceutical composition should aseptically manufacture.The dosage of active component is treatment effective dose.Pharmaceutical preparation of the present invention also can be made into slow releasing preparation.
In pharmaceutical composition of the present invention, the effective dose of inhibitor can change with order of severity of the pattern of administration and disease to be treated etc.The selection of preferred effective dose can be determined (such as passing through clinical trial) according to various factors by those of ordinary skill in the art.Described factor includes but not limited to: inhibitor of the present invention with the use of the kind of tumor therapeutic agent and consumption; Pharmacokinetic parameter such as bioavailability, metabolism, the half-life etc. of inhibitor; The order of severity of the disease that patient will treat, the body weight of patient, the immune state of patient, the approach etc. of administration.Such as, by an urgent demand for the treatment of situation, the dosage that several times separate can be given every day, or dosage is reduced pari passu.
The administering mode of pharmaceutical composition of the present invention has no particular limits, and can be whole body or local.Preferably, pharmaceutical composition of the present invention gives object by intravenous mode.Also directly simulant can be delivered medicine to experimenter by the method such as such as injecting; Or, miR-30d ceneme (such as expression vector or virus etc.) will be carried by certain approach and be delivered on target spot, and make it expression activity miR-30d.
Anti-tumor medicine and simulant of the present invention can be formulated in same pharmaceutical composition, or be provided in the form of individual in medicine box.Medicine and simulant of the present invention can produce synergy when conbined usage, and the conbined usage of such as miR-30d and cisplatin can be treated undifferentiated thyroid carcinoma and be produced synergism.Those of ordinary skill in the art can be combined into row filter according to the conventional means (such as with reference to the method disclosed in the embodiment of the present invention) in the present invention to these.
The method of the anti-tumor medicine of screening overriding resistance
Additionally providing a kind of in the present invention is target spot with miR-30d, the method of the undifferentiated thyroid carcinoma medicine (such as anti-tumor medicine) of screening overriding resistance, described method comprises: (a) provides undifferentiated thyroid carcinoma tumor cell line, tumor culture or tumor animal; B drug candidate contacts with the tumor cell line provided in step (a), tumor culture or tumor animal, as administration group by (); C () detects the expression of miR-30d in administration group, and compare with the miR-30d expression not giving the control tumor cell line of drug candidate, tumor culture or tumor animal; If testing result shows, the expression of administration group miR-30d is significantly higher than matched group, then show that this drug candidate is the anti-tumor medicine of overriding resistance.
Method of the present invention can be used for from compound library, filter out the non-drug resistance antitumor drug for tumor, can be used as an important indicator of screening compound assessment.Method of the present invention also can be used for clinical in in the prediction of the personalized treatment of tumor patient and diagnosis, filter out the antitumor drug for individuality with best oncotherapy effect.
As can be seen here, a kind of new tumor drug resistance mark is provided: low expression in undifferentiated thyroid carcinoma tissue also causes the microRNA-30d of tumor drug resistance in the present invention, using this tiny RNA as target spot, can design for the prediction of this tiny RNA and correlation molecule thereof, diagnosing tumour drug resistance test kit, screen the medicine of the treatment undifferentiated thyroid carcinoma drug resistance for this tiny RNA and correlation molecule thereof.
It should be noted that, above embodiment is only in order to illustrate technical scheme of the present invention and unrestricted, although with reference to preferred embodiment to invention has been detailed description, those of ordinary skill in the art is to be understood that, can modify to technical scheme of the present invention or equivalent replacement, and not departing from the spirit and scope of technical solution of the present invention, it all should be encompassed in the middle of right of the present invention.
Claims (8)
1. an anti-undifferentiated thyroid carcinoma drug resistance target spot, is characterized in that, described target spot is miR-30d and/or its autophagy approach participated in.
2. a miR-30d simulant is reducing and/or is eliminating the application in the medicine of the drug resistance in oncotherapy.
3. miR-30d simulant according to claim 2 is reducing and/or is eliminating the application in the medicine of the drug resistance in oncotherapy, it is characterized in that, described miR-30d simulant comprises: miR-30d expresses the autophagy approach restrainer of molecule, miR-30d mediation.
4. miR-30d simulant according to claim 3 is reducing and/or eliminating the application in the medicine of the drug resistance in oncotherapy, it is characterized in that, described miR-30d expresses molecule, and its base sequence of expressing is
GUUGUUGUAMCAUCCCCGACUGGMGCUGUMGACACAGCUMGCUUUCAGUCAGAUGUUUGCUGCUAC。
5. a pharmaceutical composition, is characterized in that, comprising: miR-30d simulant as claimed in claim 2, tumor therapeutic agent and the carrier that pharmaceutically can accept.
6. pharmaceutical composition according to claim 5, is characterized in that, described tumor therapeutic agent comprises, one or more in cisplatin, amycin, paclitaxel, carboplatin etc., combretastatin A-4, Erlotinib, manumycin or Sorafenib.
7. pharmaceutical composition according to claim 5, is characterized in that, the described carrier that pharmaceutically can accept comprises: one or more in saline, buffer, glucose, water, glycerol or ethanol.
8. screen a method for the anti-tumor medicine of overriding resistance, it is characterized in that, comprising:
Undifferentiated thyroid carcinoma tumor cell line, tumor culture or tumor animal are provided;
Drug candidate is contacted with the tumor cell line provided, tumor culture or tumor animal, as administration group;
Detect the expression of miR-30d in administration group, and compare with the miR-30d expression not giving the control tumor cell line of drug candidate, tumor culture or tumor animal, if testing result shows, the expression of administration group miR-30d is significantly higher than matched group, then show that this drug candidate is the anti-tumor medicine of overriding resistance.
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| CN107312778A (en) * | 2017-07-21 | 2017-11-03 | 申永智 | A kind of cancer diagnosing kit and medicine for treatment compositions |
| CN115400216A (en) * | 2022-10-08 | 2022-11-29 | 浙江省人民医院 | Pharmaceutical composition for thyroid undifferentiated carcinoma and application |
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Application publication date: 20160420 |