CN116103400A - LncRNA-ENST00000446135在制备预测AML预后试剂盒中的应用 - Google Patents
LncRNA-ENST00000446135在制备预测AML预后试剂盒中的应用 Download PDFInfo
- Publication number
- CN116103400A CN116103400A CN202211743017.7A CN202211743017A CN116103400A CN 116103400 A CN116103400 A CN 116103400A CN 202211743017 A CN202211743017 A CN 202211743017A CN 116103400 A CN116103400 A CN 116103400A
- Authority
- CN
- China
- Prior art keywords
- lncrna
- prognosis
- kit
- aml
- cells
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 208000031261 Acute myeloid leukaemia Diseases 0.000 title claims abstract description 68
- 238000004393 prognosis Methods 0.000 title claims abstract description 33
- 238000002360 preparation method Methods 0.000 title claims abstract description 8
- 230000014509 gene expression Effects 0.000 claims abstract description 25
- 210000001185 bone marrow Anatomy 0.000 claims abstract description 10
- 238000010837 poor prognosis Methods 0.000 claims abstract description 4
- 208000033776 Myeloid Acute Leukemia Diseases 0.000 claims description 57
- 239000003153 chemical reaction reagent Substances 0.000 claims description 10
- 210000005087 mononuclear cell Anatomy 0.000 claims description 8
- 238000001514 detection method Methods 0.000 claims description 7
- 238000011529 RT qPCR Methods 0.000 claims description 6
- 238000003757 reverse transcription PCR Methods 0.000 claims description 5
- 238000002123 RNA extraction Methods 0.000 claims description 4
- 230000000694 effects Effects 0.000 abstract description 6
- 210000004027 cell Anatomy 0.000 description 83
- 241000700605 Viruses Species 0.000 description 22
- 238000001890 transfection Methods 0.000 description 19
- 239000000243 solution Substances 0.000 description 14
- 239000001963 growth medium Substances 0.000 description 13
- RXWNCPJZOCPEPQ-NVWDDTSBSA-N puromycin Chemical compound C1=CC(OC)=CC=C1C[C@H](N)C(=O)N[C@H]1[C@@H](O)[C@H](N2C3=NC=NC(=C3N=C2)N(C)C)O[C@@H]1CO RXWNCPJZOCPEPQ-NVWDDTSBSA-N 0.000 description 12
- 239000002609 medium Substances 0.000 description 11
- 238000004806 packaging method and process Methods 0.000 description 10
- 241000713666 Lentivirus Species 0.000 description 9
- 239000006228 supernatant Substances 0.000 description 9
- 239000012091 fetal bovine serum Substances 0.000 description 8
- 239000010410 layer Substances 0.000 description 8
- 239000007788 liquid Substances 0.000 description 8
- 238000002156 mixing Methods 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- 230000004083 survival effect Effects 0.000 description 8
- 239000013612 plasmid Substances 0.000 description 7
- 238000011282 treatment Methods 0.000 description 7
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 6
- 238000005119 centrifugation Methods 0.000 description 6
- 238000010790 dilution Methods 0.000 description 6
- 239000012895 dilution Substances 0.000 description 6
- 208000015181 infectious disease Diseases 0.000 description 6
- 108090000623 proteins and genes Proteins 0.000 description 6
- 229950010131 puromycin Drugs 0.000 description 6
- 239000000523 sample Substances 0.000 description 6
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 5
- 230000006907 apoptotic process Effects 0.000 description 5
- 238000006243 chemical reaction Methods 0.000 description 5
- 238000002512 chemotherapy Methods 0.000 description 5
- 238000012258 culturing Methods 0.000 description 5
- 239000003550 marker Substances 0.000 description 5
- 239000013598 vector Substances 0.000 description 5
- 239000012980 RPMI-1640 medium Substances 0.000 description 4
- 108091046869 Telomeric non-coding RNA Proteins 0.000 description 4
- 239000006285 cell suspension Substances 0.000 description 4
- 239000002299 complementary DNA Substances 0.000 description 4
- 238000003745 diagnosis Methods 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 210000004698 lymphocyte Anatomy 0.000 description 4
- 230000035755 proliferation Effects 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 3
- 239000013614 RNA sample Substances 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 238000000605 extraction Methods 0.000 description 3
- 238000011134 hematopoietic stem cell transplantation Methods 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 238000001638 lipofection Methods 0.000 description 3
- 238000000034 method Methods 0.000 description 3
- 230000002018 overexpression Effects 0.000 description 3
- 239000012096 transfection reagent Substances 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- 108050008874 Annexin Proteins 0.000 description 2
- 102000000412 Annexin Human genes 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 108020004414 DNA Proteins 0.000 description 2
- 229920001917 Ficoll Polymers 0.000 description 2
- HTTJABKRGRZYRN-UHFFFAOYSA-N Heparin Chemical compound OC1C(NC(=O)C)C(O)OC(COS(O)(=O)=O)C1OC1C(OS(O)(=O)=O)C(O)C(OC2C(C(OS(O)(=O)=O)C(OC3C(C(O)C(O)C(O3)C(O)=O)OS(O)(=O)=O)C(CO)O2)NS(O)(=O)=O)C(C(O)=O)O1 HTTJABKRGRZYRN-UHFFFAOYSA-N 0.000 description 2
- 229920000209 Hexadimethrine bromide Polymers 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- 108020005198 Long Noncoding RNA Proteins 0.000 description 2
- 208000036142 Viral infection Diseases 0.000 description 2
- 230000005856 abnormality Effects 0.000 description 2
- 238000002835 absorbance Methods 0.000 description 2
- 230000001464 adherent effect Effects 0.000 description 2
- 230000003321 amplification Effects 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- 238000001516 cell proliferation assay Methods 0.000 description 2
- 230000001413 cellular effect Effects 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 238000001784 detoxification Methods 0.000 description 2
- 238000007865 diluting Methods 0.000 description 2
- 230000003828 downregulation Effects 0.000 description 2
- 230000002068 genetic effect Effects 0.000 description 2
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 2
- 229960002897 heparin Drugs 0.000 description 2
- 229920000669 heparin Polymers 0.000 description 2
- 238000011081 inoculation Methods 0.000 description 2
- 230000000670 limiting effect Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 239000013642 negative control Substances 0.000 description 2
- 238000003199 nucleic acid amplification method Methods 0.000 description 2
- 239000013600 plasmid vector Substances 0.000 description 2
- 230000000306 recurrent effect Effects 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 238000013517 stratification Methods 0.000 description 2
- 239000000725 suspension Substances 0.000 description 2
- 229910021642 ultra pure water Inorganic materials 0.000 description 2
- 239000012498 ultrapure water Substances 0.000 description 2
- 230000009385 viral infection Effects 0.000 description 2
- 102100036009 5'-AMP-activated protein kinase catalytic subunit alpha-2 Human genes 0.000 description 1
- 206010069754 Acquired gene mutation Diseases 0.000 description 1
- 108091032955 Bacterial small RNA Proteins 0.000 description 1
- 108010077544 Chromatin Proteins 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 208000034951 Genetic Translocation Diseases 0.000 description 1
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 1
- 208000002250 Hematologic Neoplasms Diseases 0.000 description 1
- 101000783681 Homo sapiens 5'-AMP-activated protein kinase catalytic subunit alpha-2 Proteins 0.000 description 1
- 102100034343 Integrase Human genes 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 102220612388 Mitogen-activated protein kinase kinase kinase 1_C15S_mutation Human genes 0.000 description 1
- 102100031455 NAD-dependent protein deacetylase sirtuin-1 Human genes 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 1
- 238000003559 RNA-seq method Methods 0.000 description 1
- 101710141795 Ribonuclease inhibitor Proteins 0.000 description 1
- 229940122208 Ribonuclease inhibitor Drugs 0.000 description 1
- 102100037968 Ribonuclease inhibitor Human genes 0.000 description 1
- 102000006382 Ribonucleases Human genes 0.000 description 1
- 108010083644 Ribonucleases Proteins 0.000 description 1
- 108010041191 Sirtuin 1 Proteins 0.000 description 1
- 108091027967 Small hairpin RNA Proteins 0.000 description 1
- 108020004459 Small interfering RNA Proteins 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 230000010100 anticoagulation Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 239000012148 binding buffer Substances 0.000 description 1
- 230000033228 biological regulation Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 238000004364 calculation method Methods 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 238000004113 cell culture Methods 0.000 description 1
- 239000012930 cell culture fluid Substances 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 230000030833 cell death Effects 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 230000007726 cellular glucose metabolism Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 229940044683 chemotherapy drug Drugs 0.000 description 1
- 210000003483 chromatin Anatomy 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000002247 constant time method Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 230000002559 cytogenic effect Effects 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 230000004069 differentiation Effects 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000001035 drying Methods 0.000 description 1
- 230000008482 dysregulation Effects 0.000 description 1
- 230000001973 epigenetic effect Effects 0.000 description 1
- 210000003237 epithelioid cell Anatomy 0.000 description 1
- 210000003743 erythrocyte Anatomy 0.000 description 1
- 230000005284 excitation Effects 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 230000002349 favourable effect Effects 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 210000003714 granulocyte Anatomy 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 230000009033 hematopoietic malignancy Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 238000001727 in vivo Methods 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 230000002452 interceptive effect Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000002372 labelling Methods 0.000 description 1
- 230000003211 malignant effect Effects 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000037353 metabolic pathway Effects 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 230000011987 methylation Effects 0.000 description 1
- 238000007069 methylation reaction Methods 0.000 description 1
- 108091086222 miR-520c stem-loop Proteins 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 210000003643 myeloid progenitor cell Anatomy 0.000 description 1
- 108091027963 non-coding RNA Proteins 0.000 description 1
- 102000042567 non-coding RNA Human genes 0.000 description 1
- 238000010606 normalization Methods 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 238000012858 packaging process Methods 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 210000002381 plasma Anatomy 0.000 description 1
- 238000007747 plating Methods 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- XJMOSONTPMZWPB-UHFFFAOYSA-M propidium iodide Chemical compound [I-].[I-].C12=CC(N)=CC=C2C2=CC=C(N)C=C2[N+](CCC[N+](C)(CC)CC)=C1C1=CC=CC=C1 XJMOSONTPMZWPB-UHFFFAOYSA-M 0.000 description 1
- 239000011535 reaction buffer Substances 0.000 description 1
- 230000002829 reductive effect Effects 0.000 description 1
- 230000022983 regulation of cell cycle Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 239000003161 ribonuclease inhibitor Substances 0.000 description 1
- 210000002966 serum Anatomy 0.000 description 1
- 230000035939 shock Effects 0.000 description 1
- 239000002356 single layer Substances 0.000 description 1
- 239000004055 small Interfering RNA Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 230000037439 somatic mutation Effects 0.000 description 1
- 230000003595 spectral effect Effects 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 238000011476 stem cell transplantation Methods 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 230000032258 transport Effects 0.000 description 1
- 238000011269 treatment regimen Methods 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 238000005199 ultracentrifugation Methods 0.000 description 1
- 238000011144 upstream manufacturing Methods 0.000 description 1
- 230000003612 virological effect Effects 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6883—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material
- C12Q1/6886—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for diseases caused by alterations of genetic material for cancer
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/118—Prognosis of disease development
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/158—Expression markers
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q2600/00—Oligonucleotides characterized by their use
- C12Q2600/178—Oligonucleotides characterized by their use miRNA, siRNA or ncRNA
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Engineering & Computer Science (AREA)
- Immunology (AREA)
- Pathology (AREA)
- Analytical Chemistry (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Wood Science & Technology (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Hospice & Palliative Care (AREA)
- Biophysics (AREA)
- Oncology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
本发明属于生物技术领域,具体涉及LncRNA‑ENST00000446135在制备预测AML预后试剂盒中的应用。本发明首次发现AML患者骨髓中LncRNA‑ENST00000446135的表达水平与AML患者的预后相关,当其表达水平CT值≥0.655时,指示AML患者预后良好;当其表达水平CT值<0.655时,指示AML患者预后不良。本发明试剂盒对AML患者的预后预测效果较好,具有广阔的临床应用前景。
Description
技术领域
本发明属于生物技术领域,具体涉及LncRNA-ENST00000446135在制备预测AML预后试剂盒中的应用。
背景技术
急性髓性白血病(AML)是一组具有各种遗传异常的造血系统恶性肿瘤,包括染色体易位和/或体细胞突变,其主要造成骨髓祖细胞的异常增殖、分化或存活。尽管多种治疗方法如化疗和干细胞移植等常用于AML的治疗,但其预后仍不容乐观,尤其是化疗耐药、转移及复发的产生对预后不良产生不容忽视的重要作用。目前多项研究表明,表观遗传失调对AML的作用越来越受重视,除了乙酰化及甲基化修饰,非编码RNA的调节也发挥重要作用。
已有研究表明,LncRNA在分子遗传水平和细胞水平具有相应的功能,包括参与基因组印记、剂量补偿效应、染色质的修饰、细胞分化、细胞周期的调控、细胞内物质的运输以及热休克反应等。如HOXA-AS2在体外和体内能够促进ADR细胞增殖并抑制细胞凋亡导致AML的发生,同时其能负调节ADR细胞中miR-520c-3p的表达来增强AML的化疗药物耐药。LncRNAANRIL通过AdipoR1/AMPK/SIRT1的葡萄糖代谢途径促进恶性细胞存活和细胞葡萄糖代谢以加速AML进展,其可作为AML治疗中潜在的预后标志物和治疗靶标。然而目前用于AML预后预测的标志物对AML预后指示的准确度不高,因此,开发一种具有更优指示AML预后的标志物对于AML更准确的预后具有重要意义。
发明内容
针对现有技术存在的问题,本发明目的在于提供LncRNA-ENST00000446135在制备预测AML预后试剂盒中的应用,本发明以LncRNA-ENST00000446135作为AML预后预测标志物,对于AML的预后预测具有较高的准确度。
基于上述目的,本发明采用的技术方案如下:
第一方面,本发明提供LncRNA-ENST00000446135在制备急性髓性白血病预后预测试剂盒中的应用。
LncRNA-ENST00000446135的核苷序列如SEQ ID NO.1所示,本发明通过对正常细胞和AML细胞的RNA-seq的差异性分析发现在AML细胞中LncRNA-ENST00000446135的表达水平显著低于正常细胞,并且在AML患者的标本中得到验证,LncRNA-ENST00000446135在AML初诊及复发患者中异常低表达,但在化疗缓解及造血干细胞移植后该LncRNA能恢复正常表达,表明LncRNA-ENST00000446135是AML患者生存时间的独立影响因素,因此,LncRNA-ENST00000446135可作为AML患者预后预测标志物。
优选地,通过检测临床患者的骨髓中LncRNA-ENST00000446135的表达水平,结合判定标准,从而预测AML预后。
优选地,上述判定标准如下:
当LncRNA-ENST00000446135的表达水平CT值≥0.655时,指示AML患者预后良好;
当LncRNA-ENST0000044613的表达水平CT值<0.655时,指示AML患者预后不良。
基于本发明判定标准,能够更为准确指示AML患者的预后。
第二方面,本发明提供一种用于急性髓性白血病预后预测试剂盒,所述试剂盒包括用于扩增LncRNA-ENST00000446135的引物;所述引物如下:
LncRNA-ENST00000446135(F):5'-GGGACAAGCAGCACAGAACT-3'(SEQ ID NO.2);
LncRNA-ENST00000446135(R):5'-CAGCAGAATAACGGCACAAG-3'(SEQ ID NO.3)。
优选地,所述的试剂盒还包括用于扩增内参的引物,具体如下:
18SrRNA(F):5'-CGGCGGCTTTGGTGACTCTAGA-3'(SEQ ID NO.4);
18SrRNA(R):5'-CCTGCTGCCTTCCTTGGATGTG-3'(SEQ ID NO.5)。
优选地,所述试剂盒还包括用于分离骨髓单个核细胞的试剂、用于总RNA提取的试剂、用于逆转录PCR的试剂、用于实时定量PCR的试剂中的任意一种或至少两种。
第三方面,本发明提供上述用于急性髓性白血病预后预测试剂盒在非诊断检测LncRNA-ENST00000446135表达中应用。
与现有技术相比,本发明的有益效果如下:
本发明通过对正常细胞和AML细胞的RNA-seq的差异性分析发现在AML细胞中LncRNA-ENST00000446135的表达水平显著低于正常细胞,并且在AML患者的标本中得到验证,LncRNA-ENST00000446135在AML初诊及复发患者中异常低表达,但在化疗缓解及造血干细胞移植后该LncRNA能恢复正常表达,表明LncRNA-ENST00000446135是AML患者生存时间的独立影响因素,提示该LncRNA作为AML预后预测的标志物对于AML的预后预测具有较高的准确度,为AML的诊断、危险分层及预后提供更多信息,也为AML的靶向治疗奠定基础。
附图说明
图1为实施例1所述病患的生存曲线;
图2为过表达LncRNAENST00000446135的THP-1和U937细胞株凋亡实验的流式结果图;
图3为过表达LncRNA-ENST00000446135的THP-1和U937细胞株的细胞活性图。
具体实施方式
为更好地说明本发明的目的、技术方案和优点,下面将结合具体实施例对本发明作进一步说明。本领域技术人员应当理解,此处所描述的具体实施例仅用以解释本发明,并不用于限定本发明。
实施例中所用的试验方法如无特殊说明,均为常规方法;所用的材料、试剂等,如无特殊说明,均可从商业途径得到。其中,淋巴细胞分离液Ficoll购自GE Healthcare LifeSciences;TRIZOL购自Invitrogen;逆转录PCR试剂盒购自Promega;实时定量PCR试剂盒购自TIANGEN;细胞MTS增殖检测试剂盒购自Promega。
实施例1
1、样本收集
在与患者签署知情同意书的前提下采骨髓。收集2014年11月至2018年12月在广州市第一人民医院的95名AML患者以及12名健康成人的骨髓标本及临床资料,其中47例为初诊AML患者,7例为复发AML患者,24例为化疗后AML患者,17例为造血干细胞移植后的AML患者。根据国家综合癌症网络(NCCN)急性髓性白血病指南的细胞遗传学和分子异常进行危险分层。由于只有3名初诊AML的患者为中危,因此将患者分为高/中危(PR/IM risk)和低危(Favorable risk)两组。所有标本取自于入院时的骨髓肝素抗凝,该部分研究方案已经获得本单位伦理委员会通过。同时收集AML患者生存时间和生存状态等临床资料(如表1所示)。
2、人骨髓单个核细胞提取方法
取上述正常人、AML患者的骨髓(肝素抗凝)及外周血标本2~5ml(血常规管);在15ml离心管中加入5ml淋巴细胞分离液;取标本与等量RPMI1640充分混匀,用滴管沿管壁缓慢叠加于Ficoll(1.077g/ml,GE公司)分层液面上,注意保持清楚的界面。水平离心2000rpm×20分钟;离心后管内分为三层,上层为血浆和培养液,下层主要为红细胞和粒细胞。中层为淋巴细胞分离液,在上、中层界面处有一以单个核细胞为主的白色云雾层狭窄带,单个核细胞包括淋巴细胞和单核细胞;用毛细血管插到云雾层,吸取单个核细胞。置入另一离心管中,加入5倍以上体积的RPMI1640,1500rpm×5分钟,洗涤细胞两次;末次离心后,弃上清,加入RPMI1640,重悬细胞。
适当稀释,于血球计数板上,计数四个大方格内的细胞总数。单个核细胞浓度(细胞数/1毫升细胞悬液)=4个大方格内细胞总数/4×104×(稀释倍数);
取离心管细胞悬液离心沉淀单个核细胞,弃上清,加入1ml Trizol(Invitrogen)溶液,混匀后置1.5ml EP管中,冻存于-80℃冰箱,用于后续进行RNA提取等。
3、总RNA提取
将单细胞悬液置于冰上,5分钟后,加入氯仿0.2毫升,充分混匀(15秒)后置冰上2-3分钟,低温高速(2-8℃,12000g)离心15-30分钟;
小心吸取上层液体(约占总体积50%)移至1.5毫升新管中,加入0.5毫升异丙醇并混匀,冰上放置10分钟后低温高速(2-8℃,12000g)离心10-20分钟;
去上清并用1毫升75%乙醇(-20℃)洗涤两次,每次均混匀30秒后,(2-8℃,10000g)离心5-10分钟;
去上清,再离心一次,去除剩余的乙醇;
于低温2~8℃真空离心机中干燥5-10分钟;
加入超净水(Biotecx BL-5700)50微升(视情况可适当增加或减少)混匀,得到RNA样品。
取2微升RNA样品用超净水稀释至400微升,于紫外线分光光度仪中检测样品在A260nm波长的光密度,估计样品的纯度和含量(1ODA260nm=40微克/毫升),RNA总量=OD数×400微克;
RNA样品加入0.1倍容积(5微升)的3mol/L NaAC(醋酸钠)和2倍容积(100微升)的乙醇后,保存于-70℃备用。
4、RT-PCR
根据总RNA含量及所需cDNA量来计算加样体系,以加样总RNA量为500ng为准,1000ng/RNA浓度=RNA体积;
取出逆转录试剂盒置于冰上,按GoScript TM RT-PCR试剂盒说明书配制逆转录体系(20μl)如下:RNA500ng,Olig(dT)(0.5μg/reaction)0.5μl,Random primer(0.5μg/reaction)0.5μl,加入RNase free ddH2O至5μl,70℃5min后迅速冰上冷却至少5min;
然后加入GoScript TM 5×Reaction Buffer 4.0μl、MgCl2(终浓度1.5-5.0mM)1.7μl、0.5mM dNTP Mix 1.0μl、Ribonuclease inhibitor(20units)0.3μl、ReverseTranscriptase 1.0μl,加入ddH2O至15μl,混匀后42℃延伸60min,70℃灭活15min,最后-20℃保存备用。
5、qRT-PCR
其中,所用引物的序列如下:
LncRNA-ENST00000446135(F):5’-GGGACAAGCAGCACAGAACT-3’(SEQ ID NO.2);
LncRNA-ENST00000446135(R):5’-CAGCAGAATAACGGCACAAG-3’(SEQ ID NO.3);
内参引物:
18SrRNA(F):5'-CGGCGGCTTTGGTGACTCTAGA-3'(SEQ ID NO.4);
18SrRNA(R):5'-CCTGCTGCCTTCCTTGGATGTG-3'(SEQ ID NO.5);
用ddH2O将cDNA稀释10倍,引物稀释10倍,按GoTaq@qPCR Master Mix试剂盒配制PCR反应体系(10ul)如下:GoTaq@Master Mix 2X 5μl,上游引物、下游引物(10μM)各0.5μl,cDNA2μl(cDNA稀释10倍),ddH2O 2μl。
将以上各试剂配制成预混体系后离心混匀,将各样品体系加到96孔板或者八联管,每个样品设置三个复孔;
使用ViiA7 TM System software软件设定反应程序:95℃10min;95℃15S,60℃1min,40个循环。
运行结束后查看各组基因扩增曲线和溶解曲线,读取分析各样品Tm值和Ct值,Ct值用excel软件处理,采用归一法对各组样品中目标基因的相对表达量进行统计分析。
对qRT-PCR测定得到的基因表达量CTLncRNA-ENST00000446135,进一步采用ΔΔCT法进行统计学分析,得到基因表达水平,其中:
ΔCTLncRNA-ENST00000446135=(CTLncRNA-ENST00000446135-CT18SrRNA);
ΔΔCT=ΔCT初发AML患者–ΔCT健康成人;步骤(1)中健康成人骨髓样本有12例,ΔCT健康成人是通过计算平均值得到的;基因表达水平=2^(-ΔΔCT)。基因的表达情况以及AML患者临床预后资料如图1(对LncRNA-ENST00000446135进行高低表达分组后的随访生存时间)及下表1所示。
表1
从上述测得的基因表达水平虽然不能直接得出将来诊断结果和健康状况,但其作为中间结果,可以作为患者临床治疗方案制定的参考信息之一。
6、过表达LncRNA-ENST00000446135的细胞凋亡实验
6.1慢病毒包装
(1)慢病毒包装流程
制备慢病毒穿梭质粒及其辅助包装原件载体质粒,三种质粒载体分别进行高纯度无内毒素抽提,共转染293T细胞,转染后6h更换为完全培养基,培养48h和72h后,收集富含慢病毒颗粒的细胞上清液,4℃,2000×g,10min,去除细胞碎片,然后收集病毒上清液,利用超离:4℃,82700×g,离心120min,对其超离,最后得到高滴度的慢病毒超离液。
(2)慢病毒载体、包装细胞和菌株
病毒包装系统:三质粒系统,pSPAX2、pMD2G和穿梭质粒(携带目的基因或者shRNA)。包装细胞株:293T,慢病毒的包装细胞,为贴壁依赖型成上皮样细胞,生长培养基为DMEM(含10%FBS)。贴壁细胞经培养生长增殖形成单层细胞。菌株:大肠杆菌菌株DH5-α,用于扩增慢病毒载体和辅助包装载体质粒。
(3)质粒扩增
构建好的慢病毒载体和辅助质粒需要大量抽提,浓度大于1μg/μL,A260/280在1.7~1.8之间方可用以包毒。
(4)病毒包装
第一天:铺板293T细胞用于转染(前提是细胞已经培养到传代后可以满足后续转染实验需要)。操作完毕后置于37℃,5% CO2和95%相对湿度的培养箱中。
第三天:转染
①观察细胞密度,达到70~80%的汇合率即可进行转染。
②做脂转complex:Opti MEM需在37℃水浴中预热,Lipofiter TM转染试剂需恢复至室温方可使用,使用前需摇匀。
③转染后更换含10%胎牛血清FBS的新鲜完全培养基,如果在当天上午进行转染,转染后6h进行换液,如果当天下午进行转染,转染后第二天早上约转染后16h进行换液。
④收毒:转染后48h和72h分别两次收集病毒上清(48h收集后置换新鲜完全培养基)。在48h收毒时,将100mm dish中的培养基倒入50mL离心管中,注意培养皿壁不要接触离心管口,以防出现细菌污染,随后补入10mL含10%胎牛血清FBS的新鲜完全培养基,平稳置于37℃,5% CO2的恒温培养箱中继续培养。在72h收毒时,直接将100mm dish中的培养基倒入50mL离心管中,同样注意培养皿壁不要接触离心管口,以防出现细菌污染。收完72h的病毒原液后,该dish便可以舍弃。
⑤超速离心:将50mL离心管中的病毒上清,4℃,2000×g,离心10min,去除细胞碎片;然后收集病毒原液上清置于超速离心管中,4℃,82700×g,离心120min,最后将慢病毒超离液分装到灭菌处理的病毒管中。
⑥病毒保存:按照要求分装病毒,标记(病毒名称,年-月-日),-80℃冰箱保存
(5)Lipofiter TM脂质体转染试剂操作指南
①细胞培养(以十二孔板为例):在转染前一天(20-24h)把约0.5-40万细胞(具体的细胞数量据细胞大小和细胞生长速度而定)培养到十二孔板内,使第二天细胞能达到约70-80%。
②在进行下述转染步骤前,把十二孔板每孔内换成新鲜的细胞培养液。培养液的体积约为1mL左右,可以含有血清和抗生素。
③把Lipofiter TM脂质体转染试剂轻轻混匀。
④对于待转染的十二孔板中一个孔的细胞,取一只洁净无菌离心管,加入1.6μgDNA及适量的不含抗生素和glutamine的DMEM溶液,用枪轻轻吹打混匀,终体积100μL。
⑤取另一只洁净无菌离心管,加入95.2μL不含抗生素和glutamine的DMEM溶液,再加入4.8μL Lipofiter TM,用枪轻轻吹打混匀,终体积100μL,室温放置5min。
⑥将步骤4与5中的DNA溶液与Lipofiter TM溶液混合,用枪轻轻吹打混匀。注:不可Vortex或离心。
⑦室温孵育20min。有可能出现絮状沉淀物,属正常现象,不会影响转染效率。⑧无论是贴壁细胞还是悬浮细胞,均把200μL Lipofiter TM-DNA混合物全部加入十二孔板的一个孔内。加入时注意尽量均匀加入到整个孔内,随后轻轻8字摇摆混匀。
⑨细胞培养箱内培养6h后,去除含有Lipofiter TM-DNA的培养液。每孔加入1mL新鲜细胞培养液继续培养。
注:通常Lipofiter TM-DNA混合物和细胞一起孵育3-6h已经足够产生较高的转染效率。大多数细胞和Lipofiter TM-DNA一起培养长达72h未见明显细胞毒性。
⑩转染后续处理
1)对于基因表达,再培养24-40h后即可检测转染效果,如转染带GFP或者其他荧光基因的表达载体,可用荧光显微镜观测细胞转染效率。
2)对于siRNA(合成的小RNA),一般在24-72h检测下调效率,期间需要更换培养液。对于干扰质粒载体,一般在36-96h检测下调效率。
3)用于筛选稳定表达细胞株,则在转染后24h即可加入适当的筛选药物,例如G418或嘌呤霉素等,进行稳定表达细胞株的筛选。
(6)滴度检测
(1)细胞准备
将生长状态良好的293T细胞消化计数后稀释至1×105/mL,加入96孔板,100μL/孔,为每个病毒准备6个孔。放入37℃,5% CO2培养箱中培养。
(2)加病毒,第二天,准备6个1.5mL EP管,第一个EP管中加入10μL病毒液,然后做3倍梯度稀释,共6个稀释度。
(3)追加培养液,第三天,有需要加puromycin筛选的孔,先吸去100mL含病毒培养基,加入100μL含1.5μg/mL puromycin的10% FBS完全培养基。
(4)观察结果并计算滴度
(5)第五天,在荧光显微镜下观察结果,在观察结果前6h需更换新鲜10%FBS完全培养基,从孔中吸出80μL培养基,然后加入80μL新鲜10%FBS完全培养基,放入37℃,5%CO2培养箱中培养。6h后荧光显微镜下观察结果,荧光百分比在10~50%的孔计算病毒滴度。滴度(TU/mL)=细胞数×阳性克隆百分比×MOI(1)×病毒稀释倍数×103TU/mL
2.6.4感染细胞最佳MOI(感染指数)的测定
(1)取生长状态良好的THP-1及U937,分别以3×104个/孔的密度在24孔培养板中常规培养24h,细胞经过24h的培育后每孔数目大约为6×104个/孔;
(2)接种24h后吸走500μl培养基,根据病毒数加入慢病毒;
(3)从-80℃冰箱取出病毒,于冰上溶解,于离心机中离心后备用;
(4)病毒感染:①取THP-1及U937细胞以MOI分别为20、30、40、50、60、70、80、90、100,加入慢病毒及polybrene 3μg/ml×500μl即1.5μg 0.75μl;
(5)感染24h后加培养基至1ml;
(6)感染48h后在荧光显微镜和流式下观测荧光表达情况,依据细胞感染效果,确定THP-1最佳MOI=70,U937最佳MOI=40。
2.6.5杀伤曲线的检测
(1)分别把THP-1及U937以1×104个/孔的密度铺于96孔板;
(2)24h后,加入不同浓度的嘌呤霉素,浓度依次为0、0.2ug/ml、0.4ug/ml、0.6μg/ml、0.8μg/ml、1.0μg/ml、1.2μg/ml、1.4μg/ml、1.6μg/ml,每个浓度6个平行孔;
(3)连续观察一周,中间换依次液,选择细胞刚好完全死亡的孔的加药浓度为筛选浓度,即THP-1及U937的筛选浓度均为1μg/ml。
2.6.6慢病毒转染细胞步骤
(1)取生长状态良好的THP-1及U937,分别以3×104个/孔的密度在24孔培养板中常规培养24h,细胞经过24h的培育后每孔数目大约为6×104个/孔;
(2)接种24h后吸走500μl培养基,根据病毒数加入慢病毒;
(3)从-80℃冰箱取出病毒,于冰上溶解,于离心机中离心后备用;
(4)病毒感染:①取THP-1细胞的MOI=70,加入慢病毒数4.2×106个(42μl);②取U937细胞的MOI=40,加入慢病毒数2.4×106个(24μl);③加入polybrene3μg/ml×500μl即1.5μg 0.75μl;
(5)感染24h后加培养基至1ml;
(6)感染48h后换成带有浓度为1μg/ml嘌呤霉素的完全培养基进行细胞筛选,每2天进行换液,同时以空白细胞作为对照;
(7)等空白组的对照细胞全部死亡时,换成0.5μg/ml的嘌呤霉素完全培养基培养以抑制基因外排;
(8)等细胞长满后进行扩大培养,并取部分细胞提取基因进行qRT-PCR实验与空白细胞进行对比;
(9)得到的稳定转染株分别为:THP-1过表达细胞(Lenti-lncRNA-OV)、U937过表达细胞(Lenti-lncRNA-OV)、THP-1阴性对照细胞(NC)、U937阴性对照细胞(NC)。
6.1、细胞收集及染色
用PBS洗涤细胞二次(2000rpm离心5min)收集1~5×105细胞;加入500μl的Binding Buffer悬浮细胞;加入5μl Annexin V-APC混匀后,加入5μl Propidium Iodide,混匀;室温、避光、反应5~15min;在1h内,进行下述荧光显微镜或流式细胞仪的观察和检测。
6.2流式细胞仪分析
用流式细胞仪检测,激发波长Ex=488nm;发射波长Em=530nm;
激发波长633nm,最大发射波长660nm,Annexin V-APC的红色荧光建议使用;
FL4通道检测;PI红色荧光(流式Ex=488nm,Em≥630nm)通过FL2或FL3通道检测,建议使用FL3;
荧光补偿调节:使用经凋亡诱导处理的正常细胞,作为对照进行荧光补偿调节去除光谱重叠和设定十字门的位置。
结果如图2所示,流式结果显示,U937和THP-1过表达的细胞株较空白和对照组,凋亡均有明显增加。
7、过表达lncRNA的细胞增殖实验
分别培养THP-1、THP-1(NC)、THP-1(Lenti-lncRNA-OV);U937、U937(NC)、U937(Lenti-lncRNA-OV)细胞于25cm 2培养瓶至80%-90%覆盖度;其中,THP-1指未做处理的野生型细胞;THP-1(Lenti-lncRNA-OV)指的是对THP-1细胞株做LncRNA-ENST00000446135过表达的处理;THP-1(NC)指LncRNA-ENST00000446135过表达对照细胞。U937指未做处理的野生型细胞;U937(Lenti-lncRNA-OV)指对U937细胞株做LncRNA-ENST00000446135过表达的处理;U937(NC)指LncRNA-ENST00000446135过表达对照细胞。
收集以上各细胞,计数后RPMI1640完全培养基制成单细胞悬液,按10000/孔种96孔板,每孔体积100μl;
培养细胞一定时间,使其在不同的时间点(24h/48h/72h/96h)向每孔加入20μlMTS溶液,37℃、5%CO2的培养箱孵育1~4小时;(注:如果需要以后检测,每孔加入25μl10%SDS终止反应,避光保存于室温的湿盒中,最多可保存18h)使用酶标仪选择490nm处波长,检测96孔板的吸光度值。统计不同时间点各细胞的吸光度,对比各细胞在不同时间点增殖的差异,结果如图3所示,u937和THP-1过表达的细胞株较空白和对照组,增殖能力均有明显降低。证明LncRNA-ENST00000446135在AML肿瘤细胞的增殖有着调控作用。
最后所应当说明的是,以上实施例仅用以说明本发明的技术方案而非对本发明保护范围的限制,尽管参照较佳实施例对本发明作了详细说明,本领域的普通技术人员应当理解,可以对本发明的技术方案进行修改或者等同替换,而不脱离本发明技术方案的实质和范围。
Claims (7)
1.LncRNA-ENST00000446135在制备急性髓性白血病预后预测试剂盒中的应用。
2.如权利要求1所述LncRNA-ENST00000446135在制备急性髓性白血病预后预测试剂盒中的应用,其特征在于,通过检测临床患者的骨髓中LncRNA-ENST00000446135的表达水平,结合判定标准,从而预测AML预后。
3.如权利要求1所述LncRNA-ENST00000446135在制备急性髓性白血病预后预测试剂盒中的应用,其特征在于,所述判定标准如下:
当LncRNA-ENST00000446135的表达水平CT值≥0.655时,指示AML患者预后良好;
当LncRNA-ENST0000044613的表达水平CT值<0.655时,指示AML患者预后不良。
4.一种用于急性髓性白血病预后预测试剂盒,其特征在于,所述试剂盒包括用于扩增LncRNA-ENST00000446135的引物;所述引物如下:
LncRNA-ENST00000446135(F):5'-GGGACAAGCAGCACAGAACT-3';
LncRNA-ENST00000446135(R):5'-CAGCAGAATAACGGCACAAG-3'。
5.如权利要求4所述用于急性髓性白血病预后预测试剂盒,其特征在于,所述的试剂盒还包括用于扩增内参的引物,具体如下:
18SrRNA(F):5'-CGGCGGCTTTGGTGACTCTAGA-3';
18SrRNA(R):5'-CCTGCTGCCTTCCTTGGATGTG-3'。
6.如权利要求4所述用于急性髓性白血病预后预测试剂盒,其特征在于,所述试剂盒还包括用于分离骨髓单个核细胞的试剂、用于总RNA提取的试剂、用于逆转录PCR的试剂、用于实时定量PCR的试剂中的任意一种或至少两种。
7.权利要求4~6任一项所述用于急性髓性白血病预后预测试剂盒在非诊断检测LncRNA-ENST00000446135表达中应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202211743017.7A CN116103400B (zh) | 2022-12-30 | 2022-12-30 | LncRNA-ENST00000446135在制备预测AML预后试剂盒中的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202211743017.7A CN116103400B (zh) | 2022-12-30 | 2022-12-30 | LncRNA-ENST00000446135在制备预测AML预后试剂盒中的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN116103400A true CN116103400A (zh) | 2023-05-12 |
CN116103400B CN116103400B (zh) | 2023-12-01 |
Family
ID=86255563
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202211743017.7A Active CN116103400B (zh) | 2022-12-30 | 2022-12-30 | LncRNA-ENST00000446135在制备预测AML预后试剂盒中的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN116103400B (zh) |
Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113151470A (zh) * | 2021-04-26 | 2021-07-23 | 暨南大学 | 多基因联合在制备预测aml预后试剂盒中的应用 |
CN113549679A (zh) * | 2021-07-08 | 2021-10-26 | 南京市儿童医院 | LncRNA ANRIL在儿童急性淋巴细胞白血病中的临床应用 |
-
2022
- 2022-12-30 CN CN202211743017.7A patent/CN116103400B/zh active Active
Patent Citations (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113151470A (zh) * | 2021-04-26 | 2021-07-23 | 暨南大学 | 多基因联合在制备预测aml预后试剂盒中的应用 |
CN113549679A (zh) * | 2021-07-08 | 2021-10-26 | 南京市儿童医院 | LncRNA ANRIL在儿童急性淋巴细胞白血病中的临床应用 |
Non-Patent Citations (2)
Title |
---|
ZHIHENG ZHOU等: "Long non-coding RNAs as novel expression signatures modulate DNA damage and repair in cadmium toxicology", SCIENTIFIC REPORTS, vol. 5, pages 10 - 11 * |
王彩霞等: "长链非编码RNA对急性髓系白血病细胞增殖和凋亡调控研究", 中华肿瘤防治杂志, vol. 25, no. 3, pages 159 - 164 * |
Also Published As
Publication number | Publication date |
---|---|
CN116103400B (zh) | 2023-12-01 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
Che et al. | RETRACTED: Exosomes Derived from miR-143-Overexpressing MSCs Inhibit Cell Migration and Invasion in Human Prostate Cancer by Downregulating TFF3 | |
Horos et al. | Ribosomal deficiencies in Diamond-Blackfan anemia impair translation of transcripts essential for differentiation of murine and human erythroblasts | |
Zhang et al. | LncRNA SNHG14 promotes the development of cervical cancer and predicts poor prognosis. | |
CN104685356A (zh) | 通过过滤从生物样品提取或分离的罕见细胞的多种分析方法 | |
Deng et al. | Aberrant NEAT1_1 expression may be a predictive marker of poor prognosis in diffuse large B cell lymphoma | |
CN111218515A (zh) | 多组织器官和细胞类型的衰老标记及卡路里限制在延缓机体衰老中的应用 | |
Marcovecchio et al. | Premature senescence and increased oxidative stress in the thymus of down syndrome patients | |
CN109908369B (zh) | 一种新的环状RNA circCRKL在前列腺癌治疗中的应用 | |
Zhao et al. | Acute myeloid leukemia cell-derived extracellular vesicles carrying microRNA-548ac regulate hematopoietic function via the TRIM28/STAT3 pathway | |
CN104208723B (zh) | miR-638在抗急性髓系白血病中的应用 | |
Liu et al. | LncRNA VPS9D1-AS1 promotes malignant progression of lung adenocarcinoma by targeting miRNA-30a-5p/KIF11 axis | |
Xu et al. | Circular RNA circSIPA1L1 contributes to osteosarcoma progression through the miR-411-5p/RAB9A signaling pathway | |
Shen et al. | Downregulation of UBE2T can enhance the radiosensitivity of osteosarcoma in vitro and in vivo | |
CN116103400B (zh) | LncRNA-ENST00000446135在制备预测AML预后试剂盒中的应用 | |
CN113908283A (zh) | Prmt5抑制剂及其与pd-l1抗体阻断剂联合在治疗肺癌上的应用 | |
Fang et al. | PUS1 is a novel biomarker for predicting poor outcomes and triple-negative status in breast cancer | |
Wang et al. | Role of methylation-related genes CRYAB and SLC39A11 in the occurrence and development of lung adenocarcinoma | |
CN106148337B (zh) | 长非编码rna ay927503及其用途 | |
Che et al. | RETRACTED: MicroRNA-27 Inhibits Autophagy and Promotes Proliferation of Multiple Myeloma Cells by Targeting the NEDD4/Notch1 Axis | |
CN114703190B (zh) | 一种靶向抑制KIAA1429基因表达的ShRNA在慢性髓细胞白血病中的应用 | |
Yang et al. | Retracted Paper-Downregulation of microRNA-586 Inhibits Proliferation, Invasion and Metastasis and Promotes Apoptosis in Human Osteosarcoma U2-OS Cell Line | |
Maifrede et al. | Loss of Egr1, a human del5q gene, accelerates BCR-ABL driven chronic myelogenous leukemia | |
Huang et al. | VPS9D1-AS1, a novel long-non-coding RNA, acts as a tumor promoter by regulating the miR-324-5p/ITGA2 axis in colon adenocarcinoma | |
CN113265428B (zh) | 一种利用金属硫蛋白构建活细胞内铜变化的检测系统及应用 | |
CN110157736B (zh) | 一种促进山羊毛囊干细胞增殖的方法 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |