CN108977545B - A kind of ovarian cancer diagnosis or prognostic portfolio object and its application - Google Patents
A kind of ovarian cancer diagnosis or prognostic portfolio object and its application Download PDFInfo
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Abstract
The invention discloses a kind of ovarian cancer diagnosis and prognostic portfolio object and its applications.A kind of ovarian cancer diagnosis or prognostic portfolio object, are made of PDK1, PRKAA2 and PRKAB1.Ovarian cancer prognosis composition of the present invention is preparing the application in ovarian cancer diagnosis kit or the kit for predicting ovarian cancer prognosis situation.It is a kind of for predicting the kit of ovarian cancer prognosis situation, comprising detect PDK1, PRKAA2 and PRKAB1 expression quantity reagent.Joint-detection PDK1, PRKAA2 and PRKAB1 three can effectively judge ovarian cancer patients prognosis, and single any one index that detects can not effectively judge the prognosis situation of ovarian cancer patients.The present invention provides effective prognostic indicator for clinic.
Description
Technical field
The invention belongs to biological diagnosis field, it is related to a kind of ovarian cancer diagnosis and prognostic portfolio object and its application.
Background technique
Oophoroma is the highest tumour of lethality in gynecological tumor, and survival rate is lower than 30% within 5 years.
The disease incidence of ovarian neoplasm can suffer from ranking the 7th in tumour in women, and the ranking in feminine proses
Second, case fatality rate can suffer from position of being number five in tumour in all women, and rank the first in feminine proses
Position.2017, about 22500, U.S. women was newly diagnosed as oophoroma, wherein 14100 women die of oophoroma, oophoroma
Have become the primary cause of death of U.S.'s gynecological tumor.As society assigns female life and the mentally aggravation of pressure, add
Economy, environment etc. influence, the morbidity and mortality of oophoroma are in obvious ascendant trend.
Due to could generally occur the simultaneous phenomenon of ovarian neoplasm, most of ovarian epithelial carcinoma in late cases
Patient can just be diagnosed in the terminal stage of a disease, and clinical treatment faces the challenge.Existing clinical treatment advanced ovarian cancer is mainly subtracted using tumour
Art of going out cooperates chemicotherapy, but most of Patients with Advanced Ovarian Carcinoma can all recur.So far, subtract the postoperative tumour cell that goes out to subtract
The degree of going out is considered as most important Index for diagnosis factor, but subtracts the degree of going out and change with operative doctor clinical experience, cannot
Effectively help the prognosis of clinical judgment ovarian cancer patients.Therefore only rely on need the tumour of experience accumulation subtract go out art come sentence in time
The prognosis of disconnected ovarian cancer patients, so as to adjust therapeutic scheme, it is far from being enough for extending its life cycle.
Nineteen twenty, Nobel laureate Otto H.Warburg find for the first time tumour cell under aerobic conditions, sugared ferment
Solution is still remarkably reinforced, referred to as Warburg effect.The it is proposed of Warburg effect has caused all circles scholar to tumour cell energy
The research of metabolism makes glycolysis become hot spot in tumour cell research.Then the study found that active glycolysis metabolism is to dislike
The property significant biochemical characteristics of tumour cell.As tumour cell glycolysis metabolism increases constantly explaining for mechanism and function
It is bright, it is just being concerned by the strategy that targeting glycolysis metabolism approach treats malignant tumour.
Pyruvic dehydrogenase kinase (PDK) is the negative growth factor of mitochondrial pyruvate acidohydrogenase complex (PDC), is passed through
The activity of PDC is adjusted to regulate and control the decarboxylic reaction of pyruvic acid.Key as connection glycolytic pathway and oxidative phosphorylation approach
Branch node plays highly important regulating and controlling effect in aerobic glycolysis metabolism.In recent years, its unconventionality expression and regulation exist
Effect during tumor development is taken seriously further.
PDK is a kind of and the homologous silk of protokaryon protein kinase/tyrosine protein kinase family, with eucaryote silk/junket ammonia
Gluconokinase sequence is homologous.There are four types of isodynamic enzymes by PDK: PDK1, PDK2, PDK3 and PDK4.They are predominantly located in mitochondrial matrix,
Sequence homology is up to 70%, and the difference of sequence is mainly in N-terminal.PDK1 is mainly expressed in heart；PDK2 is in various tissues
Wide expression and expression；PDK3 is mainly expressed in testis；PDK4 is mainly expressed in heart and skeletal muscle.Pyruvic dehydrogenase has 3
A phosphorylation site, PDK2, PDK3 and PDK4 intelligence phosphorylation site 1 and 2, only PDK1 can be with 3 sites of phosphorylation.So
PDK1 can inhibit for a long time PDC by 3 sites of phosphorylation pyruvic dehydrogenase, so that limiting pyruvic acid enters line grain
Body carries out oxidative phosphorylation, and carries out glycolysis in endochylema.Under normal circumstances, PDK is in holddown, and PDC is for a long time by acetone
Acidohydrogenase activation, catalysis pyruvic acid carry out tricarboxylic acid cycle and generate a large amount of ATP.And under certain pathological conditions, PDK constantly quilt
Activation, leads to the change of metabolic pathway in cell, participates in the vicious transformation of cell.With deeply being influenced with PET for basic research
System has been detected unconventionality expression in lung cancer, nasopharyngeal carcinoma, colon cancer in clinical extensive use, PDK1.But it is in women
Expression in genital system is on the knees of the gods, and especially whether the highest oophoroma of the death rate, PDK1 participate in oophoroma
Occurrence and development and its to clinical importance whether need to be explored and studied.
PDK adjusts pyruvic dehydrogenase, to adjust glycolysis and oxidative phosphorylation, its object is to generate a large amount of ATP
It is used for cellular uptake energy.And the first receptor of intracellular energy is the protein kinase (AMPK) of adenosine acid activation.
Adenylate cyclase (AMPK) is widely present in eucaryotic organism, is highly conserved in evolutionary process
Serine/threonine protein kitase (STK), major regulatory intracellular metabolism and energy balance.AMPK is by the Asia α, β and γ
The heterotrimer of unit composition, each subunit can be divided into different hypotypes, respectively α again1、α2、β1、β2、γ1、γ2With
γ3, different respectively by 7 kinds of genotypic expressions of PRKAA1, PRKAA2, PRKAB1, PRKAB2, PRKAG1, PRKAG2, PRKAG3
Subunit can combine to form a variety of different AMPK tripolymers with free arrangement.This research early period discovery, PRKAA2 and
PRKAB1 is expressed in oophoroma to be increased, and difference is statistically significant, remaining 5 kinds of genotype is benign swollen in oophoroma and ovary
No significant difference is expressed in tumor.AMPK is intracorporal energy regulator, iuntercellular pressure change, glucose sugar exhausts, anoxic and
Intracellular AMP/ATP ratio increases when ischaemic, can activate AMPK with the 172nd threonine of phosphorylation α subunit.
And under pathological state, such as tumour, metabolic syndrome, often inhibit with energetic supersession disorder and AMPK activation, therefore, AMPK
It is considered as treating the latent effect target spot of metabolic disease and tumour.
In recent years, the continuous intensification with people to tumour cell metabolism understanding, work of the AMPK in tumor development
With also receiving significant attention.Largely studies have shown that AMPK, which can pass through, adjusts the proliferation that cell metabolism inhibits tumour cell.It is swollen
The energetic supersession mode of oncocyte aerobic glycolysis is different from normal cell.Tumour cell is by increasing the intake of glucose and adding
Fast glycolysis synthesizes ATP to meet metabolic demand.The product acetone acid of tumour cell glycolysis enters mitochondria further generation
The ability decline thanked, therefore, has scholar to think, if can inhibit the Warburg effect of tumour cell, restores the aerobic of mitochondria
Energetic supersession, or can reach the effect for inhibiting tumor cell proliferation.Moreover, adjusting and mitochondria of the AMPK to energetic supersession
Function it is inseparable, energy plants of the mitochondria as cell also play vital effect in health and disease.
Studies have shown that mitochondria can influence the activity of AMPK, while AMPK is adjusted mitochondria also by various aspects.To sum up,
AMPK is with a wide range of applications in treatment and prevention of tumour.
But have not yet to see the report that PDK1, PRKAA2 and PRKAB1 joint are used for ovarian cancer prognosis and diagnosis.
Summary of the invention
The object of the present invention is to provide a kind of ovarian cancer diagnosis and prognostic portfolio object.
It is a further object of the present invention to provide the applications of the composition.
It is yet another object of the invention to provide a kind of for predicting the kit of ovarian cancer prognosis situation.
The purpose of the present invention can be achieved through the following technical solutions:
A kind of ovarian cancer diagnosis or prognostic portfolio object, are made of PDK1, PRKAA2 and PRKAB1.
Ovarian cancer prognosis composition of the present invention is preparing ovarian cancer diagnosis kit or prediction ovarian cancer prognosis feelings
Application in the kit of condition.
The described application composition preferably of the present invention as detection target spot prepare ovarian cancer diagnosis kit or
Predict the application in the kit of ovarian cancer prognosis situation.
The reagent for detecting PDK1, PRKAA2 and PRKAB1 expression quantity is preparing ovarian cancer diagnosis kit or prediction oophoroma
Application in the kit of prognosis situation.
The preferred fluorescence quantitative PCR detection PDK1 of reagent of detection PDK1, PRKAA2 and PRKAB1 expression quantity,
The reagent of PRKAA2, PRKAB1 mRNA expression.
The fluorescence quantitative PCR detection PDK1, PRKAA2, PRKAB1 mRNA expression reagent further preferably include
Detect the primer of PDK1: SEQ ID NO.1/SEQ ID NO.2；Detect the primer of PRKAA2: SEQ ID NO.3/SEQ ID
NO.4；Detect the primer of PRKAB1: SEQ ID NO.5/SEQ ID NO.6.
It is a kind of for predicting the kit of ovarian cancer prognosis situation, include detection PDK1, PRKAA2 and PRKAB1 expression quantity
Reagent.
The kit preferably includes the reagent of fluorescence quantitative PCR detection PDK1, PRKAA2, PRKAB1 mRNA expression.
The kit further preferably includes following primer: detecting the primer of PDK1: SEQ ID NO.1/SEQ ID
NO.2；Detect the primer of PRKAA2: SEQ ID NO.3/SEQ ID NO.4；Detect the primer of PRKAB1: SEQ ID NO.5/
SEQ ID NO.6。
The kit still more preferably further includes the primer for detecting β-actin: SEQ ID NO.7/SEQ ID
NO.8。
The utility model has the advantages that
The present invention provides effective prognostic indicator, joint-detection PDK1, PRKAA2 for oophoroma clinical treatment and diagnosis
And PRKAB1 three can effectively judge ovarian cancer patients prognosis, and single any one index of detection can not be effective
The prognosis situation of segment ovarian cancer patients；The risk of patient's poor prognosis of high expression PDK1, PRKAA2 and PRKAB1 simultaneously
Increase.Therefore, the reagent especially Primer composition for detecting above three index can be in preparation ovarian cancer patients prognostic agent
It is applied in box.
Joint-detection PDK1, AMPK α 2 and 1 three of AMPK β can increase the positive rate of ovarian cancer patients, therefore,
PDK1, PRKAA2 and PRKAB1 three can be used as the auxiliary diagnosis that detection target is used for oophoroma.
Detailed description of the invention
The expression of PDK1 mRNA in Fig. 1 ovarian cancer cell line (SK-OV-3) and control cell lines (HFL)
The expression of PDK1 mRNA in Fig. 2 ovarian cancer tissue and benign tumor of ovary tissue
The expression of PRKAA2 mRNA in Fig. 3 ovarian cancer cell line (SK-OV-3) and control cell lines (HFL)
The expression of PRKAA2 mRNA in Fig. 4 ovarian cancer tissue and benign tumor of ovary tissue
The expression of PRKAB1 mRNA in Fig. 5 ovarian cancer cell line (SK-OV-3) and control cell lines (HFL)
The expression of PRKAB1 mRNA in Fig. 6 ovarian cancer tissue and benign tumor of ovary tissue
Expression (immunohistochemical staining, the amplification factor 200 of PDK1, PRKAA2, PRKAB1 in Fig. 7 ovarian cancer tissue
×)
Caption: A to D is respectively 0 point (-), 4 points (+), 8 points (++), 12 points (+++)；A, B is judged to feminine gender, and C and D are judged to sun
Property.
Fig. 8 Kaplan-Meier analyzes the OC survival of different PDK1 expressions
Caption: 0=PDK1 low expression；1=PDK1 high expression
Fig. 9 Kaplan-Meier analyzes the OC survival of different PRKAA2 expressions
Caption: 0=PRKAA2 low expression；1=PRKAA2 high expression
Figure 10 Kaplan-Meier analyzes the OC survival of different PRKAB1 expressions
Caption: 0=PRKAB1 low expression；1=PRKAB1 high expression
Figure 11 Kaplan-Meier analyzes the OC patient of different expressions after PDK1, PRKAA2, PRKAB1 triple combination
Survival rate
Caption: low expression after 0=triple combination；High expression after 1=triple combination
Specific embodiment
Experimental subjects and main agents involved in following embodiment are as follows:
1. experimental subjects
This research is the tissue mark that patient is obtained under the approval of No.1 Attached Hospital, Nanjing Medical Univ Ethics Committee
Sheet and clinical data information.
17 oophoroma tumors are tissue-derived in January, 2013 in December, 2016 Jiangsu Prov. Tumour Hospital and Jiangsu Province people
People hospital gynecological surgery Operated Specimens.15 benign tumor of ovary Specimen origins are cured in January, 2015 to Nanjing in December, 2017
Attached Jiangning hospital, university, section and Jiangsu Prov. People's Hospital gynecological surgery Operated Specimens.The preoperative non-Radiotherapy chemotherapy of all cases or
Other adjuvant treatments, all cases are through definitive pathological diagnosis.
Ovarian cancer tissue's pathological section sample totally 93, wherein 27 ovarian cancer tissue's pathological section Specimen origins are in 2008
Year January to Nanjing Women and Children Healthcare Hospital's in December, 2016 performs the operation Operated Specimens, 22 ovarian cancer tissue's pathological section Specimen origins
In January, 2013 in December, 2016 Jiangsu Prov. Tumour Hospital's gynecological surgery Operated Specimens, 44 ovarian cancer tissue's pathological sections
Specimen origin is in January, 2011 in December, 2017 Nanjing drum tower hospital gynecological surgery Operated Specimens.93 oophoroma samples
Histological type include serous carcinoma 73, mucus cancer 20；Organizational hierarchy: in it is low differentiation 75, differentiated 18；FIGO
By stages: the I-II phase 37, the III-IV phase 56；Having lymphatic metastasis is 40.Benign tumor of ovary sample totally 46, including 26
Example Mucinous cystoadenoma and 20 serous cystadenomas, it is attached in December, 2017 Nanjing Medical University to derive from January, 2015
Belong to Jiangning hospital.The preoperative non-Radiotherapy chemotherapy of all cases or other adjuvant treatments, all cases are through definitive pathological diagnosis.
Cell strain used in this research is human oophoroma cell line SK-OV-3, is purchased from Shanghai Cell Bank of the Chinese Academy of Sciences.
2. main agents
1.1 reagent of table and manufacturer's title
3 main solutions and preparation of reagents
(1) 5.6%NaHCO3 5.6g NaHCO3+100ml ddH2O sufficiently dissolves, autoclave sterilization, 4 DEG C after cooling
It saves.PH is adjusted to use.
(2) Hanks liquid stoste A: 160g NaCl+2g MgSO is taken4.7H2O+8g KCl+2g MgCl2.6H2O is dissolved in
1L ddH2O, 4 degree of preservations.Stoste B: 3.04g Na is taken2HPO4.12H2O+1.2g KH2PO4+ 20.0g glucose, is dissolved in
800ml ddH2O；It is mixed with 0.4% phenol red solution of 100ml is now matched, 4 DEG C of preservations.Working solution: 50ml stoste A, 50ml stoste B,
900ml ddH2O is sub-packed in sterile glass vials after mixing, first one layer of masking foil adds one layer of brown paper sealing again, and high temperature and pressure is gone out
Bacterium 15min.After cooling, 4 DEG C of preservations, with preceding sterile 5.6%NaHCO3Adjust PH to 7.2-7.6.
(3) twin antibiotic solution respectively takes 1,000,000 U penicillin and streptomysin, is dissolved in the autoclaved ddH of 100ml2O, filter
It is dispensed after device filtering, -20 DEG C of preservations.
(4) McCoy ' the S 5A cell complete culture solution 10ml FBS of the twin antibiotic containing 10%FBS+1%, with 90ml
After McCoy ' S5A mixing, 5.6%NaHCO3It is adjusted to PH 7.2-7.6,4 DEG C of preservations.
(5) cells frozen storing liquid dimethyl sulfoxide (DMSO) and the culture solution containing 10%FBS press 1 ︰, 9 prepared before use.
(6)
(7) 0.1%DEPC water 1ml DEPC water+1L ddH20
Embodiment 1
PDK1, PRKAA2, PRKAB1 mRNA high are expressed in ovarian cancer cell line (SK-OV-3) and ovarian cancer tissue
In order to inquire into biological action of the PDK1 in ovarian cancer cell, we are detected by quantitative real-time PCR
The mRNA expression of ovarian cancer cell line SK-OV-3 intracellular PDK1, PRKAA2, PRKAB1, and made pair of HFL cell line
According to.
In order in pre-test body ovarian cancer cell whether with external ovarian cancer cell PDK1, PRKAA2, PRKAB1 expression
Whether consistent, we have collected 17 ovarian cancer tissues and 15 benign tumor of ovary tissues, and pass through real-time fluorescence quantitative PCR
Method carries out the detection of mRNA to this two groups of tissues.
(1) reverse transcription reaction:
20 μ l systems: RNA (is adjusted mixed to 100 μ g of μ g~500) and 4 μ 5 × PrimeScript of l RT Master Mix
It is even, and with RNase-free water polishing to 20 μ l of total volume；
Reverse transcription condition is 37 DEG C of 15min, 85 DEG C of 5s and 4 DEG C of ∞；- 20 DEG C save backup.
(2) real-time fluorescence quantitative PCR (Real-time PCR):
PCR primer: using the first chain of cDNA as template, using β-actin as internal reference, expand PDK1, PRKAA2 and
PRKAB1.Use ddH2Primer is dissolved to final concentration of 100 μm of ol/L by O, and -20 DEG C save backup, and working concentration is 10 μm of ol/
ml.Primer sequence is shown in Table 1.
1 PCR primer sequence list of table
Reaction system (20 μ l): 2 × SYBR Premix Dimer Eraser, 10 μ l, upstream primer (10 μm of ol/ml) and
Each 0.6 μ l of downstream primer (10 μm of ol/ml), ddH26 μ l and cDNA template of O, 2 μ l.
Amplification program (two-step method): 95 DEG C of initial denaturation 30s, 95 DEG C of denaturation 5s, 57 DEG C of annealing 30s, 72 DEG C of extension 34s, 52
A circulation.
(3) analysis of experimental results: β-actin makees internal reference, using relative quantification method.In tissue samples, each sample purpose
The Ct value that the Ct value of gene subtracts corresponding β-actin obtains the two difference, i.e. △ Ct=CtTarget gene-Ctβ-actin, each sample is opposite
Quantification of 2- △ Ct=2(Ct target gene-Ct β-actin), two groups of samples compare using non-paired t test or rank sum test.In cell line
In, correspond to negative control group, the expression multiple of target gene, which changes, uses 2-ΔΔC tMethod (Δ Δ Ct=Δ CtTarget gene-Δ
CtNC), two groups of samples compare using paired t-test.If three wells, test replication 3 times every time.
As shown in Fig. 1,3,5, the expression phase of the intracellular PDK1 of ovarian cancer cell SK-OV-3, PRKAA2, PRKAB1 mRNA
Obviously increase compared with HFL cell, difference is statistically significant (P < 0.05).As shown in Fig. 2,4,6, PDK1 in ovarian cancer tissue,
The expression of PRKAA2, PRKAB1 mRNA are apparently higher than benign tumor of ovary tissue, and difference have statistical significance (P <
0.05)。
As shown in figure 4, the expression of the intracellular PRKAA2 mRNA of ovarian cancer cell SK-OV-3 is compared HFL cell and is obviously increased
Height, difference are statistically significant (P < 0.05).
Embodiment 2
2.1 immunohistochemistry
(1) sample slice toasts 40 minutes in 80 DEG C of baking ovens.
(2) sample slice dewaxes twice through dimethylbenzene, each 13min；Graded ethanol (100%, 100%, 90%, 80%,
70%) aquation, each 5min, pure water embathe twice, each 5min；
(3) high pressure antigen retrieval: immunohistochemistry antigen retrieval buffer (the powder-type lemon for being diluted to working solution in right amount is taken
Lemon acid system) (1500ml) in pressure cooker, pressure cooker is placed on electromagnetic oven, it is high-power to be heated to boiling.Power is adjusted to
Moderate (500-1000W) will be sliced as on high temperature resistant staining rack, be put into the reparation liquid to have boiled, cover pot cover, buckle pressure
Power valve continues to heat.Timing 3 minutes when pressure valve starts jet.Then pressure cooker is left into electromagnetic oven, it is naturally cold at room temperature
But after ten minutes, accelerated to repair liquid cooling but in pot in pressure outer wall of wall shower with flowing water, slice, flowing water are taken out after being cooled to room temperature
Rear 0.01M PBS is rinsed well to rinse 3 times, every time 4 minutes.
(4) tissue surrounding liquid is dried, 50 μ l of normal sheep serum working solution is added dropwise, room temperature in wet box is placed in and closes 2h；
(5) 1:100 diluted monoclonal antibody PDK1, PRKAA2, PRKAB1 (ABcam public affairs are added dropwise after siphoning away confining liquid
Department), it is placed in wet box and is put into 4 DEG C of refrigerators overnight incubations；
(6) next day, 0.01M PBS embathe slice 4 times, each 5min；PBS is sucked, enzyme is added dropwise and marks sheep anti-mouse igg secondary antibody
(stepping new), 37 DEG C of incubation 15min；
(7) 0.01M PBS embathes slice 4 times, each 5min, sucks PBS, every slice plus the DAB developing solution newly prepared
(step new) 100 μ l control developing time under mirror, terminate (about 3 points of halfs) in due course, pure water rinsing；
(8) haematoxylin redyes 1min, and 1% hydrochloride alcohol breaks up 5s, and flowing water returns indigo plant, and gradient alcohol dehydration, dimethylbenzene is transparent,
Neutral gum mounting.
The judgement of 2.2 results
PDK1, PRKAA2, PRKAB1 dyeing are positioned at endochylema, occasionally in after birth and karyon, show as in shallow palm fibre to palm fibre
Coloured particles.It replaces primary antibody to do negative control with PBS, uses known positive section as positive control.According to staining cell percentage
Rate and dye levels are evaluated and are analyzed.It is observed and is sliced using blind, first randomly select 10 under low power lens (100 times of mirrors)
Region containing tumour cell, then 200 tumour cells are counted under high power lens (200 times of mirrors), calculate positive percentage.It is positive
Cell percentage scoring criteria: positive percentage < 5% is 0 point, and (5-25) % is 1 point, and (26-50) % is 2 points, (51-
75) % is 3 points, and > 75% is 4 points；Staining power scoring criteria: dye-free, light yellow, brown color, sepia be denoted as 0 respectively,
1,2,3 points.As a result react percentage according to positive tumor cell and dye levels scoring product carries out Comprehensive Assessment: 0-2 points are
(-), 3-4 points are (+), and 5-8 points are (++), and 9-12 points are (+++).Tissue staining conditions are observed with Olympus microscope and are clapped
According to.
Immunohistochemistry staining method detect PDK1, PRKAA2, PRKAB1 protein expression difference and with clinical stages, leaching
The parameters such as transfer, histological type are fawned on to compare using χ2It examines.Survival analysis is carried out using Kaplan-Meier method, and is passed through
Log-Rank Test is compared.It is statistically significant with P < 0.05.
Expression of 2.3 PDK1, PRKAA2, the PRKAB1 in oophoroma and benign tumor of ovary histotomy
PDK1, PRKAA2, PRKAB1 positive immunostaining substance in ovarian cancer tissue are in faint yellow to sepia
It is granular, it is distributed mainly in cytoplasm, after birth and karyon are rare (Fig. 7).
Differential expression of 2.4 PDK1 in oophoroma and benign tumor of ovary
Judgment criteria is considered as feminine gender with "-" and "+", and " ++ " and " +++ " is considered as the positive, then has 54.8% in ovarian cancer tissue
(51/93) positive expression of visible PDK1, and the positive expression in benign tumor of ovary group is 15.2% (7/46), the two
Statistically significant (the χ of PDK1 differential expression2=19.87, P < 0.001) (table 2).
Expression of 2 PDK1 of table in oophoroma and benign tumor of ovary
Differential expression of 2.5 PRKAA2 in oophoroma and benign tumor of ovary
Judgment criteria is considered as feminine gender with "-" and "+", and " ++ " and " +++ " is considered as the positive, then has 61.7% in ovarian cancer tissue
(29/47) positive expression of visible PRKAA2, and the positive expression in benign tumor of ovary group is 10.5% (2/19), the two
Statistically significant (the χ of differential expression2=14.01, P < 0.0001) (table 3).
Expression of 3 PRKAA2 of table in oophoroma and benign tumor of ovary
Differential expression of 2.6 PRKAB1 in oophoroma and benign tumor of ovary
Judgment criteria is considered as feminine gender with "-" and "+", and " ++ " and " +++ " is considered as the positive, then has 74.5% in ovarian cancer tissue
(35/47) positive expression of visible PRKAA2, and the positive expression in benign tumor of ovary group is 38.9% (7/19), the two
Statistically significant (the χ of differential expression2=7.21, P < 0.01) (table 4).
Expression of 4 PRKAB1 of table in oophoroma and benign tumor of ovary
Expression after 2.7 PDK1, PRKAA2, PRKAB1 triple combinations in oophoroma and benign tumor of ovary
Judgment criteria is judged to feminine gender with the full yin of three, has one among three for the positive, that is, is judged to the positive, then oophoroma
There is its visible positive expression of 97.9% (46/47) in tissue, and the positive expression in benign tumor of ovary group is 47.1% (8/
17), the statistically significant (χ of the differential expression of the two2=24.45, P < 0.0001) (table 5).
Expression of 5 PDK1, PRKAA2, PRKAB1 triple combination of table in oophoroma and benign tumor of ovary
2.8PDK1 and expression of PDK1, PRKAA2, PRKAB1 triple combination in oophoroma and benign tumor of ovary
PDK1 is detected respectively oophoroma and benign tumor of ovary positive rate and triple combination after detect oophoroma and ovum
The carcinoid positive rate of nest is compared respectively, there is the positive rate of oophoroma and benign tumor of ovary after analyzing triple combination
It significantly improves, especially with the obvious (χ of oophoroma2=27.17, P < 0.0001), difference has statistical significance.As a result such as table 6.
6 PDK1 of table and expression ratio of PDK1, PRKAA2, PRKAB1 triple combination in oophoroma and benign tumor of ovary
Compared with
Note: △ PDK1 positive rate vs. triple combination positive rate (oophoroma)
* PDK1 positive rate vs. triple combination positive rate (benign tumor of ovary)
2.9 PRKAA2 and expression of PDK1, PRKAA2, PRKAB1 triple combination in oophoroma and benign tumor of ovary
Compare
PRKAA2 is detected respectively oophoroma and benign tumor of ovary positive rate and triple combination after detect oophoroma and
The positive rate of benign tumor of ovary is compared respectively, and the positive rate of oophoroma and benign tumor of ovary is equal after analyzing triple combination
It is significantly increased, especially with the obvious (χ of oophoroma2=19.06, P < 0.0001), difference has statistical significance.As a result such as table 7.
7 PRKAA2 of table and expression of PDK1, PRKAA2, PRKAB1 triple combination in oophoroma and benign tumor of ovary
Compare
Note: △ PRKAA2 positive rate vs. triple combination positive rate (oophoroma)
* PRKAA2 positive rate vs. triple combination positive rate (benign tumor of ovary)
2.10 PRKAB1 and expression of PDK1, PRKAA2, PRKAB1 triple combination in oophoroma and benign tumor of ovary
Compare
PRKAB1 is detected respectively oophoroma and benign tumor of ovary positive rate and triple combination after detect oophoroma and
The positive rate of benign tumor of ovary is compared respectively, and the positive rate for analyzing oophoroma after triple combination is significantly increased (χ2=
10.84, P < 0.001), difference has statistical significance.And benign tumor of ovary positive rate is also improved, but difference does not have
Statistical significance.As a result such as table 8.
8 PRKAB1 of table and expression of PDK1, PRKAA2, PRKAB1 triple combination in oophoroma and benign tumor of ovary
Compare
Note: △ PRKAB1 positive rate vs. triple combination positive rate (oophoroma)
* PRKAB1 positive rate vs. triple combination positive rate (benign tumor of ovary)
Relationship between the expression and oophoroma clinical pathologic characteristic of 2.11 PDK1
PDK1 expression intensity and patient age, tumor differentiation degree, FIGO by stages, lymphatic metastasis, unilateral or bilateral
Whether morbidity and coating are complete without significant correlation (table 9).
Relationship between the expression and oophoroma clinical pathologic characteristic of 9 PDK1 of table
Relationship between the expression and oophoroma clinical pathologic characteristic of 2.12 PRKAA2
PRKAA2 expression intensity and patient age, tumor size, tumor differentiation degree, FIGO by stages, lymphatic metastasis,
Whether unilateral or bilateral morbidity and coating are complete without significant correlation (table 10).
Relationship between the expression and oophoroma clinical pathologic characteristic of 10 PRKAA2 of table
Relationship between the expression and oophoroma clinical pathologic characteristic of 2.13 PRKAB1
PRKAB1 expression intensity and patient age, tumor size, tumor differentiation degree, FIGO by stages, lymphatic metastasis,
Whether unilateral or bilateral morbidity and coating are complete without significant correlation (table 11).
Relationship between the expression and oophoroma clinical pathologic characteristic of 11 PRKAB1 of table
The relationship between expression and oophoroma clinical pathologic characteristic after 2.14 PDK1, PRKAA2, PRKAB1 triple combinations
Expression and oophoroma Clinical symptoms such as tumor differentiation degree, FIGO after PDK1, PRKAA2, PRKAB1 triple combination
By stages, this four lymphatic metastasis, the morbidity of single bilateral and coating integrality aspects are in significant related (P < 0.05), but with year
Age is not significantly related to.As a result such as table 12.
The recall rate of oophoroma is not only substantially increased after triple combination, and to its clinical various features in significant related.
Imply that triple combination's index can provide help on the Index for diagnosis of oophoroma.
Expression after 12 PDK1, PRKAA2, PRKAB1 triple combination of table and the relationship between oophoroma clinical pathologic characteristic
Relationship between embodiment 3 PDK1, PRKAA2 and the existence of PRKAB1 high low expression and ovarian cancer patients
Relationship between 3.1PDK1 high low expression and ovarian cancer patients existence
It requires to be grouped patient according to following grouping, and counts the survival of patients time, similarly hereinafter
Grouping requires as follows:
PDK1 low expression group: immunohistochemical method detection PDK1 is expressed as the case of (-) and (+)
PDK1 high expression group: immunohistochemical method detection PDK1 is expressed as the case of (++) and (+++)
As a result such as Fig. 8, P=0.195, PDK1 low expression group and PDK1 high expression group nothing in ovarian cancer patients life span
Notable difference.It can be seen that PDK1 high low expression is not significantly related to ovarian cancer patients survival region.
Relationship between 3.2 PRKAA2 high low expressions and ovarian cancer patients existence
Grouping requires as follows:
PRKAA2 low expression group: immunohistochemical method detection PRKAA2 is expressed as the case of (-) and (+)
PRKAA2 high expression group: immunohistochemical method detection PRKAA2 is expressed as the case of (++) and (+++)
As a result such as Fig. 9, P=0.137, PRKAA2 low expression group and PRKAA2 high expression group are in ovarian cancer patients life span
Upper no significant difference.It can be seen that PRKAA2 high low expression is not significantly related to ovarian cancer patients survival region.
Relationship between 3.3 PRKAB1 high low expressions and ovarian cancer patients existence
Grouping requires as follows:
PRKAB1 low expression group: immunohistochemical method detection PRKAB1 is expressed as the case of (-) and (+)
PRKAB1 high expression group: immunohistochemical method detection PRKAB1 is expressed as the case of (++) and (+++)
As a result if Figure 10, P=0.831, PRKAB1 low expression group and PRKAB1 high expression group are when ovarian cancer patients are survived
Between upper no significant difference.It can be seen that PRKAB1 high low expression is not significantly related to ovarian cancer patients survival region.
Relationship between high low expression after 3.4 PDK1, PRKAA2, PRKAB1 triple combinations and ovarian cancer patients existence
Grouping requires as follows:
Triple combination's low expression group: immunohistochemical method testing result is the case of (-), (+) and (++).
Triple combination's height expression group: immunohistochemical method testing result is the case of (+++).
As a result such as Figure 11, P=0.013, after triple combination, high low expression and the ovarian cancer patients existence of Joint Index are pre-
There is obvious correlation afterwards, and high expression group patient prognosis is bad compared with low expression group, difference is statistically significant.
Sequence table
<110>Wang Fang
<120>a kind of ovarian cancer diagnosis or prognostic portfolio object and its application
<160> 8
<170> SIPOSequenceListing 1.0
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Claims (5)
1. the composition being made of pyruvic dehydrogenase kinase 1, PRKAA2 and PRKAB1 predicts ovarian cancer prognosis situation in preparation
Kit in application.
2. application according to claim 1, it is characterised in that the composition predicts ovum in preparation as detection target spot
Application in the kit of nest cancer prognosis situation.
3. detecting pyruvic dehydrogenase kinase 1, the reagent of PRKAA2 and PRKAB1 expression quantity predicts ovarian cancer prognosis feelings in preparation
Application in the kit of condition.
4. application according to claim 3, it is characterised in that the detection pyruvic dehydrogenase kinase 1, PRKAA2 and
The reagent of PRKAB1 expression quantity is fluorescence quantitative PCR detection pyruvic dehydrogenase kinase 1, PRKAA2, PRKAB1 mRNA expression
Reagent.
5. application according to claim 4, it is characterised in that the fluorescence quantitative PCR detection pyruvic dehydrogenase swashs
The reagent that enzyme 1, PRKAA2, PRKAB1 mRNA are expressed includes the primer for detecting pyruvic dehydrogenase kinase 1: SEQ ID NO.1/
SEQ ID NO.2；Detect the primer of PRKAA2: SEQ ID NO.3/ SEQ ID NO.4；Detect the primer of PRKAB1: SEQ ID
NO.5/ SEQ ID NO.6。
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106053813A (en) * | 2016-06-25 | 2016-10-26 | 林森 | Immunohistochemical kit for detecting PDK1 expression quantity of nasopharyngeal carcinoma, and application of immunohistochemical kit |
| CN107050017A (en) * | 2017-03-29 | 2017-08-18 | 吉林大学 | PDK1 inhibitor, the composition containing the inhibitor and purposes |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106053813A (en) * | 2016-06-25 | 2016-10-26 | 林森 | Immunohistochemical kit for detecting PDK1 expression quantity of nasopharyngeal carcinoma, and application of immunohistochemical kit |
| CN107050017A (en) * | 2017-03-29 | 2017-08-18 | 吉林大学 | PDK1 inhibitor, the composition containing the inhibitor and purposes |
Non-Patent Citations (2)
| Title |
|---|
| An immunohistochemical perspective of PPARβ and one of its putative targets PDK1 in normal ovaries, benign and malignant ovarian tumours;N Ahmed等;《British Journal of Cancer》;20081231;第98卷;第1415-1424页 |
| Over-expressions of AMPK subunits in ovarian carcinomas with significant clinical implications;Cuilan Li等;《BMC Cancer》;20121231;第12卷;第1-8页 |
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