CN108913773A - The polymolecular marker and its device and evaluation method of a kind of clinical evaluation oophoroma platinum-based chemotherapy sensibility - Google Patents
The polymolecular marker and its device and evaluation method of a kind of clinical evaluation oophoroma platinum-based chemotherapy sensibility Download PDFInfo
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- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
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- C12Q2600/00—Oligonucleotides characterized by their use
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- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
Abstract
The polymolecular marker and its device and evaluation method of a kind of clinical evaluation oophoroma platinum-based chemotherapy sensibility, screen chemotherapy side effect gene simultaneously from multiple data sets, and the prediction effect of the polymolecular marker is verified and fully assessed in tens independent data sets, it ensure that the present invention in the potential using value of oophoroma clinic platinum-based chemotherapy sensitivity assessment, polymolecular marker of the present invention only includes 16 genes, it is easy to clinical test detection and reduces testing cost, the tumor tissues sample of patient is easily obtained, patient can be sampled by syringe needle and directly carry out determination of gene expression；The polymolecular marker not will receive the influence of the batch effect or detection platform difference of experiment；The polymolecular marker does not need to carry out before use the data normalization processing between multisample, easy to use.In the drug resistance of chemotherapy real-time monitoring patient, personalized therapeutic scheme is taken in time convenient for clinician, just find after avoiding patient from taking turns high toxicity medication resistant more.
Description
Technical field
The present invention relates to cma gene group and oncology technical field, in particular to a kind of clinical evaluation oophoroma
The polymolecular marker and its device and evaluation method of platinum-based chemotherapy sensibility.
Background technique
Oophoroma is the gynecologic cancers type of most lethal, at the same be also women in be number five position cancer cause
Cause of the death element.Because of the concealment of its onset, early stage non-evident sympton, diffusion is fast, recurs the features such as morning and the death rate are high, to the health of women
The threat got worse is constituted with life.Global to have more than 220,000 oophoroma new cases every year, death toll is up to 140,000.?
China, oophoroma are the first causes of death of gynecologic malignant tumor.According to Chinese ovarian tumors rates in 2011 and death rate tune
It looks into, newly-increased oophoroma case 4.5 ten thousand, the death rate annual 1.8 ten thousand are consistent with global incidence every year for China.Oophoroma early stage nothing
Manifest symptom, 70% or more has been advanced stage when finding, therefore it is only 40%-45% that 5 years survival rates are lower.Current main treatment
Means are to assist 6~8 course for the treatment of of paclitaxel plus platinum chemotherapy after surgical cytoreduction and operation.But due to ovum
Nest cancer patient is to the difference of chemotherapy side effect, in addition the heterogeneity of tumour itself, so that chemosensitivity is there is huge difference,
About 30% patient is just found to have chemoresistant after receiving more wheel adjuvant chemotherapy, has not only delayed the best opportunity for the treatment of
The huge toxicity of chemotherapy has been also subjected to it, the life cycle of serious curtailment patient and quality of life.And for remaining 70% disease
Even if people can generate reaction completely when initial to chemotherapy, but still have up to 75% patient to recur in several years.
Tumour cell is that oophoroma is difficult to obtain the master of definite curative effect to the drug resistance of the intrinsic either acquisition of platinum-based chemotherapy
Want obstacle.Therefore the platinum-based chemotherapy sensibility that patient how is precisely predicted before chemotherapy and after more wheel chemotherapy, reduces and uses
Medicine blindness instructs clinician to select personalized supplemental treatment regimens, extends evening for further increasing the curative effect of chemotherapy
Phase and the life cycle of patients with recurrent are a problems in the urgent need to address in the accurate medical domain of oophoroma.
Since different ovarian cancer patients are to the significant difference of platinum-based chemotherapy sensibility, only facing according to patient with doctor
Bed feature and clinical experience are difficult to the sensibility of accurate judgement patient's platinum-based chemotherapy, so that at present clinically frequently with platinum class
Drug is undesirable come the effect for carrying out chemotherapy.With the rapid development of molecular biology and cell biology, researcher and clinic
Doctor focus be transferred to how using biomarker go prediction platinum-based chemotherapy sensibility.Previous research stops more
In terms of single-gene, the gene pleiomorphism or in terms of metabolism group, it is albumen and genomic dna sequence that the research method of use is also mostly
Variation.As what the development of high throughput gene expression detection technique and transcript profile were studied deepens continuously, researcher has found gene
Important regulating and controlling effect is all played in transcription and post-transcriptional level, although unstable under alkaline condition, but still easily be detected and
Quantization.Different antibody, different interpretation methods are often used from protein labeling analyte detection, possible difference of testing result etc. lacks
Point is compared, and RNA marker is more sensitive, and specificity is stronger, and testing cost is lower.It is quasi- with genomic DNA sequence variations detection means
True property is relatively high to be interpreted relatively objective, and repeatability is strong etc. compares, and gene marker more dynamic reflects cell state and regulation
Process.
It is more that multiple groups research has been found that the heterogeneity of occurrence and development and the curative effect of medication reaction of oophoroma suffers from
The synergistic effect of genetic and environmental factor.Multiple high throughput expression analysis research discovery chemosensitivities and drug resistant breast cancer, ovum
Gene expression profile in the kinds of tumors tissue such as nest cancer is different, and explanation should be one group of gene, rather than the expression of a gene
Heterogeneity can influence the curative effect and bright sensibility of drug, and influence of the single-gene to chemosensitivity may be fallen into oblivion by many factors
Not yet, early period is unilateral to the prediction of chemosensitivity and evaluation comparison based on single-gene marker, and sensibility and specificity is often inadequate
It is ideal.Therefore by quick to the screening of chemosensitivity gene, chemotherapy using existing transcription group and Clinical symptoms big data
The identification of predisposing genes function and the optimal polymolecular marker group of selection merge corresponding quick using mathematical modeling principles foundation
Perceptual score in predicting model can provide more effective and accurate chemotherapy drug susceptibility evaluation index for ovarian cancer patients,
To facilitate clinicians make individualized treatment scheme, reduce medication blindness, chemotherapeutic efficacy and judging prognosis are monitored, is mentioned
The survival rate of high ovarian cancer patients.
Summary of the invention
In order to make up for the deficiencies of the prior art, the purpose of the present invention is to provide a kind of clinical evaluation oophoroma platinum medicines
The polymolecular marker and its device and evaluation method of chemosensitivity, polymolecular marker provided by the present invention combination for
Oophoroma platinum-based chemotherapy sensibility and prognosis prediction sensitivity and specificity with higher, can be used as novel molecular mark
Object is used to instruct the selection and optimization of one line of oophoroma and two wires personalized chemotherapeutic regimens.
The technical solution that the present invention uses is：More points of a kind of clinical evaluation oophoroma platinum-based chemotherapy sensibility
Sub- marker, the polymolecular marker include the drug susceptibility of up-regulated expression in drug susceptibility patient or cell line
The drug resistance gene marker of up-regulated expression, the drug are quick in gene marker and drug resistance patient or cell line
Predisposing genes marker includes 6 marker gene marker FZD4, MUTYH, PCK2, PEX10, SRPK1, UCP2, described
Drug resistance gene marker includes 10 markers gene marker EDIL3, GNG12, MBOAT2, MTMR6, NBR1, NEK7,
NET1, PPP3CA,RAD17,WDR41。
A kind of device using polymolecular marker clinical evaluation oophoroma platinum-based chemotherapy sensibility, the device
Evaluation model including calculating sensitivity scores, the formula of the evaluation model are as follows：
Wherein, T is test statistics,For the average value of 6 drug susceptibilty gene marker expression levels,For
The average value of 10 drug resistance gene marker expression levels, n1For the number of drug susceptibilty gene marker, n2For drug
The number of resistant gene marker,For the variance of drug susceptibilty gene marker expression level,For drug resistance gene
The variance of marker expression level.
A kind of oophoroma platinum class medicine of the polymolecular marker using clinical evaluation oophoroma platinum-based chemotherapy sensibility
The evaluation method of object chemosensitivity, includes the following steps：
(1) it obtains platinum-based chemotherapy treated oophoroma full-length genome and expresses data, the oophoroma full-length genome
Expressing data includes two sets of human ovarian cancer patients' microarray datas, a set of ovarian cancer cell line chip data and a set of ovary carninomatosis
People's two generations sequencing data；
(2) by the original chip of two sets of human ovarian cancer patients' microarray datas of acquisition and a set of ovarian cancer cell line chip
Data carry out data prediction and standardization using RMA algorithm；
(3) the expression data obtained after data processing are thin in the two class samples or two classes of medicaments insensitive and drug resistance
Difference expression gene is screened using limma and edgeR software package respectively between born of the same parents system；
(4) difference expression gene for obtaining step (3) is using the ranking of preceding each data set in preceding 1000 difference tables
Up to gene, at least 16 difference expression genes occurred will be concentrated more as one group of platinum-based chemotherapy sensibility in three data
Molecular marker is obtained including FZD4, MUTYH, PCK2, PEX10, SRPK1, UCP2, EDIL3, GNG12, MBOAT2,
The polymolecular marker of MTMR6, NBR1, NEK7, NET1, PPP3CA, RAD17, WDR41；
(5) 16 difference expression genes of acquisition are divided into 6 upper mileometer adjustments in drug susceptibility patient or cell line
Drug susceptibilty gene the marker FZD4, MUTYH, PCK2, PEX10, SRPK1, UCP2 reached；It is anti-in drug with other 10
Drug resistance gene marker EDIL3, GNG12, MBOAT2, MTMR6, the NBR1 of up-regulated expression in venereal disease people or cell line,
NEK7,NET1,PPP3CA,RAD17, WDR41；
(6) by the expression of the expression and 10 drug resistance gene markers of 6 drug susceptibilty gene markers
Level carries out double totality T and examines, and for the T test statistics that double totality T are examined as sensitivity scores, formula is as follows：
Wherein,It is quick for 6 drugs
The average value of predisposing genes marker expression level,For the average value of 10 drug resistance gene marker expression levels, n1
For the number of drug susceptibilty gene marker, n2For the number of drug resistance gene marker,For drug susceptibilty gene
The variance of marker expression level,For the variance of drug resistance gene marker expression level；
(7) T test statistics will be obtained to evaluate, if T<0, it is resistant to platinum-based chemotherapy to disclose the patient,
Chemotherapeutic efficacy is low；If T>0, disclosing the patient has sensibility to platinum-based chemotherapy, and chemotherapeutic efficacy is high.
The beneficial effects of the invention are as follows：The present invention provides a kind of clinical evaluation oophoroma platinum-based chemotherapy sensibility
Polymolecular marker and its device and evaluation method screen chemotherapy side effect gene simultaneously from multiple data sets, and to this more points
The prediction effect of sub- marker is verified and has been fully assessed in tens independent data sets, ensure that the present invention in ovary
The potential using value of cancer clinic platinum-based chemotherapy sensitivity assessment, polymolecular marker of the present invention only include 16 bases
Cause is easy to clinical test detection and reduces testing cost, and the tumor tissues sample of patient is easily obtained, and can pass through syringe needle pair
Patient's sampling directly carries out determination of gene expression；The polymolecular marker not will receive the batch effect or detection platform of experiment
The influence of difference；The polymolecular marker does not need to carry out before use the data normalization processing between multisample, easy to use.
In the drug resistance of chemotherapy real-time monitoring patient, takes personalized therapeutic scheme in time convenient for clinician, avoid patient
It is just found after more wheel high toxicity medications resistant.
Specific embodiment
It in order to illustrate more clearly of the content of present invention, is described as follows with specific embodiment, specific embodiment does not limit this hair
Bright context.
1) it obtains platinum-based chemotherapy treated oophoroma full-length genome and expresses data, the oophoroma full-length genome table
It include two sets of human ovarian cancer patients' microarray datas, a set of ovarian cancer cell line chip data and a set of human ovarian cancer patients up to data
Two generation sequencing datas.
2) by above-mentioned steps 1) obtain Affymetrix HG-U133A and HG-U133_Plus_2 platform two sets of ovaries
The original chip data of carninomatosis people and a set of ovarian cancer cell line chip carry out data prediction and standardization using RMA algorithm,
The expression data that other platform datas are directly handled well using author.
3) by above-mentioned steps 2) obtain four sets express data medicaments insensitive and drug resistance two class samples or two
Difference expression gene is screened using limma and edgeR software package respectively between class cell line.
4) by above-mentioned steps 3) obtain difference expression gene we only with preceding each data set ranking preceding 1000
A difference expression gene at least will concentrate 16 difference expression genes occurred as one group of platinum-based chemotherapy in three data
Sensibility polymolecular marker, the polymolecular marker include FZD4, MUTYH, PCK2, PEX10, SRPK1, UCP2, EDIL3,
GNG12,MBOAT2,MTMR6,NBR1,NEK7, NET1,PPP3CA,RAD17,WDR41。
5) above-mentioned steps 4) polymolecular marker in, 6 marker genes (FZD4, MUTYH, PCK2, PEX10,
SRPK1, UCP2) it is drug susceptibilty gene, their up-regulated expressions in drug susceptibility patient or cell line；And other 10
A marker gene (EDIL3, GNG12, MBOAT2, MTMR6, NBR1, NEK7, NET1, PPP3CA, RAD17, WDR41) is medicine
Object resistant gene, their up-regulated expressions in drug resistance patient or cell line.
6) we are by the table of the expression of 6 drug susceptibilty gene markers and 10 drug resistance gene markers
It carries out double totality T up to level to examine, for the T test statistics that double totality T are examined as sensitivity scores, formula is as follows：
Wherein,For the average value of 6 drug susceptibilty gene marker expression levels,For 10 drug resistance genes
The average value of marker expression level, n1For the number of drug susceptibilty gene marker, n2For drug resistance gene marker
Number,For the variance of drug susceptibilty gene marker expression level,Horizontal for drug resistance gene marker expression
Variance.
7) expression of this 16 polymolecular markers 6) is carried out integration foundation through the above steps can clinical application
Platinum-based chemotherapy sensitivity prediction scoring model, 0 be used as cut off value, patient is divided into drug susceptibility and drug resistance,
If T<0, disclose that the patient is resistant to platinum-based chemotherapy, and chemotherapeutic efficacy is low；If T>0, the patient is disclosed to platinum medicine
Chemotherapy has sensibility, and chemotherapeutic efficacy is high.
Present invention firstly discovers that a kind of new polymolecular marker based on 16 genes, which can be to ovum
Nest cancer platinum-based chemotherapy sensibility is evaluated and is able to carry out the Index for diagnosis of patient.Compared with the existing technology, this is more
Molecular marker is advantageous in that：
First, the present invention screens chemotherapy side effect gene simultaneously from multiple data sets, and to the pre- of the polymolecular marker
It surveys effect to be verified and fully assessed in tens independent data sets, the application of above method and strategy ensure that this
Potential using value of the invention in oophoroma clinic platinum-based chemotherapy sensitivity assessment；
Second, which only includes 16 genes, is easy to clinical test detection and reduces testing cost；
The tumor tissues sample of third, patient is easily obtained, and can be sampled by syringe needle to patient and directly be carried out gene table
Up to measurement；
4th, which not will receive the influence of the batch effect or detection platform difference of experiment；
5th, which does not need to carry out before use the data normalization processing between multisample, user
Just；
6th, in the drug resistance of chemotherapy real-time monitoring patient, take personalized treatment in time convenient for clinician
Scheme is just found resistant more after avoiding patient from taking turns high toxicity medication.
In the description of the present invention, it should be noted that unless otherwise clearly defined and limited, term " installation ", " phase
Even ", " connection " shall be understood in a broad sense, for example, it may be being fixedly connected, may be a detachable connection, or be integrally connected；It can
To be mechanical connection, it is also possible to be electrically connected；It can be directly connected, can also can be indirectly connected through an intermediary
Connection inside two elements.For the ordinary skill in the art, above-mentioned term can be understood at this with concrete condition
Concrete meaning in invention.In addition, in the description of the present invention, unless otherwise indicated, the meaning of " plurality " is two or two
More than.
Every technical staff's notice：Although the present invention is described according to above-mentioned specific embodiment, of the invention
Invention thought be not limited in the invention, any repacking with inventive concept will all be included in this patent protection of the patent right
In range.
The above is only a preferred embodiment of the present invention, protection scope of the present invention is not limited merely to above-mentioned implementation
Example, all technical solutions belonged under thinking of the present invention all belong to the scope of protection of the present invention.It should be pointed out that for the art
Those of ordinary skill for, several improvements and modifications without departing from the principles of the present invention, these improvements and modifications
It should be regarded as protection scope of the present invention.
Claims (3)
1. a kind of polymolecular marker of clinical evaluation oophoroma platinum-based chemotherapy sensibility, which is characterized in that described is more
Molecular marker includes the drug susceptibilty gene marker and drug of up-regulated expression in drug susceptibility patient or cell line
The drug resistance gene marker of up-regulated expression, the drug susceptibilty gene marker packet in resistance patient or cell line
Include 6 marker gene markers FZD4, MUTYH, PCK2, PEX10, SRPK1, UCP2, the drug resistance gene
Marker includes 10 markers gene marker EDIL3, GNG12, MBOAT2, MTMR6, NBR1, NEK7, NET1,
PPP3CA, RAD17, WDR41。
2. a kind of dress using polymolecular marker clinical evaluation oophoroma platinum-based chemotherapy sensibility described in claim 1
It sets, which is characterized in that the device includes the evaluation model for calculating sensitivity scores, and the formula of the evaluation model is such as
Under：
；
Wherein, T is test statistics,For the average value of 6 drug susceptibilty gene marker expression levels,For 10 medicines
The average value of object resistant gene marker expression level,For the number of drug susceptibilty gene marker,For drug resistance
The number of gene marker,For the variance of drug susceptibilty gene marker expression level,For drug resistance gene mark
The variance of object expression.
3. it is a kind of using right want 1 described in clinical evaluation oophoroma platinum-based chemotherapy sensibility polymolecular marker ovum
The evaluation method of nest cancer platinum-based chemotherapy sensibility, which is characterized in that include the following steps：
（1）It obtains platinum-based chemotherapy treated oophoroma full-length genome and expresses data, the oophoroma full-length genome expression
Data include two sets of human ovarian cancer patients' microarray datas, a set of ovarian cancer cell line chip data and a set of human ovarian cancer patients two
For sequencing data；
（2）By the original chip number of two sets of human ovarian cancer patients' microarray datas of acquisition and a set of ovarian cancer cell line chip
Data prediction and standardization are carried out according to using RMA algorithm；
（3）By the expression data obtained after data processing medicaments insensitive and drug resistance two class samples or two class cell lines
Between using respectively limma and edgeR software package screen difference expression gene；
（4）By step（3）The difference expression gene of acquisition is using the ranking of preceding each data set in preceding 1000 differential expression bases
Cause at least will concentrate 16 difference expression genes occurred as one group of platinum-based chemotherapy sensibility polymolecular in three data
Marker is obtained including FZD4, MUTYH, PCK2, PEX10, SRPK1, UCP2, EDIL3, GNG12, MBOAT2,
The polymolecular marker of MTMR6, NBR1, NEK7, NET1, PPP3CA, RAD17, WDR41；
（5）16 difference expression genes of acquisition are divided into 6 up-regulated expressions in drug susceptibility patient or cell line
Drug susceptibilty gene marker FZD4, MUTYH, PCK2, PEX10, SRPK1, UCP2；It is anti-in drug with other 10
Drug resistance gene marker EDIL3, GNG12, MBOAT2, the MTMR6 of up-regulated expression in venereal disease people or cell line,
NBR1, NEK7, NET1, PPP3CA, RAD17, WDR41；
（6）By the expression of the expression of 6 drug susceptibilty gene markers and 10 drug resistance gene markers
It carries out double totality T to examine, for the T test statistics that double totality T are examined as sensitivity scores, formula is as follows：
；Wherein,For 6 drug susceptibilty genes
The average value of marker expression level,For the average value of 10 drug resistance gene marker expression levels,It is quick for drug
The number of predisposing genes marker,For the number of drug resistance gene marker,For drug susceptibilty gene marker table
Up to horizontal variance,For the variance of drug resistance gene marker expression level；
（7）It will obtain T test statistics to evaluate, if T<0, it is resistant to platinum-based chemotherapy to disclose the patient, chemotherapy
Curative effect is low；If T>0, disclosing the patient has sensibility to platinum-based chemotherapy, and chemotherapeutic efficacy is high.
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| CN113025713A (en) * | 2021-02-23 | 2021-06-25 | 温州医科大学 | Use of biomarkers for predicting the sensitivity of a tumor patient to a specific anti-tumor drug |
| CN113025713B (en) * | 2021-02-23 | 2022-11-22 | 浙江东睿生物科技有限公司 | Use of biomarkers for predicting the sensitivity of a tumor patient to a specific anti-tumor drug |
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