CN103721268A - Application of miR-150 inhibitor in preparation of colorectal cancer drug resistance reversal agent - Google Patents
Application of miR-150 inhibitor in preparation of colorectal cancer drug resistance reversal agent Download PDFInfo
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Abstract
The invention belongs to the field of biological medicine, relates to a drug resistance reversal agent, and in particular relates to a drug resistance reversal agent for reversing the drug resistance in colorectal cancer treatment. The invention provides an application of a miR-150 inhibitor in the preparation of a colorectal cancer drug resistance reversal agent, especially an application of the miR-150 inhibitor in the preparation of an HCT-116 series and/or HT-29 series colorectal cancer cell drug resistance reversal agent.
Description
Technical field
The invention belongs to biomedicine field, relate to a kind of reversal agent of drug resistance, more particularly, relate to a kind of reversal agent of drug resistance that reverses treatment of colorectal cancer drug resistance.
Background technology
Colorectal cancer (Colorectal Cancer, CRC) is the common malignant tumor in third place in the world.In China, along with improving constantly of living standards of the people, the change of dietary habit and aged tendency of population, colorectal cancer incidence rate rapid growth.Existing treatment of colorectal cancer scheme, except operative treatment, also needs chemoradiotherapy as auxiliary treatment means.But due to the existence of tumor to the drug resistance of medicine, have a strong impact on the effect for the treatment of of colorectal cancer.
MicroRNA is the endogenous non-coding microRNA that a class is about 18～25 nucleotide, participates in and regulates and controls nearly all cytobiology process, comprises sequential growth, apoptosis, lipid metabolism, neuronal development, cell differentiation, cell proliferation etc. are in interior various physiological processes.It suppresses the expression of target gene by being combined with the non-coding region of target gene.It is predicted, in human genome, may exist 1/3rd protein coding gene to be nearly subject to the regulation and control of microRNA.In the tumor cell of mankind's majority, all find the unconventionality expression of microRNA, and in different cancer generating processes, be nearly all accompanied by the expression change of microRNA.More than 50% mankind microRNA gene is positioned at tumor-related gene region, as fragile site, and loss of heterozygosity district, amplification region or common fracture zone.Appear in these and the closely-related mutable gene group of tumor microRNA gene high frequency, pointing out it with tumor, to occur with development closely related.In colorectal cancer, the unconventionality expression of microRNA and the generation of tumor and development significant correlation, some specific microRNA, as the important biomolecule molecule that regulates chemosensitivity, has participated in colorectal cancer Chemoresistance simultaneously.Current research shows microRNA-150(miR-150) unconventionality expression in numerous tumors the heavy role who plays the part of in a series of biochemical processes.Researcher is found thousands of potential target genes by bioinformatics method, and confirms that by further experiment c-Myb, MUC4, ZEB1, AKT2, DKC1, EGR2, P2X7 and p53 are the direct target genes of miR-150.By analyzing their regulatory pathway and in the effect of cell, we infer them, and wherein the drug resistance of some target gene and chemotherapeutics is closely related.As p53 albumen plays key effect in the propagation of the apoptosis-induced approach of p53 and inhibition cell.EGR2 in PTEN growth inhibited path as tumor-inhibiting factor, thereby it by regulation and control Fas ligand the expression cell death inducing at T cell.EGR2 also can affect in the activity of tumor cell the activity that relies on the protein induced apoptosis pathway of P53 in addition.P2X7 acts on tumor-inhibiting factor and participates in the apoptosis-induced approach of caspase-9 mitochondrion.Oncogene c-myb is proved the propagation that can strengthen cell and the apoptosis that suppresses cell in cell.
At present, miR-150 is still comparatively short of in the research of colorectal cancer.Inventor's research in earlier stage finds that miR-150 is closely related at unconventionality expression and the tumor drug resistance of colorectal cancer clinical patient tissue.Yet, because the pathogenesis of cancer is complicated, the increment of various cancers hypotype and apoptosis mechanism and regulatory pathway thereof are also different, therefore, although it is relevant to numerous tumor regulatory pathway that the unconventionality expression of miR-150 has been known, but, complexity due to the apoptosis pathway of the even identical tumor different subtype of different tumors, if the common index of the cancer patient's regulatory pathway using miR-150 as multiple different subtype, its specificity and accuracy rate are all lower, therefore, at present still very limited in Drug Resistance Detection and the application aspect drug resistance inversion of cancer for miR-150.
Summary of the invention
The object of the present invention is to provide the detectable for particular type colorectal cancer patients drug resistance that a kind of accuracy rate is high.
Another object of the present invention is to provide a kind of inversion agent of effective reverse colorectal cancer drug resistance.
A further object of the present invention is to provide a kind of inversion agent for particular type colorectal cancer drug resistance.
Invention is achieved through the following technical solutions above-mentioned purpose:
First, aspect the detectable of colorectal cancer patients drug resistance, provided by the present invention is a kind of drug resistance detectable---miR-150 detectable for HCT-116 and/or HT-29 tying intestinal cancer.
Preferably, the invention provides a kind of accuracy rate, sensitivity far away higher than the drug resistance detectable for HT-29 type colon cancer---the miR-150 detectable of prior art.
Although research before this shows, miR-150 drug resistance patient's patient in colorectal cancer patients expression is higher than sensitive patients.Yet this result is not made differentiation for dissimilar cancer patient.After the inventor distinguishes dissimilar cancer cell, find that for example, mdr cell miR-150 expression contrasts non-mdr cell in some type (SW620), not the raising of highly significant.And in HCT-116 and HT-29, especially, in HT-29, mdr cell miR-150 expression contrasts the raising that non-mdr cell demonstrates highly significant.
Therefore, using miR-150 detectable as HCT-116 and/or the drug resistance detectable of HT-29 tying intestinal cancer, especially, as the drug resistance detectable of HT-29 type colon cancer, accuracy rate and sensitivity will improve greatly.
Except this, invention provides a kind of inversion agent---miR-150 inhibitor of effective reverse colorectal cancer drug resistance.Described colorectal cancer is the colorectal cancer of high expressed miR-150.
Preferably, invention provides the application of miR-150 inhibitor on preparation HCT-116 system and/or HT-29 tying rectum cancer cell reversal agent of drug resistance.Particularly preferably, be HT-29 tying rectum cancer cell.
Inventor infers that according to the result of study in early stage miR-150 may pass through the expression of negative regulation c-myb, EGR2, P2X7 and p53, thereby affects generation and the development of chemotherapeutics drug resistance.
Further, inventor's discovery, when using cell cycle medicine to carry out chemotherapy, miR-150 plays anti-apoptotic effect in HCT-116 and/or HT-29 cell.Its possible mechanism is due to miR-150 negative regulation oncogene c-myb, suppress cell proliferation, and cell enlargement speed can allow cell still less enter the DNA replication dna phase that medicine works slowly, has increased the drug resistance to medicine, thereby escape from chemotherapy.Colorectal cancer cell is that HCT-116 and HT-29 cross the propagation that expression miR-150 can suppress cell, causes the increase of cells resistance to make the Reduced susceptibility of cell to medicine.When existing in the situation of miR-150 inhibitor, can eliminate the anti-apoptotic effect of miR-150, this treatment to drug resistance patient is particularly important.
Described miR-150 inhibitor can be common arbitrary inhibitor, for example, can and miR-150 and its target between combination there is interactional sequence; Include the nucleotide sequence that can be combined with miR-150; Comprise with at least 6 nucleotide of the sequence of ripe miR-150 and substantially go up complementary nucleotide sequence; Comprise and the Seed Sequences of miR-150 complementary nucleotide sequence substantially; Comprise the nucleotide sequence that can be combined with ripe miR-150 under physiological condition; Can reduce value machine of ripe miR-150 level in cell etc.
Preferably, described drug resistance is by described medicine, to disturb P53 to rely on apoptosis-induced approach to be caused.Preferably, described medicine refers to 5-fluorouracil (5-FU) class medicine.
5-fluorouracil as colorectal cancer patients main chemotherapy medication continued to use nearly half a century, its main toxic action can make an explanation from following this two aspect:
(1) 5-FU can activate in vivo and generate after 5-fluorouracil Deoxydization nucleotide, is combined with the sweet acid enzyme of breast, suppresses the activity of this enzyme, makes the sweet azymia of deoxidation breast, DNA replication dna obstacle.(2) 5-FU can metabolism be also 5-fluorouracil nucleoside, as pseudo-metabolite form, is incorporated in RNA, affects RNA function and protein synthesis.But due to the existence of tumor to Drug tolerance, chemotherapy resistance has become the major obstacle of effective treatment Patients with Colorectal Cancer.
Inventor's discovery, in colorectal cancer mdr cell, the existence of miR-150 makes the use of 5-FU can not improve the apoptosis rate of cell.And the use of 5-FU also can obviously promote the expression of miR-150, the degree of lifting is relevant to the concentration of 5-FU, and this has further worsened the drug resistance of cancer cell.Especially in HCT-116 and HT-29 cell, this deterioration degree is more outstanding.Therefore, when adopting 5-FU class medicine to carry out chemotherapy, the existence of miR-150 inhibitor is even more important, and it can suppress crossing of miR-150 expresses, and destroys the anti-apoptotic effect of miR-150, and reversing drug resistance patient's drug resistance is played to particularly important effect.
Compared with prior art, the detectable of the colorectal cancer patients drug resistance for particular type of the present invention, has incomparable before this accuracy rate and sensitivity by the drug resistance diagnosis to this particular type.Thereby for the Rational Decision of follow-up therapeutic scheme provides solid foundation.
The present invention for the inversion agent of colorectal cancer drug resistance of particular type, can effectively reverse more targetedly patient's drug resistance, especially in 5-FU chemotherapy, this inversion agent has fundamentally been contained the resistance mechanism of the caused cell of 5-FU class medicine, has reversed the drug resistance of cell.In HCT-116 and HT-29, adopt miR-150 inhibitor to reach respectively 30.8% and 37.7% as the effective percentage of resistance to 5-FU inversion agent.
Accompanying drawing explanation
Fig. 1 shown miR-150 as drug resistance index the differential expression in different colorectal cancer cells, and the impact of miR-150 on HCT-116 and HT-29 cells resistance;
Wherein, (a) miR-150 is at Caco-2, HCT-116, the expression in HT-29 and SW620 cell (
*p<0.05); (b) transfection the expression of the HCT-116 of RNA analogies miR-150 and the miR-150 of HT-29 cell; (c, d) under a series of 5-FU concentration (0 μ M, 2 μ M, 4 μ M, 6 μ M, 8 μ M, 10 μ M, 12 μ M) cross and express the HCT-116 of miR-150 and the cytotoxic effect that SW620 reduces 5-FU induction.
Fig. 2 is that miR-150 crosses the impact of expressing HCT-116 and HT-29 cell proliferation and apoptosis;
(a, b) crosses and expresses the propagation that miR-150 suppresses HCT-116 and HT-29 cell; (c) cross and express the apoptosis that miR-150 can reverse 5-FU induction in HCT-116 and HT-29 cell; (d) cross and express the quantized result that miR-150 can reverse the apoptosis of 5-FU induction in HCT-116 and HT-29.
Fig. 3 has shown that 5-FU has improved the expression of miR-150 in HCT-116 and HT-29 cell;
The expression of (a, b) HCT-116 and HT-29 (0 μ M, 2 μ M, 4 μ M, 6 μ M, 8 μ M, 10 μ M) miR-150 under the processing of variable concentrations 5-FU; (c) 5-FU has improved the expression of miR-150 at HCT-116 and HT-29 under the concentration of 10 μ M, but on not impact of the expression of the miR-150 of SW480 cell (
*p<0.05).
The specific embodiment
The following specific embodiment is the further explaination to technical solution of the present invention, and unrestricted.
The relation of miR-150 expression and drug resistance in the dissimilar colorectal cancer cell of embodiment 1 system
The cultivation of cell:
Colorectal cancer cell is Caco-2, HCT-116, HT-29 and SW620 are preserved by Zhongshan University's gastroenterology's institute, Caco-2 is with containing 10% hyclone (Invitrogen, Grand Island, NY, USA) DMEM(Gibco, China) culture medium, HCT-116, HT-29 and SW620 are with containing 10% hyclone (Invitrogen, Grand Island, NY, USA) RPMI1640(Gibco, China) culture medium, is placed in containing 5% carbon dioxide, the incubator of 37 ℃ of constant temperature humidities and cultivates.
5-FU exposure test:
By 5-FU(Sigma, China) with DMSO, to be mixed with concentration be that the storage liquid of 0.1M is standby to powder.Cell is with 0.25 * 10
6the density in individual/hole is cultivated 24 hours after being inoculated in 6 orifice plates, adds 5-FU to make final concentration be respectively 2 μ M, 4 μ M, and 6 μ M, 8 μ M, 10 μ M, take solvent DMSO as blank, cultivate respectively 24,48,72 hours.
The detection of miR-150 expression:
According to the step in manufacturer's description of TRIZOL reagent (Invitrogen, USA), extract cell total rna.Step is as follows: (1) is inhaled and abandoned culture medium, with the PBS of 1ml, rinses twice, and every hole adds TRIZOL reagent 1ml, and room temperature is placed 5min; (2) cell pyrolysis liquid is all transferred in the EP pipe of 1.5ml, added 0.2ml chloroform, concuss 15s, standing 5min under room temperature, 12,000g, 4 ℃ of centrifugal 15min; (3) get upper water phase transfer in the new centrifuge tube without RNase of another one, add isopyknic isopropyl alcohol, softly mix, room temperature is placed 10min, 12,000g, 4 ℃ of centrifugal 15min; (4) abandon supernatant, add 75% washing with alcohol RNA precipitation, 7,500g, 4 ℃ of centrifugal 5min, abandon supernatant, and air drying RNA precipitates 10min, with DEPC water dissolution RNA.With All-in-One miRNA qRT-PCR Detection Kit (Gene Copoeia, China) test kit, carry out RT-PCR amplification.Gained quantitative data is derived by 7500software v2.0.6 (Applied Biosystems), the threshold cycle (C of each sample
t) value represents the expression of institute's cls gene, by Δ Δ C
tthe value that method is calculated represents the difference of miR-150 and reference gene, finally by 2
-Δ Δ Ctmethod deal with data, show that data represent the expression of miR-150.
Result as shown in Figure 1.At HCT-116, in HT-29 colorectal cancer cell system, cross expression miR-150 and tumor cell drug resistance closely related.Inventor's microRNAs gene chip result shows that miR-150 drug resistance patient's patient in colorectal cancer patients expression is 2.28 times of (Dou of sensitive patients, R., et al., Multi-microarray identifies lower AQP9expression in adjuvant chemotherapy nonresponders with stage III colorectal cancer.Cancer Lett, 2013.336 (1): p.106-13.).Early-stage Study shows that different colorectal cancer cells are different to the chemosensitivity degree of 5-FU, and the drug resistance relation that is obtained them by IC50 (half-inhibition concentration) is Caco-2, HCT-116, HT-29 and SW620 from small to large.In order further to inquire into the relation of miR-150 and chemotherapy resistance, with reference to inventor's early-stage Study, from each colorectal cancer cell, be measured IC50(half-inhibition concentration), in these four kinds of cell lines, detect the expression of miR-150, in this experiment, the Δ CT value of Caco-2 is reference value, and U6 is internal reference.Data show (Fig. 1),
Result has demonstrated the result outside aforementioned institute expection.Except SW620, HCT-116 and HT-29 are along with the increase of cells resistance, and the expression of miR-150 increases thereupon, and especially for HT-29, the expression of miR-150 and the drug resistance degree of association are far away higher than other cells (Fig. 1).
And then inventor uses analogies miR-150 and miR-NC transient transfection HCT-116 and the HT-29 of microRNA, and microRNA analogies ultimate density is 50nM.After transfection 4-6 hour, change not containing antibiotic fresh complete medium.In order to detect the transfection efficiency of miR-150, transfectional cell is cultivated and within 3 days, is extracted afterwards the expression that cell total rna carries out RT-PCR detection miR-150, and the expression of the miR-150 of the cell (processed group) of result demonstration transfection miR-150 is apparently higher than matched group (Fig. 1 b).After determining transfection efficiency, we carry out the Drug toxicity trails of 5-FU, by transient transfection HCT-116 and the HT-29 of microRNA analogies, cultivating and changing concentration in 24 hours is the 5-FU of 8 μ M, cultivates after 3 days with WST-1 detection cell absorbance.Result of the test confirmed to cross express the HCT-116 of miR-150 and HT-29 compare transfection the cell (miR-NC) of contrast micoRNA analogies more can resist the inducing action of 5-FU.(Fig. 1 c, d)
Visible, at colorectal cancer cell, be in HCT-116 and HT-29, to cross expression miR-150 can cause the more resistance to 5-FU(Fig. 1 of cell).But this situation is not all applicable to all colorectal cancer cells.
Aspect the judgement of drug resistance, different colorectal cancer cell systems is distinguished, by miR-150 detectable, as HCT-116 or HT-29 colorectal cancer cell, be specifically the detectable of drug resistance, the accuracy rate that can be greatly improved, reaches respectively 78.5% and 85.9%.
The anti-apoptotic effect of embodiment 2miR-150 in dissimilar colorectal cancer cell is
MiR-150 transient transfection HCT-116 and HT-29:
After cell is inoculated in culture plate 24 hour cell density and reaches 50%, according to manufacturer, illustrate with liposome Lipofectamine2000 (Invitrogen, USA) be analogies miR-150 and miR-NC transient transfection HCT-116 and the HT-29 of carrier with microRNA, microRNA analogies ultimate density is 50nM.After transfection 4-6 hour, change not containing antibiotic fresh complete medium.In order to verify transfection effect, arrange in addition and within three days, extract total RNA after experimental cell transfection and verify that by real-time quantitative PCR miR-150 crosses expression effect.
Cell proliferation test:
Use WST-1 cell proliferation and cytotoxicity detection kit (Roche Applied Science, USA) to carry out cell proliferation experiment.After being inoculated in the cell transfecting of 96 orifice plates, cultivate respectively 24,48,72 hours, culture medium is changed once for every two days, when reaching, cultured cell sets after natural law, fast the WST-1 of 10 μ l is added in hole, cultivates after 30min, use microplate reader (BioRad, China) detect absorbance, until 90min, microplate reader detects and sets wavelength is 450nm, and reference wavelength is 630nm.
Apoptosis test:
After cell transfecting 24 hours, changing final concentration is the complete medium (not containing antibiotic) of the 5-FU of 8 μ M, take DMSO as solvent control group, continue to cultivate after 48 hours with trypsin digestion cell, with PBS(Phosphate Buffered Saline) re-suspended cell 3 times to be to remove culture medium completely, use (Annexin V-FITC apoptosis Detection kit, BD Biosciences, USA) test kit to carry out cell apoptosis assay.(1) according to 1 * 10
6the concentration of/ml is resuspended with 1 * Binding Buffer by the cell after rinsing; (2) cell suspension of getting 100 μ l, in flow cytometer detection pipe, then adds the APC Annexin V of 5 μ l and the PI mix homogeneously of 5 μ l, and greenhouse lucifuge is hatched 20min; (3) according to the cell volume of every 50 μ l, add Intrapred reagent I mix homogeneously, room temperature lucifuge is hatched 15min; (4) add 1 * Binding Buffer of 400 μ l resuspended, in 1 hour, carry out apoptosis detection.Data are calculated with flow cytometry software BD FACSDiva Software v6.1.3.
Statistical analysis:
Data are drawn and are analyzed and use respectively Graphpad Prism (version5.0) and SPSS (version18.0) software.Minimum repetition twice, the two sample independence T test evaluation data statistics meaning of all tests, setting P<0.05 difference has statistical significance.
In order to study the effect of miR-150 in colorectal cancer, we have carried out cell proliferation experiment.We find that miR-150 can suppress the propagation of HCT-116 and HT-29 cell, and result is as shown in Figure 2 at the 5th day, and the suppression ratio of HCT-116 experimental group is 63.7% ± 8.5%, and matched group is that 27.4 ± 14% (Fig. 2 a).The suppression ratio of HT-29 experimental group is 65.1% ± 10.9%, and matched group is 29.2 ± 9.1% (Fig. 2 b).
Apoptosis is escaped and is played an important role to the formation of chemotherapeutics tolerance and in developing in tumor, and for this reason, we carry out cell apoptosis assay.We use miR-150 and miR-NC transfection HCT-116 and HT-29, continue to cultivate after 48 hours, with detecting apoptosis kit detection cell apoptosis rate, discovery is compared with matched group, the apoptosis rate of the cell of transfection miR-150 all declines to some extent, and fall reaches respectively 22.3%(HCT-116) and 9.6%(HT-29), miR-150 has played anti-apoptosis (Fig. 2 c) here, Fig. 2 e, what 2f showed is exactly the quantitative analysis of apoptosis rate.In order further to confirm the effect of the anti-apoptosis of miR-150, after transfection, we detect its apoptosis rate for 48 hours again by 5-FU and the co-culture of cells of 8 μ M, and solvent in contrast.The apoptosis rate that discovery has added the cell of 5-FU has no and increases or than lower (Fig. 2 d of the apoptosis rate of matched group, 2f), therefore we infer this phenomenon may with miR-150 negative regulation apoptosis inducing factor p53(Zhang, N., X.Wei, and L.Xu, miR-150promotes the proliferation of lung cancer cells by targeting P53.FEBS Lett, 2013.587 (15): p.2346-51.), EGR2(Wu, Q., et al., MiR-150promotes gastric cancer proliferation by negatively regulating the pro-apoptotic gene EGR2.Biochem Biophys Res Commun, p.340-5.) and P2X7 (Zhou 2010.392 (3):, L., et al., MicroRNAs miR-186and miR-150down-regulate expression of the pro-apoptotic purinergic P2X7receptor by activation of instability sites at the3 '-untranslated region of the gene that decrease steady-state levels of the transcript.J Biol Chem, 2008.283 (42): p.28274-86.) apoptosis inhibit is relevant.
The result of comprehensive above-mentioned two embodiment, adopts miR-150 inhibitor as the reversal agent of drug resistance of HCT-116 and HT-29, eliminates the apoptosis inhibitory action of miR-150, thereby reverses cells resistance (resistance to 5-FU).Inventor adopts the reverse HCT-116 of miR-150 inhibitor and the effective percentage of HT-29 drug resistance aspect to reach respectively 30.8% and 37.7%.
The rise effect of embodiment 3 5-FU to miR-150 expression in HCT-116 and HT-29
Studies have reported that 5-FU can regulate and control the expression of some microRNA, as in colorectal cancer, thereby cause that the medicine of DNA damage can raise miR-34a, miR-16-1 by raising the expression of P53, miR-143, the expression of miR-145 and miR-206.5-FU also can lower MiR-200b, the expression of miR-210 and miR-224 simultaneously.In order to verify the 5-FU regulating and controlling effect possible to miR-150, we process the expression of HCT-116 and HT-29 detection miR-150 with the 5-FU of a series of concentration.
Experimental result is as shown in Figure 3 in HCT-116 and HT-29, and 5-FU can obviously promote the expression (Fig. 3 a, 3b) of miR-150.The expression that further experimental results show that 5-FU rise miR-150 has cell-specific, and it can significantly improve the expression of miR-150 in HCT-116 and HT-29, but does not affect miR-150 at the expression of SW480.
Result by Fig. 2 and Fig. 3, miR-150 can suppress the apoptosis of 5-FU induction, and 5-FU also can promote the expression of miR-150 simultaneously, further worsen the drug resistance of cell and hindered carrying out smoothly for the treatment of, visible, miR-150 inhibitor has particularly outstanding meaning and effect aspect the cells resistance reverse in 5-FU chemotherapy process, is particularly useful for the colorectal cancer 5-FU drug resistance inversion of HCT-116 and HT-29 system.
Claims (9)
- The application of 1.miR-150 inhibitor on preparation colorectal cancer reversal agent of drug resistance.
- 2. application as claimed in claim 1, is characterized in that described colorectal cancer is the colorectal cancer of high expressed miR-150.
- 3. application as claimed in claim 1, is characterized in that described colorectal cancer is HCT-116 system and/or HT-29 system.
- 4. application as claimed in claim 1, is characterized in that described colorectal cancer is HT-29 system.
- 5. application as claimed in claim 1, is characterized in that described drug resistance is by described medicine, to disturb P53 to rely on apoptosis-induced approach to be caused.
- 6. application as claimed in claim 1, is characterized in that described medicine is cell cycle medicine.
- 7. application as claimed in claim 1, is characterized in that described medicine 5-FU class medicine.
- The application of 8.miR-150 detectable in preparation colorectal cancer patients drug resistance detectable, described rectal cancer is HCT-116 system and/or HT-29 system.
- 9. application as claimed in claim 8, is characterized in that described rectal cancer is HT-29 system.
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| CN107569471A (en) * | 2017-08-22 | 2018-01-12 | 深圳市龙岗区耳鼻咽喉医院 | A kind of preparation method of the nano-particles of microRNA 150 |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN104267188A (en) * | 2014-08-28 | 2015-01-07 | 汪建平 | Applications of related preparations aiming at MSK1 gene in preparation of 5-FU drug resistance detection reagent and 5-FU drug resistance reversal agent |
| CN104267188B (en) * | 2014-08-28 | 2015-12-30 | 汪建平 | For the application of related preparations in preparation 5-FU Resistance detection reagent and 5-FU reversal agent of drug resistance of MSK1 gene |
| CN105734121A (en) * | 2014-12-29 | 2016-07-06 | 中国人民解放军第三军医大学第附属医院 | Application of aldehyde dehydrogenases 1A3 and encoding gene thereof as target for preventing and treating invasion and metastasis of colorectal cancer |
| CN107569471A (en) * | 2017-08-22 | 2018-01-12 | 深圳市龙岗区耳鼻咽喉医院 | A kind of preparation method of the nano-particles of microRNA 150 |
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