CN116286828B - Oligonucleotide siRNA and application thereof in preparation of drugs for preventing and treating liver cancer - Google Patents
Oligonucleotide siRNA and application thereof in preparation of drugs for preventing and treating liver cancer Download PDFInfo
- Publication number
- CN116286828B CN116286828B CN202310531654.6A CN202310531654A CN116286828B CN 116286828 B CN116286828 B CN 116286828B CN 202310531654 A CN202310531654 A CN 202310531654A CN 116286828 B CN116286828 B CN 116286828B
- Authority
- CN
- China
- Prior art keywords
- liver cancer
- sirna
- application
- cancer cells
- oligonucleotide sirna
- 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.)
- Active
Links
- 201000007270 liver cancer Diseases 0.000 title claims abstract description 77
- 208000014018 liver neoplasm Diseases 0.000 title claims abstract description 76
- 108091034117 Oligonucleotide Proteins 0.000 title claims abstract description 36
- 239000003814 drug Substances 0.000 title claims abstract description 19
- 108020004459 Small interfering RNA Proteins 0.000 title abstract description 40
- 229940079593 drug Drugs 0.000 title abstract description 4
- 238000002360 preparation method Methods 0.000 title description 4
- 230000012010 growth Effects 0.000 claims abstract description 14
- 230000005012 migration Effects 0.000 claims abstract description 14
- 238000013508 migration Methods 0.000 claims abstract description 14
- 108020004707 nucleic acids Proteins 0.000 claims abstract description 12
- 102000039446 nucleic acids Human genes 0.000 claims abstract description 12
- 150000007523 nucleic acids Chemical class 0.000 claims abstract description 12
- 230000035755 proliferation Effects 0.000 claims abstract description 9
- KZMAWJRXKGLWGS-UHFFFAOYSA-N 2-chloro-n-[4-(4-methoxyphenyl)-1,3-thiazol-2-yl]-n-(3-methoxypropyl)acetamide Chemical compound S1C(N(C(=O)CCl)CCCOC)=NC(C=2C=CC(OC)=CC=2)=C1 KZMAWJRXKGLWGS-UHFFFAOYSA-N 0.000 claims abstract description 6
- 210000004027 cell Anatomy 0.000 claims description 89
- 230000002401 inhibitory effect Effects 0.000 claims description 15
- 230000005764 inhibitory process Effects 0.000 claims description 10
- 230000015572 biosynthetic process Effects 0.000 claims description 8
- 238000004519 manufacturing process Methods 0.000 claims description 6
- 210000005229 liver cell Anatomy 0.000 claims description 2
- 230000002414 glycolytic effect Effects 0.000 abstract description 10
- 238000002474 experimental method Methods 0.000 abstract description 7
- 108091032973 (ribonucleotides)n+m Proteins 0.000 abstract description 4
- 238000011161 development Methods 0.000 abstract description 3
- 206010073071 hepatocellular carcinoma Diseases 0.000 description 25
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 14
- 102100032921 ATP-dependent 6-phosphofructokinase, liver type Human genes 0.000 description 13
- 101000730830 Homo sapiens ATP-dependent 6-phosphofructokinase, liver type Proteins 0.000 description 13
- 230000000694 effects Effects 0.000 description 9
- 108090000623 proteins and genes Proteins 0.000 description 9
- 230000002829 reductive effect Effects 0.000 description 9
- 238000001514 detection method Methods 0.000 description 8
- 230000009467 reduction Effects 0.000 description 8
- 239000004310 lactic acid Substances 0.000 description 7
- 235000014655 lactic acid Nutrition 0.000 description 7
- 230000012292 cell migration Effects 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 239000012528 membrane Substances 0.000 description 6
- 238000000034 method Methods 0.000 description 6
- 239000000243 solution Substances 0.000 description 6
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 6
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 206010028980 Neoplasm Diseases 0.000 description 4
- 239000000020 Nitrocellulose Substances 0.000 description 4
- 101150009383 PFKL gene Proteins 0.000 description 4
- 108091028664 Ribonucleotide Proteins 0.000 description 4
- 108091081021 Sense strand Proteins 0.000 description 4
- 239000006180 TBST buffer Substances 0.000 description 4
- 230000000692 anti-sense effect Effects 0.000 description 4
- 244000309466 calf Species 0.000 description 4
- 239000003085 diluting agent Substances 0.000 description 4
- 230000009368 gene silencing by RNA Effects 0.000 description 4
- 229920001220 nitrocellulos Polymers 0.000 description 4
- 102000004169 proteins and genes Human genes 0.000 description 4
- 239000002336 ribonucleotide Substances 0.000 description 4
- 210000002966 serum Anatomy 0.000 description 4
- 108010085238 Actins Proteins 0.000 description 3
- 102100034343 Integrase Human genes 0.000 description 3
- 108010092799 RNA-directed DNA polymerase Proteins 0.000 description 3
- 108091030071 RNAI Proteins 0.000 description 3
- 230000006536 aerobic glycolysis Effects 0.000 description 3
- 239000002299 complementary DNA Substances 0.000 description 3
- 238000007865 diluting Methods 0.000 description 3
- 230000034659 glycolysis Effects 0.000 description 3
- 231100000844 hepatocellular carcinoma Toxicity 0.000 description 3
- 108020004999 messenger RNA Proteins 0.000 description 3
- 210000004881 tumor cell Anatomy 0.000 description 3
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 2
- 239000012098 Lipofectamine RNAiMAX Substances 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 230000006682 Warburg effect Effects 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 230000005757 colony formation Effects 0.000 description 2
- 239000008367 deionised water Substances 0.000 description 2
- 229910021641 deionized water Inorganic materials 0.000 description 2
- 239000002773 nucleotide Substances 0.000 description 2
- 125000003729 nucleotide group Chemical group 0.000 description 2
- 239000000843 powder Substances 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 235000020183 skimmed milk Nutrition 0.000 description 2
- 239000012096 transfection reagent Substances 0.000 description 2
- 102000007469 Actins Human genes 0.000 description 1
- 229930024421 Adenine Natural products 0.000 description 1
- 241000283707 Capra Species 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 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
- UYTPUPDQBNUYGX-UHFFFAOYSA-N Guanine Natural products O=C1NC(N)=NC2=C1N=CN2 UYTPUPDQBNUYGX-UHFFFAOYSA-N 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 206010061309 Neoplasm progression Diseases 0.000 description 1
- 101710163270 Nuclease Proteins 0.000 description 1
- 241000283973 Oryctolagus cuniculus Species 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 102000001105 Phosphofructokinases Human genes 0.000 description 1
- 108010069341 Phosphofructokinases Proteins 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 238000012228 RNA interference-mediated gene silencing Methods 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Natural products O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 1
- JLCPHMBAVCMARE-UHFFFAOYSA-N [3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[3-[[3-[[3-[[3-[[3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-[[5-(2-amino-6-oxo-1H-purin-9-yl)-3-hydroxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxyoxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(5-methyl-2,4-dioxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(6-aminopurin-9-yl)oxolan-2-yl]methoxy-hydroxyphosphoryl]oxy-5-(4-amino-2-oxopyrimidin-1-yl)oxolan-2-yl]methyl [5-(6-aminopurin-9-yl)-2-(hydroxymethyl)oxolan-3-yl] hydrogen phosphate Polymers Cc1cn(C2CC(OP(O)(=O)OCC3OC(CC3OP(O)(=O)OCC3OC(CC3O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c3nc(N)[nH]c4=O)C(COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3COP(O)(=O)OC3CC(OC3CO)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3ccc(N)nc3=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cc(C)c(=O)[nH]c3=O)n3cc(C)c(=O)[nH]c3=O)n3ccc(N)nc3=O)n3cc(C)c(=O)[nH]c3=O)n3cnc4c3nc(N)[nH]c4=O)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)n3cnc4c(N)ncnc34)O2)c(=O)[nH]c1=O JLCPHMBAVCMARE-UHFFFAOYSA-N 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 229960000643 adenine Drugs 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000007853 buffer solution Substances 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 238000010609 cell counting kit-8 assay Methods 0.000 description 1
- 230000007910 cell fusion Effects 0.000 description 1
- 230000004663 cell proliferation Effects 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000010293 colony formation assay Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000013078 crystal Substances 0.000 description 1
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Natural products NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 1
- 229940104302 cytosine Drugs 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 230000008995 epigenetic change Effects 0.000 description 1
- 235000013861 fat-free Nutrition 0.000 description 1
- 230000030279 gene silencing Effects 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 230000002068 genetic effect Effects 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 230000035876 healing Effects 0.000 description 1
- 238000003119 immunoblot Methods 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000006371 metabolic abnormality Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 230000017066 negative regulation of growth Effects 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 230000010627 oxidative phosphorylation Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 230000037361 pathway Effects 0.000 description 1
- 238000003752 polymerase chain reaction Methods 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 238000010837 poor prognosis Methods 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 239000004055 small Interfering RNA Substances 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 238000003892 spreading Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000001890 transfection Methods 0.000 description 1
- 238000003151 transfection method Methods 0.000 description 1
- 238000012546 transfer Methods 0.000 description 1
- 239000000107 tumor biomarker Substances 0.000 description 1
- 230000004614 tumor growth Effects 0.000 description 1
- 230000005751 tumor progression Effects 0.000 description 1
- 229940035893 uracil Drugs 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N15/00—Mutation or genetic engineering; DNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purification; Use of hosts therefor
- C12N15/09—Recombinant DNA-technology
- C12N15/11—DNA or RNA fragments; Modified forms thereof; Non-coding nucleic acids having a biological activity
- C12N15/113—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing
- C12N15/1135—Non-coding nucleic acids modulating the expression of genes, e.g. antisense oligonucleotides; Antisense DNA or RNA; Triplex- forming oligonucleotides; Catalytic nucleic acids, e.g. ribozymes; Nucleic acids used in co-suppression or gene silencing against oncogenes or tumor suppressor genes
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/7088—Compounds having three or more nucleosides or nucleotides
- A61K31/713—Double-stranded nucleic acids or oligonucleotides
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P35/00—Antineoplastic agents
- A61P35/04—Antineoplastic agents specific for metastasis
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2310/00—Structure or type of the nucleic acid
- C12N2310/10—Type of nucleic acid
- C12N2310/14—Type of nucleic acid interfering N.A.
- C12N2310/141—MicroRNAs, miRNAs
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N2320/00—Applications; Uses
- C12N2320/30—Special therapeutic applications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The application discloses an oligonucleotide siRNA and application thereof in preparing a medicament for preventing and treating liver cancer. The oligonucleotide siRNA is oligonucleotide siRNA-1; the oligonucleotide siRNA-1 consists of a single-stranded nucleic acid molecule 1 and a single-stranded nucleic acid molecule 2; the single-stranded nucleic acid molecule 1 consists of SEQ ID NO:1 and 2 dT, wherein the 2 dT is positioned at the 3' end; the single-stranded nucleic acid molecule 2 consists of SEQ ID NO:2, and a single-stranded RNA molecule shown in seq id no. Experiments prove that the oligonucleotide siRNA-1 can inhibit proliferation of liver cancer cells, inhibit growth of liver cancer cells, inhibit migration of liver cancer cells and reduce glycolytic capacity of liver cancer cells. The application provides a new medicine for treating liver cancer, and has important application value for the development of liver cancer treatment and targeted medicines thereof.
Description
Technical Field
The application belongs to the biomedical field, and in particular relates to an oligonucleotide siRNA and application thereof in preparing a medicament for preventing and treating liver cancer.
Background
Liver cell liver cancer is the most common pathological type in primary liver cancer. Poor prognosis of hepatocellular carcinoma may be associated with a variety of factors, such as lack of reliable tumor biomarkers, less knowledge of genetic and epigenetic changes in tumors, insufficient in-depth study of mechanisms, etc.
Tumor cells consume glucose at a higher rate and produce more lactic acid than normally differentiated cells, a phenomenon known as aerobic glycolysis, or the Warburg effect. The Warburg effect means that even when the oxygen supply is full, tumor cells are still mainly supplied with energy through glycolysis rather than oxidative phosphorylation, meeting their high demands for nutrients. There is growing evidence that metabolic abnormalities are one of the most typical markers of tumor tissue, playing a vital role in the development and progression of tumors, while abnormal expression of aerobic glycolysis-related molecules also plays an important regulatory role in tumor progression. Thus, aerobic glycolysis is a key pathway for energy production by cancer cells, where the molecules involved may be potential biomarkers and therapeutic targets for liver cancer.
Liver Phosphofructokinase (PFKL) is a key enzyme in glycolysis process, and can catalyze and generate higher glycolysis rate, so that energy metabolism of tumor cells is abnormal, and finally growth of tumors is promoted.
RNAi (RNA interference) can specifically eliminate or shut down the expression of specific genes, and small interfering RNA is specifically introduced into cells of mammals and humans to reduce the expression of target genes by using RNAi technology, so that the expression of target proteins is reduced, and the efficient and specific gene therapy effect can be achieved.
Disclosure of Invention
The present application aims at preventing and/or treating hepatocellular carcinoma.
The application first protects the oligonucleotide siRNA-1. The oligonucleotide siRNA-1 may consist of single stranded nucleic acid molecule 1 and single stranded nucleic acid molecule 2; the single stranded nucleic acid molecule 1 may consist of SEQ ID NO:1 and 2 dT, wherein the 2 dT is positioned at the 3' end; the single stranded nucleic acid molecule 2 may consist of SEQ ID NO:2, and a single-stranded RNA molecule shown in seq id no.
In embodiments of the application, the oligonucleotide siRNA-1 may specifically be siRNA-1.
The application also protects the application of any one of the oligonucleotide siRNA-1 in preparing a medicament for preventing liver cancer.
In the above application, the prevention of liver cancer may be expressed as at least one of inhibition of proliferation of liver cancer cells, inhibition of growth of liver cancer cells, inhibition of clone formation of liver cancer cells, inhibition of migration of liver cancer cells, reduction of glycolytic capacity of liver cancer cells, reduction of lactic acid level of liver cancer cells and/or reduction of ATP content of liver cancer cells.
The application also protects the application of any one of the oligonucleotide siRNA-1 in preparing a medicament for treating liver cancer.
In the above application, the treatment of liver cancer may be expressed as at least one of inhibiting proliferation of liver cancer cells, inhibiting growth of liver cancer cells, inhibiting clone formation of liver cancer cells, inhibiting migration of liver cancer cells, reducing glycolytic capacity of liver cancer cells, reducing lactic acid level of liver cancer cells, and/or reducing ATP content of liver cancer cells.
The application also protects the application of any one of the oligonucleotide siRNA-1 in preparing a medicament for inhibiting proliferation of liver cancer cells.
The application also protects the application of any one of the oligonucleotide siRNA-1 in preparing a medicament for inhibiting the growth of liver cancer cells.
In the above application, the inhibition of the growth of liver cancer cells may be expressed as inhibition of the clone formation of liver cancer cells.
The application also protects the application of any one of the oligonucleotide siRNA-1 in preparing a medicament for inhibiting migration of liver cancer cells.
The application also protects the application of any one of the oligonucleotide siRNA-1 in preparing a medicament for reducing glycolytic capacity of liver cancer cells.
In the above application, the reduction of glycolytic capacity of the liver cancer cell may be expressed as a reduction of lactic acid level of the liver cancer cell and/or a reduction of ATP content of the liver cancer cell.
In any of the above applications, the liver cancer may be hepatocellular carcinoma.
Experiments prove that the oligonucleotide siRNA-1 can inhibit proliferation of liver cancer cells, inhibit growth of liver cancer cells, inhibit migration of liver cancer cells and reduce glycolytic capacity of liver cancer cells. The application provides a new medicine for treating liver cancer, and has important application value for the development of liver cancer treatment and targeted medicines thereof. The application has important application value.
Drawings
FIG. 1 shows the effect of quantitative PCR detection of the inhibition effect of oligonucleotide siRNA on PFKL gene and the effect of Western blot detection of the influence of oligonucleotide siRNA on PFKL protein expression.
FIG. 2 shows the effect of oligonucleotide siRNA on human hepatoma cell proliferation.
FIG. 3 shows that siRNA-1 inhibits the clone formation of human hepatoma cell HepG2.
FIG. 4 shows that siRNA-1 inhibits migration of human hepatoma cells.
FIG. 5 shows that siRNA-1 reduces glycolytic capacity of human hepatoma cells.
Detailed Description
The following detailed description of the application is provided in connection with the accompanying drawings that are presented to illustrate the application and not to limit the scope thereof. The examples provided below are intended as guidelines for further modifications by one of ordinary skill in the art and are not to be construed as limiting the application in any way.
The experimental methods in the following examples, unless otherwise specified, are conventional methods, and are carried out according to techniques or conditions described in the literature in the field or according to the product specifications. Materials, reagents and the like used in the examples described below are commercially available unless otherwise specified.
EXAMPLE 1 preparation and screening of oligonucleotide siRNA for inhibiting the expression level of PFKL protein (Gene ID: 5211)
1. Preparation of oligonucleotide siRNA for inhibiting PFKL protein expression quantity
The sense strand and the antisense strand shown in Table 1 were synthesized from the nucleotide sequence of the PFKL gene (Genebank number: NM-001002021) by Suzhou Ji Ma gene, inc. Diluting the sense strand with deionized water to obtain sense strand diluent. Diluting the antisense strand with deionized water to obtain an antisense strand diluent. And taking the sense strand diluent and the corresponding antisense strand diluent, and carrying out annealing reaction to form the oligonucleotide.
This step produced a total of 6 oligonucleotides shown in Table 1. A, G, C and U in each oligonucleotide represent adenine ribonucleotide, guanine ribonucleotide, cytosine ribonucleotide and uracil ribonucleotide in sequence. The addition of two tails of dTdT at the 3' end directs the siRNA to form RNAi silencing complexes and also avoids degradation by nucleases.
The nucleotide sequence of NC siRNA is random (as negative control).
2. Screening of oligonucleotide siRNA for inhibiting PFKL protein expression quantity
1. Acquisition of siRNA transfected human liver cancer cell HepG2
(1) Diluting the siRNA to be tested (siRNA-1, siRNA-2, siRNA-3, siRNA-4, siRNA-5 or NC siRNA) with sterile water to obtain an siRNA solution to be tested with the concentration of 20 mu M.
(2) Digestion of human hepatoma cell HepG2 (product of American ATCC cell bank, catalog number HB-8065), spreading on a petri dish (specification 6 cm. Times.6 cm), adding DMEM medium containing 10% new born calf serum, 37℃and 5% CO 2 Culturing until the cell density reaches 50-70%.
(3) The siRNA solution to be tested obtained in step (1) and the cells obtained in step (2) were transfected with Lipofectamine RNAimax transfection reagent (Invitrogen, usa) and the transfection method was referred to the instructions of Lipofectamine RNAimax transfection reagent. And collecting cells 48h after transfection, thus obtaining the siRNA transfected human liver cancer cell HepG2.
2. Real-time quantitative PCR detection of inhibition effect of oligonucleotide siRNA on PFKL gene
(1) Total RNA of siRNA transfected human hepatoma cell HepG2 obtained in step 1 was extracted using Trizol (Invitrogen).
(2) Reverse transcription
(a) Preparing a system. The system was 15.9. Mu.l, consisting of 2. Mu.g total RNA of siRNA transfected human hepatoma cells HepG2, 1. Mu.l of 1. Mu.g/. Mu.l of random primer aqueous solution and DEPC water.
The random primer is synthesized by Beijing Saighur company.
(b) Taking the system prepared in the step (a), incubating for 5min at 70 ℃, and cooling on ice.
(c) After completion of step (b), 5. Mu. l M-MLV 5 Xbuffer, 2.5. Mu.l dNTP aqueous solution (concentration: 10 mM) and 1.0. Mu. l M-MLV reverse transcriptase (concentration: 200U/. Mu.l) were added, and the mixture was mixed, incubated at 42℃for 60min, and terminated at 95℃for 5min to obtain cDNA of siRNA-transfected human hepatoma cell HepG2.
M-MLV reverse transcriptase is a product of Promega corporation, USA. M-MLV 5 Xbuffer is a module in M-MLV reverse transcriptase.
(3) Real-time quantitative PCR (polymerase chain reaction) detection of relative expression level of PFKL (beta-actin gene serving as an internal reference gene) in cDNA (complementary deoxyribonucleic acid) of siRNA transfected human hepatoma cell HepG2, specifically adopting 2 -ΔΔCT The relative expression level of PFKL mRNA was calculated by the method.
The primers for detecting the PFKL gene are: 5'-GGAGAAGCTGCGCGAGGTTTAC-3' and 5'-ATTGTGCCAGCATCTTCAGCATGAG-3'.
Primers for detecting the beta-actin gene were 5'-TCGTGCGTGACATTAAGGAG-3' and 5'-ATGCCAGGGTACATGGTGGT-3'.
The detection results are shown in fig. 1, wherein a represents p < 0.01, and p < 0.05. The results show that the relative expression levels of PFKL mRNA of the human hepatoma cell HepG2 transfected by the siRNA-1, the siRNA-2, the siRNA-3, the siRNA-4 and the siRNA-5 are all remarkably reduced compared with the NC siRNA, wherein the reduction of the relative expression level of the PFKL mRNA of the human hepatoma cell HepG2 transfected by the siRNA-1 is most remarkable.
3. Influence of protein immunoblotting detection oligonucleotide siRNA on PFKL protein expression quantity
(1) Respectively extracting total protein of the siRNA transfected human liver cancer cell HepG2 obtained in the step 1, and carrying out SDS-PAGE electrophoresis after denaturation; the voltage is 120V for about 1.5h.
(2) After step (1) is completed, transferring the protein to a nitrocellulose membrane; the voltage was 16V and the transfer was about 1.2h.
(3) After the completion of the step (2), the nitrocellulose membrane was blocked with 5% nonfat dry milk (TBST solution as solvent) at room temperature for 1h.
TBST solution: tris 2.42g, naCl 8g and Tween-20 ml were dissolved in 1L water and the pH was adjusted to 7.6.
(4) After the step (3) is completed, adding the primary antibody diluted by 5% skimmed milk powder according to a certain proportion on the nitrocellulose membrane, incubating overnight at 4 ℃, and washing the membrane with TBST solution for 3 times each for 5min.
The primary antibody is a rabbit anti-human PFKL antibody (U.S. Santa Cruz Biotech Co.) or a horseradish peroxidase-conjugated beta-actin antibody (U.S. ProteinTech).
(5) After the step (4) is completed, horseradish peroxidase-conjugated IgG (goat anti-rabbit IgG antibody, U.S. Santa Cruz Biotech) diluted with 5% skimmed milk powder in a certain proportion is added to the nitrocellulose membrane, and the membrane is gently shaken at room temperature for 1h, and washed with TBST solution for 3 times, 5min each time.
(6) After the step (5) is completed, the color is developed for 8min by a chemiluminescence method, and the film is pressed and developed.
The results are shown in FIG. 1B (NC is NC siRNA,1 is siRNA-1,2 is siRNA-2,3 is siRNA-3,4 is siRNA-4, and 5 is siRNA-5). The results show that compared with NC siRNA, the expression level of PFKL proteins of siRNA-1, siRNA-2, siRNA-3, siRNA-4 and siRNA-5 transfected human hepatoma cells HepG2 is obviously reduced, wherein the reduction of the expression level of PFKL proteins of siRNA-1 transfected human hepatoma cells HepG2 is most obvious (namely, the interference is the greatest), and the effect is the best.
Example 2 Effect of oligo-nucleic acid siRNA on proliferation of human liver cancer cells
1. Respectively inoculating siRNA transfected human liver cancer cell HepG2 obtained in step two of example 1 to 96-well plates, inoculating about 3000 cells per well, adding DMEM medium containing 10% new born calf serum (day 0 at this time), 37 deg.C, 5% CO 2 Culturing; OD was measured daily with Cell Counting Kit-8 (Dojindo Co., japan) 450nm Values.
On the abscissa of the days of cultivation, the corresponding OD 450nm Values are on the ordinate and growth curves are plotted.
The growth curve is shown in fig. 2 (p < 0.01, p < 0.05), and the experimental result of the growth curve has statistical significance, and p < 0.05. The results show that compared with NC siRNA, siRNA-1, siRNA-2 and siRNA-3 can inhibit proliferation of human hepatoma cell HepG2, and the inhibition effect of siRNA-1 on proliferation of human hepatoma cell HepG2 is most obvious.
Example 3 siRNA-1 inhibiting growth of human liver cancer cells
The influence of siRNA-1 on the growth capacity of human liver cancer cells is studied by adopting a clone formation experiment. The experiment was repeated three times to average the values, and the procedure for each repetition was as follows:
1. inoculating siRNA-1 transfected human liver cancer cell HepG2 obtained in step two of example 1 to 6-well plate, inoculating about 3000 cells per well, adding DMEM medium containing 10% new born calf serum, 37 ℃ and 5% CO 2 Culturing for 2 weeks.
2. After completion of step 1, the cells were gently washed with PBS buffer, then fixed with 4% paraformaldehyde, and stained with 0.1% crystal violet, and counted for colony formation.
According to the steps, the siRNA-1 transfected human liver cancer cell HepG2 is replaced by NC siRNA transfected human liver cancer cell HepG2, and other steps are unchanged.
The cell status of the colony formation assay is shown in FIG. 3A. The statistics of colony formation counts are shown in fig. 3B (x represents p < 0.01). The results show that compared with NC siRNA, the growth capacity of the siRNA-1 transfected human liver cancer cell HepG2 is obviously reduced, and the quantity of the siRNA-1 transfected human liver cancer cell HepG2 is obviously reduced.
The above results indicate that the clone formation count of siRNA-1 transfected human hepatoma cell HepG2 is significantly reduced (p < 0.01) compared to NC siRNA. The siRNA-1 can inhibit the clone formation of the human liver cancer cell HepG2, namely the siRNA-1 can inhibit the growth of the human liver cancer cell.
Example 4 siRNA-1 inhibiting migration of human liver cancer cells
The influence of siRNA-1 on the migration capacity of human liver cancer cells is studied through a cell scratch experiment. The experiment was repeated three times to average the values, and the procedure for each repetition was as follows:
1. the siRNA-1 transfected human hepatoma cells HepG2 obtained in step two of example 1 were inoculated into 6-well plates, each of which was inoculated with about 1X 10 6 The cells were then added with DMEM medium containing 10% new born calf serum at 37℃with 5% CO 2 Culturing until the cell fusion density reaches 90%.
2. After the step 1 is completed, firstly, the tip of a pipette with the specification of 200 mu L is scratched, then, the pipette is washed twice by PBS buffer solution, the scratched cells are removed, and thenAdding into serum-free DMEM medium, 37 deg.C, 5% CO 2 Culturing. And respectively culturing for 0h and 24h, observing and photographing under a white light inverted microscope, and counting the migration distance to be used as an experimental group.
3. According to the steps 1 and 2, the siRNA-1 transfected human liver cancer cell HepG2 is replaced by NC siRNA transfected human liver cancer cell HepG2, other steps are unchanged, and the migration distance is counted to be used as a control group.
4. Relative cell migration was counted. Relative cell migration = migration distance/control migration distance. I.e., the relative cell migration of the control group was taken as 1, and the relative cell migration of the experimental group was counted.
The cell status of the scratch assay is shown in FIG. 4A. The relative cell migration statistics are shown in fig. 4, B (x represents p < 0.01). The result shows that the cell migration capability of the siRNA-1 transfected human liver cancer cell HepG2 is obviously reduced and the scratch healing capability is obviously reduced compared with NC siRNA after 24 hours of culture after scratch. siRNA-1 can inhibit migration of human liver cancer cells.
Example 5 siRNA-1 decreasing glycolytic Capacity of human liver cancer cells
To investigate whether siRNA-1 affects glycolytic function of human hepatoma cells, lactic acid level and ATP yield of siRNA-1 transfected human hepatoma cells HepG2 obtained in step two of example 1 and NC siRNA transfected human hepatoma cells HepG2 were detected using a lactic acid detection kit (Biovision Co., USA) and an ATP content detection kit (Biovision Co., USA), respectively.
The experimental results are shown in fig. 5 (p < 0.01). The results show that compared with NC siRNA, the lactic acid level and ATP content of the human hepatoma cell HepG2 transfected by siRNA-1 are obviously reduced, namely, the siRNA-1 can reduce the glycolytic capacity of the human hepatoma cell HepG2.
The present application is described in detail above. It will be apparent to those skilled in the art that the present application can be practiced in a wide range of equivalent parameters, concentrations, and conditions without departing from the spirit and scope of the application and without undue experimentation. While the application has been described with respect to specific embodiments, it will be appreciated that the application may be further modified. In general, this application is intended to cover any variations, uses, or adaptations of the application following, in general, the principles of the application and including such departures from the present disclosure as come within known or customary practice within the art to which the application pertains.
Claims (8)
1. Oligonucleotide siRNA-1; the oligonucleotide siRNA-1 consists of a single-stranded nucleic acid molecule 1 and a single-stranded nucleic acid molecule 2; the single-stranded nucleic acid molecule 1 consists of SEQ ID NO:1 and 2 dT, wherein the 2 dT is positioned at the 3' end; the single-stranded nucleic acid molecule 2 consists of SEQ ID NO:2, and a single-stranded RNA molecule shown in seq id no.
2. Use of the oligonucleotide siRNA-1 of claim 1 in the manufacture of a medicament for preventing liver cancer.
3. Use of the oligonucleotide siRNA-1 of claim 1 in the manufacture of a medicament for the treatment of liver cancer.
4. The use of the oligonucleotide siRNA-1 of claim 1 in the manufacture of a medicament for inhibiting proliferation of liver cancer cells.
5. Use of the oligonucleotide siRNA-1 of claim 1 in the manufacture of a medicament for inhibiting growth of a liver cancer cell.
6. The use according to claim 5, wherein: the inhibition of the growth of liver cancer cells is shown as inhibition of the clone formation of liver cancer cells.
7. Use of the oligonucleotide siRNA-1 of claim 1 in the manufacture of a medicament for inhibiting migration of liver cancer cells.
8. Use according to any one of claims 2 to 7, characterized in that: the liver cancer is liver cell liver cancer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310531654.6A CN116286828B (en) | 2023-05-12 | 2023-05-12 | Oligonucleotide siRNA and application thereof in preparation of drugs for preventing and treating liver cancer |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310531654.6A CN116286828B (en) | 2023-05-12 | 2023-05-12 | Oligonucleotide siRNA and application thereof in preparation of drugs for preventing and treating liver cancer |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116286828A CN116286828A (en) | 2023-06-23 |
| CN116286828B true CN116286828B (en) | 2023-08-18 |
Family
ID=86803422
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310531654.6A Active CN116286828B (en) | 2023-05-12 | 2023-05-12 | Oligonucleotide siRNA and application thereof in preparation of drugs for preventing and treating liver cancer |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116286828B (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103189391A (en) * | 2010-09-03 | 2013-07-03 | 葛兰素史密斯克莱知识产权发展有限公司 | Novel antigen binding proteins |
| CN104144707A (en) * | 2011-12-14 | 2014-11-12 | 得克萨斯系统大学董事会 | Collateral gene inactivation biomarkers and targets for cancer therapy |
| CN104531709A (en) * | 2014-12-24 | 2015-04-22 | 葛银林 | siRNA dual-interference composition for inhibiting growth and metastasis of tumors and application thereof |
| CN112639083A (en) * | 2018-08-31 | 2021-04-09 | 诺华股份有限公司 | Method for producing cells expressing chimeric antigen receptor |
| CN113862273A (en) * | 2021-12-06 | 2021-12-31 | 中国人民解放军军事科学院军事医学研究院 | HK2 low-expression breast cancer cell line and siRNA used by same |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2008026946A2 (en) * | 2006-08-30 | 2008-03-06 | Genesis Research And Development Corporation Limited | Compositions and methods for the treatment and prevention of neoplastic disorders |
| WO2014028939A2 (en) * | 2012-08-17 | 2014-02-20 | California Institute Of Technology | Targeting phosphofructokinase and its glycosylation form for cancer |
| AU2018214601A1 (en) * | 2017-02-02 | 2019-08-15 | Caris Science, Inc. | Targeted oligonucleotides |
-
2023
- 2023-05-12 CN CN202310531654.6A patent/CN116286828B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103189391A (en) * | 2010-09-03 | 2013-07-03 | 葛兰素史密斯克莱知识产权发展有限公司 | Novel antigen binding proteins |
| CN104144707A (en) * | 2011-12-14 | 2014-11-12 | 得克萨斯系统大学董事会 | Collateral gene inactivation biomarkers and targets for cancer therapy |
| CN104531709A (en) * | 2014-12-24 | 2015-04-22 | 葛银林 | siRNA dual-interference composition for inhibiting growth and metastasis of tumors and application thereof |
| CN112639083A (en) * | 2018-08-31 | 2021-04-09 | 诺华股份有限公司 | Method for producing cells expressing chimeric antigen receptor |
| CN113862273A (en) * | 2021-12-06 | 2021-12-31 | 中国人民解放军军事科学院军事医学研究院 | HK2 low-expression breast cancer cell line and siRNA used by same |
Non-Patent Citations (1)
| Title |
|---|
| "A20 targets PFKL and glycolysis to inhibit theprogression of hepatocellular carcinoma";Yilu Feng等;《Cell Death & Disease》;第2卷(第11期);1-15 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116286828A (en) | 2023-06-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CA2804599C (en) | Mirna and its diagnostic and therapeutic uses in diseases or conditions associated with melanoma, or in diseases or conditions associated with activated braf pathway | |
| EP3199165A1 (en) | Methods and compositions for the specific inhibition of kras by asymmetric double-stranded rna | |
| US9944932B2 (en) | MiRNA molecule defined by its source and its diagnostic and therapeutic uses in diseases or conditions associated with EMT | |
| CN111118011B (en) | siRNA for inhibiting expression of hsa _ circ _0027478 and application thereof | |
| US10612020B2 (en) | Artificial mimic miRNA for controlling gene expression, and use of same | |
| WO2012006181A2 (en) | Compositions and methods for inhibiting oncogenic micrornas and treatment of cancer | |
| CN111118012B (en) | siRNA for inhibiting expression of hsa _ circ _0051680 and application thereof | |
| CN109929841B (en) | siRNA for inhibiting circ _0006033 expression and application thereof | |
| US20240124880A1 (en) | DIAGNOSTIC KIT FOR METASTASIS AND INVASION OF BREAST CANCER AND USE OF shRNA MOLECULE FOR SILENCING EXPRESSION OF HUMAN LINC01614 | |
| CN106668861A (en) | Method for enhancing anti-cervical cancer activity of glutaminase enzyme inhibitor by taking TIGA1 (Transcript Induced by Growth Arrest 1) as target spot | |
| CN116286828B (en) | Oligonucleotide siRNA and application thereof in preparation of drugs for preventing and treating liver cancer | |
| CN109825504B (en) | siRNA for inhibiting circ _0001016 expression and application thereof | |
| CN109745335B (en) | Application of miR-218 in preparation of breast cancer chemotherapeutic drug sensitizer | |
| CN109706152B (en) | siRNA for inhibiting circ _0001017 expression and application thereof | |
| CN107693535A (en) | A kind of microRNA application | |
| CN113862273B (en) | HK2 low-expression breast cancer cell line and siRNA used by same | |
| CN104147616B (en) | The application of double-stranded RNA or its trim in tumor inhibitor is prepared | |
| CN110628791B (en) | Application of tRNA (transfer RNA) modified enzyme gene in non-small cell lung cancer | |
| US9540644B2 (en) | Small interference RNA for inhibiting intracellular expression of ribosomal protein S3 | |
| CN114452294B (en) | Small nucleic acid medicine for treating hepatic fibrosis | |
| CN101538569B (en) | RNA and recombinant for inhibiting human DMT1 protein and application thereof | |
| CN108192895B (en) | siRNA molecule targeting NOB1 gene and application thereof | |
| CN111647598B (en) | siRNA for inhibiting expression of hsa _ circ _0027477 and application thereof | |
| US20220096516A1 (en) | Oligonucleotide molecule and application thereof in tumor therapy | |
| US10351855B2 (en) | Pharmaceutical composition for inhibition of cancer cell metastasis including siRNA for ribosomal protein S3 |
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 |