CN106701767B - MicroRNA molecule miR-663a relevant to vincristine drug resistance and its application - Google Patents
MicroRNA molecule miR-663a relevant to vincristine drug resistance and its application Download PDFInfo
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Abstract
The present invention provides a kind of microRNA molecule miR-663a relevant to vincristine drug resistance and its applications, belong to genetic engineering and technical field of cancer biotherapy.The nucleotide sequence of miR-663a of the present invention is as shown in SEQ ID NO.1, it has high in vincristine mdr cell express, in the characteristic of vincristine sensitive cells strain low expression, it can be used as vincristine drug resistance marker, it is used to prepare for colon cancer vincristine drug resistance auxiliary diagnostic box, also provides new target spot for design colon cancer vincristine medicine-resistant medicine.The present invention provides application of the miRNA marker in terms of preparing the vincristine drug resistance auxiliary diagnostic box for treating colon cancer, screening new treatment of colon cancer drug, and the miRNA inhibitor is preparing the application in antagonism vincristine medicine-resistant medicine, has preferable clinical value.
Description
Technical field
The present invention relates to genetic engineerings and technical field of cancer biotherapy, relate in particular to a kind of resistance to vincristine
The relevant microRNA molecule miR-663a of pharmacological property and its application.
Background technique
Colorectal cancer is one of most common malignant tumor of digestive tract, and the disease incidence of colorectal cancer just increases year by year, hair
Sick rate is only second to the malignant tumours such as the cancer of the esophagus, gastric cancer and lung cancer.Vincristine (vincristine, VCR) is that have at present extensively
The antitumor chemotherapeutics used.However, long-term treatment can make tumour cell to VCR generate drug resistance, this is to cause
One of the main reason for chemotherapy fails.Data show that 90% cancer death is related with the drug resistance of tumour.
MicroRNA (miRNA) is a kind of endogenous, highly conserved non-coding microRNA with adjusting function, length
About 20~25 nucleotide.Have been found that it is related with many biological phenomenas, such as development, break up and apoptosis, tumour occur also lead
Relate to the exception of miRNA expression.In addition, also some researches show that miRNA with tumour cell to the sensitivity phase of chemicotherapy
It closes.MiRNA can wide participation growth cycle, cell Proliferation and differentiation, Apoptosis, metabolism, neuromodulation, tumour occur
And the regulation process of the various physiology and pathology such as interaction of virus and host.In cell genetic fragment copy number variation,
Insertion, missing and the Genome stabilities abnormal phenomenon such as numerical abnormalities of chromosomes can cause some miRNA unconventionality expression or
Regulation.Mutation, abnormal expression or the abnormal normal function that will affect miRNA of processing modification of miRNA, leads to the table of target gene
Up to changing, the drug susceptibility of tumour cell is influenced.
Vincristine plays an important role in the therapeutic process of colon cancer, but the drug resistance phenomenon occurred is to limit it to face
The main reason for bed curative effect.Specify biological function and mechanism of the new micoRNA in colon cancer cell vincristine drug resistance
New biological target is provided to effectively improve the clinical efficacy of colon cancer.
Summary of the invention
The purpose of the present invention is to provide a kind of microRNA molecule miR-663a relevant to vincristine drug resistance and
It is applied.
The present invention is passed through using high throughput sequencing technologies of new generation in conjunction with bioinformatic analysis method and experimental verification
Colon cancer vincristine drug-resistant cell strain is established, skill is sequenced with HiSeq 2500 in the research strategy screened using full-length genome
The method of art and bioinformatics, detection and analysis drug-resistant cell strain and non-drug resistant cell strain differential expression
MiRNAs has found miR-663a high expression in vincristine mdr cell.
One kind provided by the invention microRNA molecule relevant to vincristine drug resistance, is miR-663a, nucleosides
Acid sequence contains sequence shown in SEQ ID NO.1.
The present invention provides the biomaterial containing miR-663a, the biomaterial is carrier, host cell, transgenosis
Cell line, engineering bacteria.
The present invention provides miR-663a molecule or containing its biomaterial as colon cancer vincristine drug resistance mark
Application in will object.
The present invention provides miR-663a molecule or containing its biomaterial preparation colon cancer vincristine drug resistance it is auxiliary
Help the application in diagnostic kit.
In above-mentioned application, the microRNA molecule high expression in vincristine mdr cell.
The present invention provides miR-663a molecules to screen the application in new treatment of colon cancer drug.
The present invention provides the inhibitor of miR-663a molecule to prepare the application in the drug resistant drug of antagonism vincristine.
The present invention provides a kind of for treating the drug of colon cancer, contains miR-663a molecule inhibitor.
Specific primer for PCR detection miR-663a molecule is to belonging to the scope of protection of the present invention.
In an embodiment of the present invention, for detecting the specific primer of miR-663a molecule to for SEQ ID NO.2-4
It is shown.
The present invention provides above-mentioned specific primers in preparation colon cancer vincristine drug resistance auxiliary diagnostic box
Application.
The beneficial effects of the present invention are the nucleotide sequence of miR-663a of the present invention contains as shown in SEQ ID NO.1
Sequence, there is in vincristine mdr cell high expression, in the characteristic of vincristine sensitive cells strain low expression, can make
For vincristine drug resistance marker, it is used to prepare for colon cancer vincristine drug resistance auxiliary diagnostic box, it can be specifically
The expression quantity for detecting miR-663a in tissue, is efficiently used for the clinical drug resistant auxiliary diagnosis of colon cancer vincristine；The present invention
New target spot also is provided for design colon cancer vincristine medicine-resistant medicine, and on this basis, to design and develop miR-
The inhibitor of 663a reduces to the application in vincristine drug resistance drug in preparation or is screening new colon using it as target spot
Application in cancer therapeutic agent, precisely to be treated, improved curative effect, to reduce side effect.The present invention provides miRNA marks
The will object treatment of colon cancer new in vincristine drug resistance auxiliary diagnostic box and screening of the preparation for treating colon cancer
Application in terms of drug has preferable clinical value and wide application prospect.
Detailed description of the invention
Fig. 1 is that cell Proliferation-toxicity test detects VCR sensitive cells HCT-8 and drug-resistant cell strain HCT-8/VCR to Changchun
The drug resistance ability effect picture of new alkali.
Fig. 2 is the expression of miR-663a in VCR sensitive cells strain HCT-8 and drug-resistant cell strain HCT-8/VCR cell.
Fig. 3 is that cell Proliferation-toxicity test detects HCT-8 cell transfecting miR-663a analogies transfection VCR to the resistance to of VCR
Medicine ability effect picture.
Fig. 4 is that cell Proliferation-toxicity test detects HCT-8/VCR cell transfecting miR-663a inhibitor to vincristine
Drug resistance ability effect picture.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..Without departing substantially from spirit of that invention
In the case where essence, to modifications or substitutions made by the method for the present invention, step or condition, all belong to the scope of the present invention.
Unless otherwise specified, chemical reagent used in embodiment is conventional commercial reagent, skill used in embodiment
The conventional means that art means are well known to those skilled in the art.Cell line used, reagent etc. are in embodiment and test example
Commercial goods.HCT-8 cell is purchased from Shanghai Cell Bank of the Chinese Academy of Sciences.
Embodiment 1 constructs vincristine medicine-resistant cell line HCT-8/VCR
Colon cancer drug-resistant cell strain HCT-8/VCR is established using the method for being stepped up vincristine (VCR) concentration.It will be quick
Sense cell HCT-8 is incubated in the culture solution containing VCR, and initial concentration 5ng/mL gradually increases drug concentration, respectively later
10,100,1 000 and 2 000ng/mL.It is good thin with limiting dilution method clonal growth after each concentration obtains drug resistance
Born of the same parents, subsequently into the culture and screening of next concentration.Last HCT-8 cell persistently cultivated for 20 generations in the VCR of 2000ng/mL
More than, medicine-resistant cell line HCT-8/VCR is constructed, tests first 1 week and deactivates VCR.
2 cell of embodiment tests (mtt assay) to the drug sensitivity of VCR
The persister HCT-8/VCR and sensitive strain cell HCT-8 of logarithmic growth phase respectively is dilute by serum free medium
It is 1 × 10 that interpretation of the law tally, which counts cell density,5A/mL, and with 1 × 104The cell solution in a/hole is inoculated with the training of 96 hole cells
It supports in plate.The culture solution for being free of VCR is added in blank control group, and experimental group is separately added into containing 10ng/ml, 20ng/ml, 100ng/
The vincristine culture solution of ml, 200ng/ml, 1000ng/ml, 2000ng/ml, 3000ng/ml, 4000ng/ml, 100 holes μ l/.
Group of cells cultivates 48h in 37 DEG C of incubations；Culture plate is taken out, 20 hole μ l/ of MTT (5mg/mL) solution is added, is continued
It is put into 37 DEG C of cell incubators and incubates 4h；Then it is inhaled with pipettor and abandons culture supernatant, 150 hole μ l/ DMSO is added, sufficiently shakes
After swinging, culture plate is put into enzyme-linked immunosorbent assay instrument, sets Detection wavelength as 570nm, reads absorbance (OD) value in each hole.
Background cell mean is calculated, and is returned to zero using this average value as zero point, and calculate group of cells survival rate: is thin
Born of the same parents' survival rate=(experimental group mean OD value-background group mean OD value)/blank control group mean OD value-background group mean OD value)
× 100%, then calculate drug concentration when 50% cell survival, i.e. half-inhibitory concentration IC50.
Colon cancer cell HCT-8 and HCT-8/VCR are handled using the VCR of various concentration, Fig. 1 is shown in the inhibiting rate of cell.
The 503nhibiting concentration IC50 of HCT-8 cell and HCT-8/VCR are respectively 140.265 and 2350.469.
Embodiment 3 filters out miRNA relevant to vincristine drug resistance using biology information technology
1, cell total rna extracts
Cell is collected, cell culture fluid is discarded, 1mL Trizol separation agent is added in each culture hole, blows and beats cell number
It is secondary to make cell cracking.It is incubated for 5min at room temperature.0.2mL chloroform reagent is added, covers pipe lid, vibrates 15s, 4 DEG C are incubated at room temperature
15min, 15000rpm are centrifuged 10min, and solution is separated into three-phase.The upper solution of colourless liquid phase is transferred to a new centrifuge tube
In.0.5mL isopropanol is added, covers pipe lid, turning upside down mixes well liquid for several times, it is incubated for 10min at room temperature, 4 DEG C
12000rpm is centrifuged l5min, abandons supernatant, and 75% ethyl alcohol is added, and oscillation washing, 4 DEG C, 7500rpm is centrifuged 5min, abandons supernatant, does
Dry 5~l0min removes ethyl alcohol.The 20 processed ddH of μ L DEPC are added2O, pipettor are blown and beaten for several times.RNA solution is taken to be divided
A260, A260/280 are measured on photometer respectively, calculates the content and purity of RNA.1 × the TAE handled with 0.2 μ L DEPC,
1% Ago-Gel, 100V constant pressure electrophoresis 30min, observation.
2, cDNA library building and sequencing
After extracting cell total rna, fragment length is recycled in the RNA of 18-30nt range, using T4RNA ligase, in segment
5 ' end add 5 ' sequence measuring joints, 3 ' end add 3 ' sequence measuring joints, then using this with connector segment as template, using random
Primer synthesizes cDNA, and 5 ' end connector primers are added and 3 ' adapter-primers carry out PCR amplification, establish the complete sequencing library of PCR.With
The 2500 high-flux sequence platform of HiSeq of high-throughput, highly sensitive Illumina company exploitation carries out HCT-8 and HCT-8/V
MiRNA sequencing.Sequencing is sequenced by Shanghai Jing Neng Biotechnology Co., Ltd.
3, statistical method
MiRNA expression quantity, which is calculated, calculates Measure Indexes (transcript per million) using TPM, and TPM formula=
(the read number that every miRNA is compared)/(the read number that sample always compares) × 106, TPM is meant that with every megabit
MiRNA expression figureofmerit is done with pairs of sequence, wherein always than expressing numerical quantity for normalizing with pairs of read number.It utilizes
DESeq software calculates separately sample expression using TPM and measures log2, meet to the miRNA of sample comparison group screening differential expression
P >=0.05 and more than or equal to 2 times differential expression ranges screen the difference miRNA between two groups.
4, the results show that compared with HCT-8 cell, the miRNA of 48 differential expressions is filtered out altogether, wherein has-miR-
663a raises 7.32 times in HCT-8/VCR cell, and difference has statistical significance (P < 0.01).
Embodiment 4 detects the expression quantity of miR-663a in HCT-8 and HCT-8/VCR cell
1, design of primers
According in miRBaSe database obtain hsa-miR-663a mature sequence (http://www.mirbase.org,
No:MI0003672): the following primer of 5 '-AGGCGGGGCGCCGCGGGACCGC-3 ' design:
Reverse transcriptase primer:
5′-CTCAACTGGTGTCGTGGAGTCGGCAATTCAGTTGCGGTCCC-3′(SEQ ID NO.2)
Quantification PCR primer:
Forward primer: 5 '-ACACTCCAGCTGGGAGGCGGGGCGCCGCGG-3 ' (SEQ ID NO.3)
Reverse primer: 5 '-TGGTGTCGTGGAGTCG-3 ' (SEQ ID NO.4)
According to U6 snRNA house-keeping gene in ncbi database (https: //www.ncbi.nlm.nih.gov/)
(Genbank:NR_004394.1) following primer is designed:
The reverse transcriptase primer of U6 snRNA: 5 '-CGCTTCACGAATTTGCGTGTCAT-3 '；(SEQ ID NO.5)
U6 forward primer: 5 '-CTCGCTTCGGCAGCACA-3 '；(SEQ ID NO.6)
U6 reverse primer: 5 '-AACGCTTCACGAATTTGCGT-3 '；(SEQ ID NO.7)
2, cell total rna extracts
Whole process wears disposable glove.With the non-disposable glassware or plastic ware of no RNA enzyme.Glassware can
To toast 4h in 150 DEG C of baking oven.Plastic ware can impregnate 10min in 0.5MNaOH, after being thoroughly rinsed completely with water
High pressure sterilization is spare.
Whole process operates on ice, prevents RNA from degrading.Take 5-10 × 106HCT-8 and HCT-8/VCR cell, from
The heart, abandons supernatant, and the Trizol reagent being pre-chilled in advance with pipette plus 1ml is blown and beaten repeatedly come lytic cell to uniform well-illuminated liquid
Afterwards, homogenised sample is placed into 5min on ice, guarantees that cell adequately cracks.200 μ l chloroforms are added, are acutely vortexed 30 seconds, room
Temperature stands 5min.4 DEG C, 12000 × g is centrifuged 15min.Careful transfer supernatant is added into a RNase-free1.5ml centrifuge tube
Isometric isopropanol mixes.4 DEG C, 12000 × g is centrifuged 15min, abandons supernatant.It is added 750 μ l, 75% ethyl alcohol, 4 degree, 12000 × g
It is centrifuged 5min, abandons supernatant.After ethyl alcohol air-dries, 45 μ l DEPC is added to handle water, is stored at room temperature 2min dissolution RNA.Denaturing electrophoretic detection
And concentration mensuration, that is, it can be used or -80 degree save.
3, DNase I processing configures following system:
| RNA | 1μg |
| 10×Reaction Buffer with MgCl2 | 1μl |
| DNase I, RNase-free | 1μl |
| DEPC handles water | To 10 μ l |
| Total volume | 10μl |
37 DEG C of water-bath 30min.65 DEG C of water-bath 10min inactivate DNase I.
4, reverse transcription
Reverse transcription reaction system (20 μ L): 5 × primer buffer, 4 μ L, reverse transcriptase 1 μ L, reverse transcriptase primer (10 μ
M) 0.5 μ L, U6 snRNA reverse transcriptase primer (10 μM), 0.5 1 μ g of μ L, RNA, adds no RNA enzyme water to mend to 20 μ L systems.It mixes, from
The heart.Reverse transcription reaction condition: 42 DEG C of incubation 15min, 85 DEG C of inactivation reverse transcriptase 5s, 4 DEG C save.
5、Real-time PCR
The reaction system 20 (μ L) of fluorescence quantitative RT-RCR: 2 × SYBR Green PCR Master Mix, 10 μ L, template
DNA1 μ L, upstream and downstream primer (10 μM) each 0.5 μ L, when distilled water is settled to the detection of 20 μ L. real time fluorescent quantitatives, forward primer point
Not with general reverse primer PCR amplification.Template cDNA is respectively that reverse transcriptase primer reverse transcription generates.
Reaction condition are as follows: 95 DEG C of denaturation 5min, 40 circulations (95 DEG C of 15s, 60 DEG C of 15s, 72 DEG C of 20s), 95 DEG C of 15s, 60
℃30s,95℃15s.It takes fluorescent value to draw solubility curve, determines the specificity of amplified production.
6, has-miR-663a expression quantity statisticallys analyze
Using U6 snRNA house-keeping gene as internal reference, using relative quantification method, the expression quantity of gene is calculated.
The expression quantity F=2 of gene—△△ct
F value is bigger, and expression quantity is higher.Wherein, (average value-of the ct of the target gene of sample to be tested is to be measured by △ △ ct=
The average value of the ct of the house-keeping gene of sample)-(average value-check sample house keeper of the ct of the target gene of control sample
The average value of the ct of gene).
The results show that has-miR-663a is raised in HCT-8/VCR cell compared with HCT-8 cell, analysis result is aobvious
Show in has-miR-663a expression quantity that in HCT-8/VCR cell, than 6.735 times of HCT-8 cell upregulation, difference has statistics
Meaning (P < 0.01) (Fig. 2).
Drug resistance after 5 cell Proliferations of embodiment-toxicity test detection HCT-8 cell transfecting has-miR-663a minics
Characteristic
When HCT-8 cell is in active growth state, 12 orifice plates are inoculated into the day before transfection, are inoculated in 12 orifice plates
HCT-8 cell suspension.It second day, transfects miRNA analogies (25nM), after transfecting 8h, vitellophag simultaneously divides disk into 96 orifice plates,
It is added vincristine (final concentration 10ng/ml, 20ng/ml, 100ng/ml, 200ng/ml, 1000ng/ml, 2000ng/ml).48h
Afterwards, 96 orifice plates are taken out, 10 μ L CCK8 solution are added to every hole and (are careful not to generate bubble in hole, they will affect OD value
Reading).Culture plate is incubated for 3h in incubator.The absorbance at 450nm is measured with microplate reader.If not measuring OD temporarily
Value, can be added 10 μ L 1%w/v SDS solution, and cover culture plate and be kept in dark place at room temperature into every hole.In for 24 hours
Measurement, absorbance will not change.
The results show that the drug resistance ability of the cell vincristine of the HCT-8 of transfection has-miR-663a analogies group is significant
Higher than untransfected group (Fig. 3), further illustrate that has-miR-663a can improve the drug resistance ability of vincristine of colon cancer cell.
After 6 cell Proliferations of embodiment-toxicity test detection HCT-8/VCR cell transfecting has-miR-663a inhibitor
Resistant characterization
Chemically synthesized has-miR-663a mortifier (being purchased from Ji Man biotechnology (Shanghai) Co., Ltd.) is transfected into
In HCT-8/VCR cell, has-miR-663a is made to express reduction in HCT-8/VCR cell.The results show that relative to control group
The HCT-8/VCR cell that can reduce of non-transfected cells, transfection has-miR-663a mortifier is shown in the drug resistance ability of vincristine
Fig. 4.
7 miRNA kit of embodiment
MiRNA kit includes miR-663a primer (such as SEQ ID NO.2, SEQ ID NO.3 and SEQ ID NO.4 institute
Show, pcr reagent (including thermal starting Taq archaeal dna polymerase (2.5U/ μ L) and its reaction buffer, dNTP
(10mM)), U6 snRNA internal control primer (SEQ ID NO.5-7) and fluorescent dye SYBR-Green 1.Utilize examination of the invention
Agent box extracts reagent and general miRNA reverse transcription reagents in conjunction with common RNA, can specifically detect miR- in tissue
The expression quantity of 663a is efficiently used for the clinical drug resistant auxiliary diagnosis of colon cancer vincristine, precisely to be treated, to be improved
Curative effect reduces side effect.
The effect detection of miRNA kit: choosing 50 colon cancer cases of Henan Prov. Tumour Hospital 2013-2015, right
Its paraffin specimen has carried out miRNA extraction, is analyzed using miRNA kit of the present invention sample, of the invention as the result is shown
MiRNA kit can specifically, effectively amplify miR-663a sequence, for statistical analysis to its expression quantity, as the result is shown
There is miR-663a expression quantity in 27 case sample miR-663a expression quantity 18.231 ± 4.121,16 samples of average out to average
It is 3.213 ± 2.134, expression quantity differs 5.67 times (P < 0.01).In conjunction with clinical therapeutic efficacy analyze, miR-663a expression quantity compared with
High sample is poor using the effect of the treatment colon cancer of vincristine, and the lower sample of expression quantity controlling using vincristine
The effect for treating colon cancer is preferable.The expression for further demonstrating miR-663a is related with the drug resistance of colon cancer vincristine.Its
Kit can be used for the drug resistance of clinical detection colon cancer vincristine, is further used for instructing clinical application and precisely control
It treats, improves curative effect, reduces side effect.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.
SEQUENCE LISTING
<110>Henan Prov. Tumour Hospital, Zhengzhou Railway Vocational and Technical College
<120>microRNA molecule miR-663a relevant to vincristine drug resistance and its application
<130> KHP171110286.7
<160> 7
<170> PatentIn version 3.5
<210> 1
<211> 22
<212> RNA
<213> miR-663a
<400> 1
aggcggggcg ccgcgggacc gc 22
<210> 2
<211> 41
<212> DNA
<213>artificial sequence
<400> 2
ctcaactggt gtcgtggagt cggcaattca gttgcggtcc c 41
<210> 3
<211> 31
<212> DNA
<213>artificial sequence
<400> 3
acactccagc tgggaggcgg ggcgccgcgg 30
<210> 4
<211> 16
<212> DNA
<213>artificial sequence
<400> 4
tggtgtcgtg gagtcg 16
<210> 5
<211> 23
<212> DNA
<213>artificial sequence
<400> 5
cgcttcacga atttgcgtgt cat 23
<210> 6
<211> 17
<212> DNA
<213>artificial sequence
<400> 6
ctcgcttcgg cagcaca 17
<210> 7
<211> 20
<212> DNA
<213>artificial sequence
<400> 7
aacgcttcac gaatttgcgt 20
Claims (5)
1. nucleotide sequence microRNA molecule miR-663a as shown in SEQ ID NO.1 or the biomaterial containing it are being made
Application in standby colon cancer vincristine drug resistance auxiliary diagnostic box；The biomaterial is carrier, host cell, transgenosis
Cell line, engineering bacteria.
2. application as described in claim 1, which is characterized in that the microRNA molecule is high in vincristine mdr cell
Expression.
3. application as described in claim 1, which is characterized in that the microRNA molecule is low in vincristine sensitive cells
Expression.
4. the inhibitor of nucleotide sequence microRNA molecule miR-663a as shown in SEQ ID NO.1 is preparing antagonism colon
Application in the drug resistant drug of cancer vincristine.
5. the specificity for PCR detection nucleotide sequence microRNA molecule miR-663a as shown in SEQ ID NO.1 is drawn
Object is to the application in preparation colon cancer vincristine drug resistance auxiliary diagnostic box, the nucleotides sequence of the specific primer pair
Column are as shown in SEQ ID NO.3-4.
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Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2016107933A3 (en) * | 2015-01-02 | 2016-08-25 | Kings College Hospital, Nhs Foundation Trust | Materials and methods for the treatment of cancers |
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2017
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| WO2016107933A3 (en) * | 2015-01-02 | 2016-08-25 | Kings College Hospital, Nhs Foundation Trust | Materials and methods for the treatment of cancers |
Non-Patent Citations (5)
| Title |
|---|
| 《Downregulation of adenomatous polyposis coli by microRNA-663 promotes odontogenic differentiation through activation of Wnt/beta-catenin signaling》;Kim Jae-Sung等;《Biochemical and biophysical research communications》;20140418;第446卷(第4期);第894-900页 |
| 《Genome-Wide Uncovering of STAT3-Mediated miRNA Expression Profiles in Colorectal Cancer Cell Lines》;Zhang Jufeng等;《Biomed research international》;20140713;第1-12页 |
| 《miR-663a inhibits colon cancer metastasis by TTC22V1》;Yantao Du等;《第八届中国肿瘤学术大会暨第十三届海峡两岸肿瘤学术会议论文汇编》;20140911;第1页 |
| 《miR-663a inhibits hepatocellular carcinoma cell proliferation and invasion by targeting HMGA2》;Huang Weizhen等;《Biomedicine &Pharmacotherapy》;20160731;第81卷;第431-438页 |
| 《miR-663a regulates growth of colon cancer cells, after administration of antimicrobial peptides, by targeting CXCR4-p21 pathway》;Kuroda Kengo等;《BMC Cancer》;20170107;第17卷;第1-10页 |
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