CN109999199A - Application of the tiRNA as drug target in colorectal cancer transfer treatment - Google Patents

Application of the tiRNA as drug target in colorectal cancer transfer treatment Download PDF

Info

Publication number
CN109999199A
CN109999199A CN201910215582.8A CN201910215582A CN109999199A CN 109999199 A CN109999199 A CN 109999199A CN 201910215582 A CN201910215582 A CN 201910215582A CN 109999199 A CN109999199 A CN 109999199A
Authority
CN
China
Prior art keywords
tirna
colorectal cancer
seq
rna
val
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.)
Withdrawn
Application number
CN201910215582.8A
Other languages
Chinese (zh)
Inventor
盛静浩
许正平
李斯琪
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Zhejiang University ZJU
Original Assignee
Zhejiang University ZJU
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by Zhejiang University ZJU filed Critical Zhejiang University ZJU
Priority to CN201910215582.8A priority Critical patent/CN109999199A/en
Publication of CN109999199A publication Critical patent/CN109999199A/en
Withdrawn legal-status Critical Current

Links

Classifications

    • 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
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K45/00—Medicinal preparations containing active ingredients not provided for in groups A61K31/00 - A61K41/00
    • 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
    • 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
    • C12Q2600/106—Pharmacogenomics, i.e. genetic variability in individual responses to drugs and drug metabolism
    • 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

  • Health & Medical Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • General Health & Medical Sciences (AREA)
  • Organic Chemistry (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Veterinary Medicine (AREA)
  • Medicinal Chemistry (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Immunology (AREA)
  • Wood Science & Technology (AREA)
  • Biochemistry (AREA)
  • Epidemiology (AREA)
  • Zoology (AREA)
  • Molecular Biology (AREA)
  • Pathology (AREA)
  • Genetics & Genomics (AREA)
  • Analytical Chemistry (AREA)
  • Oncology (AREA)
  • Engineering & Computer Science (AREA)
  • Biophysics (AREA)
  • Physics & Mathematics (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Hospice & Palliative Care (AREA)
  • General Chemical & Material Sciences (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The invention belongs to biomedicine fields, it is related to a new colorectal cancer transfer therapy target, shifts therapy target in particular to using tiRNA as colorectal cancer, tiRNA and its antagonist siRNA is further related to, including using them to prepare the application that drug is shifted as targeted therapy colorectal cancer.The invention discloses the purposes of tiRNA: shifting therapy target as colorectal cancer or as the medicament of regulation, treatment colorectal cancer transfer.The antagonist of 5'-tiRNA great clinical value in preparation treatment colorectal cancer pharmaceutical composition provides new drug and method for effective treatment of colorectal cancer.

Description

Application of the tiRNA as drug target in colorectal cancer transfer treatment
Technical field
The invention belongs to biomedicine fields, are related to a new colorectal cancer transfer therapy target, in particular to Therapy target is shifted using tiRNA as colorectal cancer, further relates to tiRNA and its antagonist siRNA, including them is used to prepare medicine The application that object is shifted as targeted therapy colorectal cancer.
Background technique
Malignant tumour is to seriously endanger the disease of human life and health, annual about 3,070,000 people of pathogenesis of cancer number in China, and Every year because of about 2,200,000 people of the number of cancer mortality.World Health Organization's 2012 " report of world's cancer " points out Chinese new diagnosis Cases of cancer is 3,070,000, accounts for the 21.8% of global sum；And year death toll 2,200,000, account for global cancer year death toll 26.9%.More seriously these data just increase year by year at an amazing speed.Therefore, the molecule machine that cancer occurs is explored System is studied and finds its effective treatment means and is of great significance to human health.
Colorectal cancer (Colorectal Cancer, CRC) is that a kind of alimentary canal for seriously threatening human health is pernicious swollen Tumor, morbidity and mortality rank among the best.U.S. CRC disease incidence position and the death rate occupy malignant tumour third position.China CRC Total incidence occupies malignant tumour the 4th, and general mortality rate occupies the 5th.Colorectal cancer incidence rate is obviously high in developed country In developing country, the factors such as this is with developed country higher obesity rates, unsound eating habit are related.2 months 2018, state The Nattonal Cancer statistical data of family's Cancer center's newest phase shows that east China area colorectal cancer incidence rate, the death rate are most Height, this may be related with the obvious westernization of crowd's life style of east developed regions.Therefore, with national life habit and diet side The change of formula, the morbidity and mortality of colorectal cancer are in rapid increase, and treatment needs to improve.
Metastases are complicated, multi-step a biological processes, are related to the abnormal change of many genes adjusting.Closely Over year the study found that non-coding RNA (ncRNA) plays an important role during metastases, the exception of many ncRNA It is directly related with metastases or recurrence.With the development and application of high throughput sequencing technologies, researcher is in tumor patient The small non-coding RNA segment (tsRNA) in the source multiple types tRNA is had found in tissue sample and serum.It is more noticeable It is that they are to shear generation under given conditions by tRNA, but its abundance is not again directly proportional between the tRNA of source, prompts this The generation of a little small RNA fragments is a process that is orderly, strictly being regulated and controled, and participates in specific biological process.It is mature The secondary structure of tRNA is trifolium-shaped, can be divided into and receive stem wall, D ring, T ψ C ring, anticodon loop and variable loop.According to tRNA The length of product and the characteristic of shearing site are sheared, tsRNA is broadly divided into two major classes: one kind is at the anticodon loop of tRNA The tRNA segment generated by the shearing of specific endoribonuclease, referred to as tiRNA (tRNA-derived stress-induced RNA), wherein the tiRNA at the end 5' is known as 5'-tiRNA, and length is 30-35 nucleotide (nts), and the tiRNA at the end 3' is known as 3'- TiRNA, length are 40-50 nucleotide (nts).Existing research shows the Rny1 albumen in fission yeast and the blood in people's cell It is the endoribonuclease for participating in the shearing of tRNA anticodon loop position that pipe, which generates plain (angiogenin, ANG),.TiRNA master It is positioned in cytoplasm, is present in nucleus and mitochondria on a small quantity, and can also be detected in blood circulation of human body system To its presence.Another kind of is the derivative products of mature tRNA or pre-tRNA that length is 12-30 nucleotide (nts), quilt Referred to as tRF (tRNA derived fragment).Currently, we are to tsRNA, effect of the especially tiRNA in colorectal cancer Mechanism and clinical meaning are still unclear, need to be clarified.
Summary of the invention
The technical problem to be solved in the present invention is to provide a kind of tiRNA as drug target in colorectal cancer transfer medicine Application in object.
In order to solve the above technical problem, the present invention provides the purposes of tiRNA a kind of: shifting therapeutic target as colorectal cancer Point or the medicament shifted as regulation, treatment colorectal cancer.
The improvement of purposes as tiRNA of the invention: tiRNA includes 5'-tiRNA and 3'-tiRNA；
5'-tiRNA is following any: 5'-tiRNA-Val-AAC, 5'-tiRNA-Ala-AGC, 5'-tiRNA-Pro- AGG、5'-tiRNA-Val-CAC、5'-tiRNA-Leu-CAG、5'-tiRNA-Glu-CTC、5'-tiRNA-Cys-GCA、5'- TiRNA-Gly-TCC, 5'-tiRNA-His-GTG, 5'-tiRNA-SeC-TCA and 5'-tiRNA-Tyr-GTA；
3'-tiRNA is following any: 3'-tiRNA-Ala-TGC, 3'-tiRNA-Ala-CGC, 3'-tiRNA-Gly- CCC、3'-tiRNA-Val-TAC、3'-tiRNA-Ala-AGC、3'-tiRNA-Met-CAT、3'-tiRNA-Thr-TGT、3'- TiRNA-Trp-CCA, 3'-tiRNA-Arg-ACG and 3'-tiRNA-Arg-CCT.
The further improvement of purposes as tiRNA of the invention: the antagonist siRNA in medicament containing tiRNA.
The siRNA of 5'-tiRNA-Val-AAC is that the siRNA of SEQ ID NO.22,5'-tiRNA-Ala-AGC are SEQ ID The siRNA of NO.23,5'-tiRNA-Cys-GCA are SEQ ID NO.24.
Application the present invention provides tiRNA as drug target in colorectal cancer transfer therapeutic agent, the tiRNA For the small non-coding RNA in transfer RNA (tRNA) source, it is specific as SEQ ID NO.1, SEQ ID NO.2, SEQ ID NO.3, SEQ ID NO.4、SEQ ID NO.5、SEQ ID NO.6、SEQ ID NO.7、SEQ ID NO.8、SEQ ID NO.9、SEQ ID NO.10、SEQ ID NO.11、SEQ ID NO.12、SEQ ID NO.13、SEQ ID NO.14、SEQ ID NO.15、SEQ ID NO.16, SEQ ID NO.17, SEQ ID NO.18, SEQ ID NO.19, SEQ ID NO.20, shown in SEQ ID NO.21 RNA sequence.
The further improvement of the application of tiRNA of the invention as drug target in colorectal cancer transfer therapeutic agent: The method that the targeting changes tiRNA expression.This method includes that siRNA transfection is mediated by transfection reagent, will be directed to upper The siRNA for stating tiRNA is transfected to colorectal cancer cell system, corresponding tiRNA expression in silencing colorectal cancer cell；Or pass through Transfection reagent mediates artificial chemical synthesis tiRNA transfection, raises corresponding tiRNA expression in colorectal cancer cell.
The feature of sequence of the present invention is as follows:
The sequence signature of SEQ ID NO.1: GUUUCCGUAGUGUAGUGGUCAUCACGUUCGCC
Length: 32 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 5'- tiRNA-Val-AAC；Source: human mature tRNA-Val-AAC
The sequence signature of SEQ ID NO.2: GGUGGUGUAGCUCAGUGGUAGAGCGCGUGC
Length: 30 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 5'- tiRNA-Ala-AGC；Source: human mature tRNA-Ala-AGC
The sequence signature of SEQ ID NO.3: GGCUCGUUGGUCUAGGGGUAUGAUUCUCGCUU
Length: 32 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 5'- tiRNA-Pro-AGG；Source: human mature tRNA-Pro-AGG
The sequence signature of SEQ ID NO.4: GUUUCCGUAGUGUAGUGGUUAUCACGUUCGCC
Length: 32 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 5'- tiRNA-Val-CAC；Source: human mature tRNA-Val-CAC
The sequence signature of SEQ ID NO.5: GUCAGGAUGGCCGAGCGGUCUAAGGCGCUGCGUUC
Length: 35 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 5'- tiRNA-Leu-CAG；Source: human mature tRNA-Leu-CAG
The sequence signature of SEQ ID NO.6: UCCCUGGUGGUCUAGUGGUUAGGAUUCGGCA
Length: 31 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 5'- tiRNA-Glu-CTC；Source: human mature tRNA-Glu-CTC
The sequence signature of SEQ ID NO.7: GGGGUAUAGCUCAGUGGUAGAGCAUUUGA
Length: 29 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 5'- tiRNA-Cys-GCA；Source: human mature tRNA-Cys-GCA
The sequence signature of SEQ ID NO.8: GCGUUGGUGGUAUAGUGGUUAGCAUAGCUGCC
Length: 32 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 5'- tiRNA-Gly-TCC；Source: human mature tRNA-Gly-TCC
The sequence signature of SEQ ID NO.9: GCCGUGAUCGUAUAGUGGUUAGUACUCUGCGUU
Length: 33 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 5'- tiRNA-His-GTG；Source: human mature tRNA-His-GTG
The sequence signature of SEQ ID NO.10: GCCCGGAUGAUCCUCAGUGGUCUGGGGUGCAGGC
Length: 34 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 5'- tiRNA-SeC-TCA；Source: human mature tRNA-SeC-TCA
The sequence signature of SEQ ID NO.11: GCCCGGAUGAUCCUCAGUGGUCUGGGGUGCAGGC
Length: 34 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 5'- tiRNA-Tyr-GTA；Source: human mature tRNA-Tyr-GTA
The sequence signature of SEQ ID NO.12: GUAUGAGGCCCCGGGUUCAAUCCCCGGCAUCUCCACCA
Length: 38 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 3'- tiRNA-Ala-TGC；Source: human mature tRNA-Ala-TGC
The sequence signature of SEQ ID NO.13: GUAUGAGGCCCCGGGUUCGAUCCCCGGCAUCUCCACCA
Length: 38 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 3'- tiRNA-Ala-CGC；Source: human mature tRNA-Ala-CGC
The sequence signature of SEQ ID NO.14: AUUCUUGCGACCCGGGUUCGUUUCCCGGGCGGCGCACCA
Length: 39 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 3'- tiRNA-Gly-CCC；Source: human mature tRNA-Gly-CCC
The sequence signature of SEQ ID NO.15: ACACGCAGAAGGUCCTGGGUUCGAGCCCCAGUGGAACCACCA
Length: 42 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 3'- tiRNA-Val-TAC；Source: human mature tRNA-Val-TAC
The sequence signature of SEQ ID NO.16: GCAUGAGGUCCCGGGUUCGAUCCCCAGCAUCUCCACCA
Length: 38 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 3'- tiRNA-Ala-AGC；Source: human mature tRNA-Ala-AGC
The sequence signature of SEQ ID NO.17: UCUGAAGGUCGUGAGUUCGAGCCUCACACGGGGCACU
Length: 37 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 3'- tiRNA-Met-CAT；Source: human mature tRNA-Met-CAT
The sequence signature of SEQ ID NO.18: CCAGGGGUCGCGAGUUCAUUUCUCGCUGGGGCCUCCA
Length: 37 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 3'- tiRNA-Thr-TGT；Source: human mature tRNA-Thr-TGT
The sequence signature of SEQ ID NO.19: UCAGAAGGCUGCGUGUUCGAAUCACGUCGGGGUCACC
Length: 37 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 3'- tiRNA-Trp-CCA；Source: human mature tRNA-Trp-CCA
The sequence signature of SEQ ID NO.20: UCAGAAGAUUCUAGGUUCGACUCCUGGCUGGCUCGCA
Length: 37 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 3'- tiRNA-Arg-ACG；Source: human mature tRNA-Arg-ACG
The sequence signature of SEQ ID NO.21: CCAGGGAUUGUGGGUUCGAGUCCCAUCUGGGGUGCU
Length: 37 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: 3'- tiRNA-Arg-CCT；Source: human mature tRNA-Arg-CCT
The sequence signature of SEQ ID NO.22: GUAGUGUAGUGGUUAUCACU
Length: 20 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: siRNA or shRNA；Source: 5'-tiRNA-Val-AAC
The sequence signature of SEQ ID NO.23: GUAGCUCAGUGGUAGAGCUU
Length: 20 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: siRNA or shRNA；Source: 5'-tiRNA-Ala-AGC
The sequence signature of SEQ ID NO.24: AUAGCUCAGUGGUAGAGCUU
Length: 20 nucleotide；Type: ribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: siRNA or shRNA；Source: 5'-tiRNA-Cys-GCA
The sequence signature of SEQ ID NO.25: AGGCGAACGTGATAACCACTACACTACGGA
Length: 30 nucleotide；Type: deoxyribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: DIG Probe；Source: 5'-tiRNA-Val-AGC
The sequence signature of SEQ ID NO.26: GCACGCGCTCTACCACTGAGCTACACCCCC
Length: 30 nucleotide；Type: deoxyribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: DIG Probe；Source: 5'-tiRNA-Ala-AGC
The sequence signature of SEQ ID NO.27: TCAAATGCTCTACCACTGAGCTATACCCCC
Length: 20 nucleotide；Type: deoxyribonucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: DIG Probe；Source: 5'-tiRNA-Cys-GCA
The sequence signature of SEQ ID NO.28: UGUGAGUCACGUGAGGGCAGAAUCUGCUC
Length: 29 nucleotide；Type: nucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: tiRNA；Source: The small non-coding RNA -1, tiRNA-ctrl of random controls
The sequence signature of SEQ ID NO.29: UUCUCCGAACGUGUCACGU
Length: 19 nucleotide；Type: nucleotide；Chain: single-stranded；Topological structure: line style；Characteristic: siRNA or shRNA；Source: the small non-coding RNA -2, siRNA-ctrl of random controls
The solution of the present invention is specific as follows:
The present invention provides a new colorectal cancers to shift therapy target --- tiRNA, including 5'-tiRNA and 3'- tiRNA.Specifically, using high throughput sequencing technologies analysis compare it is poor in 3 colorectal cancer patients cancer beside organisms and cancerous tissue The small non-coding RNA (as shown in Figure 1) of different expression, wherein the highly expressed 5'-tiRNA of conspicuousness includes in Colorectal Carcinoma 5'-tiRNA-Val-AAC (nucleic acid sequence is as shown in SEQ ID NO.1), 5'-tiRNA-Ala-AGC (nucleic acid sequence such as SEQ ID Shown in NO.2), 5'-tiRNA-Pro-AGG (nucleic acid sequence is as shown in SEQ ID NO.3), 5'-tiRNA-Val-CAC (nucleic acid sequence Column as shown in SEQ ID NO.4), 5'-tiRNA-Leu-CAG (nucleic acid sequence is as shown in SEQ ID NO.5), 5'-tiRNA- Glu-CTC (nucleic acid sequence is as shown in SEQ ID NO.6) and 5'-tiRNA-Cys-GCA (nucleic acid sequence such as SEQ ID NO.7 institute Show)；The 5'-tiRNA of conspicuousness low expression includes 5'-tiRNA-Gly-TCC (nucleic acid sequence is as shown in SEQ ID NO.8), 5'- TiRNA-His-GTG (nucleic acid sequence is as shown in SEQ ID NO.9), 5'-tiRNA-SeC-TCA (nucleic acid sequence such as SEQ ID Shown in NO.10), 5'-tiRNA-Tyr-GTA (nucleic acid sequence is as shown in SEQ ID NO.11)；The highly expressed 3'- of conspicuousness TiRNA includes 3'-tiRNA-Ala-TGC (nucleic acid sequence is as shown in SEQ ID NO.12), 3'-tiRNA-Ala-CGC (nucleic acid sequence Column as shown in SEQ ID NO.13), 3'-tiRNA-Gly-CCC (nucleic acid sequence is as shown in SEQ ID NO.14), 3'-tiRNA- Val-TAC (nucleic acid sequence is as shown in SEQ ID NO.15), 3'-tiRNA-Ala-AGC (nucleic acid sequence such as SEQ ID NO.16 institute Show), 3'-tiRNA-Met-CAT (nucleic acid sequence is as shown in SEQ ID NO.17)；The 3'-tiRNA of conspicuousness low expression includes 3'-tiRNA-Thr-TGT (nucleic acid sequence is as shown in SEQ ID NO.18), 3'-tiRNA-Trp-CCA (nucleic acid sequence such as SEQ Shown in ID NO.19), 3'-tiRNA-Arg-ACG (nucleic acid sequence is as shown in SEQ ID NO.20), 3'-tiRNA-Arg-CCT (nucleic acid sequence is as shown in SEQ ID NO.21).
Further, specific 5'-tiRNA is demonstrated using RNA immunoblot method to be significantly higher than in Expression in Colorectal Cancer Cancer beside organism (shows) as described in Figure 2, specifically includes 5'-tiRNA-Val-AAC, 5'-tiRNA-Ala-AGC and 5'-tiRNA- Cys-GCA.Meanwhile comparing high transfer and low transfer colorectal cancer cell lines (HCT116) difference from same genetic background 5'-tiRNA expression (such as Fig. 3 is shown), as the result is shown expression water of the above-mentioned three kinds of 5'-tiRNA in high-transfer cell strain It is flat to be significantly higher than low metastatic cells.
The present invention provides a kind of methods for changing intracellular tiRNA expression, including artificial chemistry synthesis tiRNA to mention Its high expression or siRNA inhibit tiRNA expression.Wherein, term " siRNA " refers to short interfering rna (Short Interfering RNA, referred to as " siRNA "), it is the double-stranded RNA of 20 to 25 nucleotide of length, the mode for capableing of specificity presses down The expression of gene processed.According to the type of tiRNA, different siRNA is devised, wherein 5'-tiRNA-Val-AAC, 5'-tiRNA- The siRNA/shRNA acting sequences of Ala-AGC and 5'-tiRNA-Cys-GCA such as SEQ ID NO.22, SEQ ID NO.23 and Shown in SEQ ID NO.24.However, artificial chemistry synthesis tiRNA and its siRNA involved in the present invention is not limited to above-mentioned three Sequence further includes the tiRNA positioned at above-mentioned other differential expressions.
The method of targeted inhibition colorectal cancer transfer provided by the invention, this method are the level of targeted inhibition tiRNA.
The beneficial effects of the present invention are:
Compared with prior art, the present invention is controlled using a new class of small non-coding RNA (tiRNA) as colorectal cancer transfer The target spot for the treatment of, so that all colorectal cancer patients clinical Benefits.The siRNA of tiRNA is used to prepare inhibition knot directly by the present invention Intestinal cancer transfer drug, overcome to colorectal cancer transfer it is non-selective, reach targeted therapy colorectal cancer, improve patient it is pre- Purpose afterwards compensates for the deficiency of the local treatments means such as operation, chemicotherapy, can play a role to micrometastasis stove；Can with gram Chemotherapy is taken to the toxic side effect of body, expands the range of targeted therapy.
The current prior art, 5'-tiRNA and 3'-tiRNA are to the function in tumour (including non-colorectal cancer tumour) transfer It can be unknown, it is unclear that.
In conclusion the present invention discloses a new colorectal cancer transfer therapeutic agent target spot.Pass through a series of internal, bodies The discovery of outer functional experiment: specific 5'-tiRNA (including 5'-tiRNA-Val-AAC, 5'-tiRNA-Ala-AGC and 5'-tiRNA- Cys-GCA migration and the invasive procedure of colorectal cancer cell) can be promoted, and inhibit these 5'-tiRNA that Colon and rectum can be effectively suppressed The migration of cancer cell and invasive procedure.These experimental datas suffice to show that the intracellular 5'-tiRNA level of modulate tumor can be significant Cell migration and invasive ability are reduced, there is the potential as oncotherapy Effective target site.It is prepared by the antagonist of 5'-tiRNA Treat great clinical value in colorectal cancer pharmaceutical composition, for colorectal cancer effective treatment provide new drug with Method.
Detailed description of the invention
Specific embodiments of the present invention will be described in further detail with reference to the accompanying drawing.
Fig. 1 is colorectal cancer patients cancer beside organism and cancerous tissue differential expression tiRNA analysis chart；
A. in colorectal cancer patients cancer beside organism and cancerous tissue small non-coding RNA distribution situation, wherein abscissa is small Non-coding RNA length (coverage area is 16 nucleotide to 49 nucleotide), ordinate is the small non-coding RNA of each length Number is read in sequencing；Light histogram is shown as cancer beside organism, and dark histogram is shown as cancerous tissue；
B. in colorectal cancer patients cancer beside organism and cancerous tissue differential expression tiRNA thermal map.
Fig. 2 is the tiRNA figure of differential expression in RNA trace detection colorectal cancer patients cancer beside organism and cancerous tissue；
5'-tiRNA-Val-AAC, 5'- in 3 colorectal cancer patients cancer beside organisms of rna blot analysis and cancerous tissue The expression of tiRNA-Ala-AGC and 5'-tiRNA-Cys-GCA.
Fig. 3 is the colorectal cancer cell lines figure of the high and low migration invasive ability of screening；
A. high and low migration invasion cell screens schematic diagram, wherein height migration invasion cell is labeled as HCT116-H, low to move It moves invasion cell and is labeled as HCT116-L；
B. migration and the invasive ability of above two subcellular system are detected using Transwell method, picture is representativeness The microscope photo of cell migration and invasion；
C. two kinds of subcellular system migrations and invasion cell statistical result, ordinate show to be thin under each field of microscope The average value of born of the same parents' quantity；
5'-tiRNA-Val-AAC, 5'-tiRNA-Ala-AGC and 5'- in two kinds of subcellular systems of D.RNA engram analysis The expression of tiRNA-Cys-GCA.
Fig. 4 is to transfect artificial chemically synthesized tiRNA to promote transfer and the invasive ability figure of colorectal cancer cell；
A. the artificial chemically synthesized tiRNA of low migration invasion cell HCT116-L transfection (including tiRNA-ctrl, 5'- TiRNA-Val-AAC, 5'-tiRNA-Ala-AGC and 5'-tiRNA-Cys-GCA), it is moved using Transwell method detection cell It moves and invasive ability, picture is the microscope photo that each group representativeness cell is migrated and invaded；
B. every group of cell migration and invasion statistical result；
C. the influence that above-mentioned tiRNA grows cell is detected using CCK-8 method.
Fig. 5 is transfer and the invasive ability figure that specific tiRNA antagonist siRNA can inhibit colorectal cancer cell；
A. high migration invasion cell HCT116-H transfection tiRNA antagonist siRNA (including siRNA-ctrl, 5'-tiRNA- Val-AAC, 5'-tiRNA-Ala-AGC and 5'-tiRNA-Cys-GCA), drop efficiency is struck using the detection of RNA trace, wherein Control represents the small non-coding RNA -2, siRNA of random controls and represents specific siRNA；
B. for above-mentioned cell using the detection cell migration of Transwell method and invasive ability, picture is that each group is representative thin The microscope photo of born of the same parents migration and invasion；
C. above-mentioned every group of cell migration and invasion statistical result；
D. the influence for inhibiting above-mentioned tiRNA to grow cell using the detection of CCK-8 method；
Fig. 6 is that 5'-tiRNA-Val-AAC does not influence colorectal cancer cell subcutaneous transplantation tumor growth figure；
A. nude mice by subcutaneous inoculation has transfected the small non-coding RNA -2 (tiRNA-ctrl) of control or 5'-tiRNA-Val-AAC knot Rectum cancer cell, transplantable tumor is taken out in dissection after 4 weeks, and picture is the transplantable tumor photo taken out；
B. two groups of transplanting tumor weight statistical results, there was no significant difference；
C. it is using immunohistochemical analysis transplantable tumor proliferative conditions (Ki67 dyeing) and angiogenesis situation (CD31), picture Representative immunohistochemistry photo；
D.Ki67 positive cell and microvessel density statistical result.
Fig. 7 is that 5'-tiRNA-Val-AAC promotes colorectal cancer cell mouse lung transfer figure；
A. the small non-coding RNA of control (tiRNA-ctrl) or 5'-tiRNA- have been transfected by tail vein injection The colorectal cancer cell of Val-AAC luciferase label, the real-time living imaging of toy show the colonisation of transplantable tumor；
B. luciferase luminous intensity statistical result；
C. lung transplantation tumor grows photo after mouse dissection；
D. hematoxylin eosin staining shows tumour in the positioning of lung.
Specific embodiment
In order to make it easy to understand, specific drawings and examples the present invention is further explained content will be passed through below.Need spy Not, it is noted that specific example and attached drawing are merely to explanation, those skilled in the art can according to illustrating herein, Various modifications and variations are made to the present invention in the scope of the present invention, model of the invention is also included in these modifications and variations In enclosing.
Embodiment 1: tiRNA expression in colorectal cancer patients cancerous tissue
1. colorectal cancer patients cancer beside organism and cancerous tissue differential expression tiRNA analysis
Implementation method: 3 colorectal cancer patients cancer beside organisms and cancer are detected using small non-coding RNA high-flux sequence method TiRNA is expressed in tissue, then the tiRNA of bioinformatic analysis differential expression.
Interpretation of result: as shown in figure 1, cancer beside organism, the highly expressed 5'- of conspicuousness in Colorectal Carcinoma are compared TiRNA include 5'-tiRNA-Val-AAC, 5'-tiRNA-Ala-AGC, 5'-tiRNA-Pro-AGG, 5'-tiRNA-Val-CAC, 5'-tiRNA-Leu-CAG, 5'-tiRNA-Glu-CTC and 5'-tiRNA-Cys-GCA；The 5'-tiRNA packet of conspicuousness low expression Include 5'-tiRNA-Gly-TCC, 5'-tiRNA-His-GTG, 5'-tiRNA-SeC-TCA and 5'-tiRNA-Tyr-GTA；Conspicuousness Highly expressed 3'-tiRNA includes 3'-tiRNA-Ala-TGC, 3'-tiRNA-Ala-CGC, 3'-tiRNA-Gly-CCC, 3'- TiRNA-Val-TAC, 3'-tiRNA-Ala-AGC and 3'-tiRNA-Met-CAT；The 3'-tiRNA of conspicuousness low expression includes 3'-tiRNA-Thr-TGT, 3'-tiRNA-Trp-CCA, 3'-tiRNA-Arg-ACG and 3'-tiRNA-Arg-CCT.Result above Illustrate that the tiRNA of these differential expressions takes part in the occurrence and development process of colorectal cancer.
2.RNA trace detects the tiRNA of differential expression in colorectal cancer patients cancer beside organism and cancerous tissue
Implementation method: using RNA immunoblot method (Northern Blot) method detection colorectal cancer patients cancer beside organism and The tiRNA of differential expression in cancerous tissue mainly includes 5'-tiRNA-Val-AAC, 5'-tiRNA-Ala-AGC and 5'-tiRNA- Cys-GCA.This method mainly utilizes polyacrylamide gel electrophoresis chorista RNA, is then transferred on nylon membrane, then With specific probe, (probe sequence of 5'-tiRNA-Val-AAC, 5'-tiRNA-Ala-AGC and 5'-tiRNA-Cys-GCA are such as Shown in SEQ ID NO.25, SEQ ID NO.26 and SEQ ID NO.27) it is checked.
Reagent involved by RNA immunoblot method and experiment flow are as follows: 1) using 15% urea-denatured polyacrylamide gel (being purchased from U.S. ThermoFisher company, article No. EC68852BOX) chorista RNA；2) after RNA end of the sample, with 150 Lie prostrate/120 minutes progress polyacrylamide gel electrophoresises；3) after electrophoresis, with SYBR Gold Nucleic Acid Gel Stain dyestuff (being purchased from U.S. ThermoFisher company, article No. S11494) dyes polyacrylamide gel electrophoresis glue 15 minutes, It is taken pictures using ultraviolet imager；4) semidry method transferring film: in advance by filter paper, nylon membrane in 0.5 × TBE (5.4 grams of Tris CAS#77- 86-1,2.75 grams of boric acid CAS#10043-35-3,2 milliliters of 0.5M EDTA CAS#60-00-4pH 8.0 are settled to 1 with water Rise) middle immersion 10 minutes, it is successively stacked from as low as top according to the sequence of filter paper, nylon membrane, PAGE glue, filter paper, 300 milli of constant current Peace replaces 25 volts of constant pressure, transferring film 1 hour when voltage rises to 25 volts；5) after transferring film, 0.5 × TBE of nylon membrane is taken out Rinsing, is placed on filter paper and blots, then the one side for having RNA is placed on upward on new filter paper, 120000 μ J/cm2UV crosslinking 1 Secondary, 180 ° of rotation is crosslinked 1 time again, and filter paper, which is wrapped, sets 80 DEG C of roasting films 1 hour；6) after taking out film, using DIG Northern Starter Kit (being purchased from U.S. Roche company, article No. 12039672910) and specificity tiRNA probe in detecting, and use G- The detection of BOX chemiluminescence detector, then carries out data analysis with corresponding software.
Interpretation of result: as shown in figure 2,5'-tiRNA-Val-AAC, 5'- in 3 colorectal cancer patients cancerous tissues The expression quantity of tiRNA-Ala-AGC and 5'-tiRNA-Cys-GCA is above cancer beside organism.Therefore, with containing specificity TiRNA probe can detecte tiRNA expression in clinical Colorectal Carcinoma sample, can be used as the index of auxiliary diagnosis.
Embodiment 2:tiRNA can promote transfer and the invasive ability of colorectal cancer cell
1. screening the colorectal cancer cell lines of high and low migration invasive ability
Implementation method: Matrigel is migrated based on Transwell, the knot for being separately cultured the cell Transwell upper and lower level is straight Colon-cancer cell (HCT116), isolated culture method reference both have been reported that MiR-218inhibits invasion and metastasis of gastric cancer by targeting the Robo1receptor(PLoS Genet 2010； 6:e1000879)；By 3 wheel screenings (screening conditions are whether cell can migrate to the lower layer of the cell Transwell), choose Meeting respectively can migrate to the cell Transwell lower layer and rests on the cell of the cell Transwell upper layer condition, to screen The different subcellular system of two kinds of migration invasive abilities is established, the high HCT116-H for migrating invasive ability and low migration are respectively designated as The HCT116-L Cell subline of invasive ability.Then, 5'-tiRNA-Val-AAC, 5'-tiRNA- are detected using RNA immunoblot method The expression of Ala-AGC and 5'-tiRNA-Cys-GCA.
Reaction reagent involved in RNA immunoblot method and experiment flow are the same as embodiment 1.
Interpretation of result: as shown in figure 3, the migration of the HCT116-H Cell subline of high invasive ability and invasive ability are above The HCT116-L Cell subline of low invasive ability.Further, RNA polyacrylamide gel electrophoresis HCT116-H Cell subline as the result is shown In tiRNA content conspicuousness be higher than HCT116-L Cell subline；Meanwhile RNA immunoblot method also demonstrates 5'-tiRNA-Val- The height of the content of AAC, 5'-tiRNA-Ala-AGC and 5'-tiRNA-Cys-GCA in HCT116-H Cell subline also conspicuousness In HCT116-L Cell subline.Result above prompts tiRNA to take part in migration and the invasive procedure of colorectal cancer cell.
2. transfer and the invasive ability of colorectal cancer cell can be promoted by transfecting artificial synthesized tiRNA
Implementation method: specific tiRNA and small -1 (tiRNA- of non-coding RNA of random controls are synthesized by artificial chemistry Ctrl, nucleic acid sequence is as shown in SEQ ID NO.28), wherein 5'-tiRNA-Val-AAC, 5'-tiRNA-Ala-AGC and 5'- TiRNA-Cys-GCA sequence is as shown in SEQ ID NO.1, SEQ ID NO.2 and SEQ ID NO.7.Above-mentioned artificial chemistry is closed At tiRNA be transfected into the HCT116-L Cell subline of low invasive ability, observe the migration, invasion and proliferative conditions of cell.
Interpretation of result: as shown in figure 4, compared with small -1 group of non-coding RNA of random controls, 5'-tiRNA-Val- has been transfected The migration of AAC, 5'-tiRNA-Ala-AGC and 5'-tiRNA-Cys-GCA cell and invasive ability conspicuousness increase, and cell increases The level of growing is not affected by apparent influence.This illustrates that above-mentioned tiRNA is with the transfer of promotion colorectal cancer cell and to invade small non- Coding RNA.
3. transfer and invasive ability that specificity tiRNA antagonist siRNA can inhibit colorectal cancer cell
Implementation method: siRNA technology silencing 5'-tiRNA- is used in the HCT116-H subcellular system of high invasive ability The expression of Val-AAC, 5'-tiRNA-Ala-AGC and 5'-tiRNA-Cys-GCA, the siRNA of 5'-tiRNA-Val-AAC SiRNA for SEQ ID NO.22,5'-tiRNA-Ala-AGC is the siRNA of SEQ ID NO.23,5'-tiRNA-Cys-GCA For SEQ ID NO.24.Simultaneously include the small non-coding RNA -2 (nucleic acid sequence is as shown in SEQ ID NO.29) of random controls, utilizes RNA trace detects siRNA jamming effectiveness, observes the migration, invasion and proliferative conditions of cell.
Interpretation of result: as shown in figure 5, compared with the control group, having transfected in the cell of siRNA, the content of corresponding tiRNA It is significantly suppressed, but the expression of its maternal maturation tRNA is not affected.As intracellular 5'-tiRNA-Val- After the expression of AAC, 5'-tiRNA-Ala-AGC and 5'-tiRNA-Cys-GCA are suppressed, the HCT116-H's of high invasive ability Cell migration and invasion conspicuousness are suppressed, and cell Proliferation is not affected by influence.This illustrates that the antagonist siRNA of tiRNA can As colorectal cancer metastasis suppressor target spot.
Embodiment 3:5'-tiRNA-Val-AAC can promote the transfer of colorectal cancer cell mouse lung
1.5'-tiRNA-Val-AAC does not influence the growth of colorectal cancer cell subcutaneous transplantation tumor
Implementation method: being tested using Xenografts in nude mice, and table 5'-tiRNA-Val-AAC or random controls is small non- The colorectal cancer cell of coding RNA -1 is inoculated into the left survey dorsal sc of 4 week old or so nude mice, observes subcutaneous transplantation after 1 month Tumor formational situation.
Interpretation of result: as shown in fig. 6, compared with small -1 cellular control unit of non-coding RNA of random controls, height expression 5'- TiRNA-Val-AAC does not dramatically increase growth and its proliferation marker Ki67 and marker of angiogenesis object CD31 of tumour.
2.5'-tiRNA-Val-AAC promotes the transfer of colorectal cancer cell mouse lung
Implementation method: the luciferase of the high expression 5'-tiRNA-Val-AAC of building or the small non-coding RNA -1 of random controls Colorectal cancer cell system (HCT116-Luc) after tail vein injection, raises 8 weeks, utilizes small animal living body imaging system observation knot Carcinoma of the rectum mouse lung transfer case.
Interpretation of result: as shown in fig. 7, the mouse fluorescence intensity for having injected overexpression 5'-tiRNA-Val-AAC cell is significant Higher than the mouse for having injected small -1 cell of non-coding RNA of random controls.Further, dissection mouse analysis lung tumors shift feelings Condition, 5'-tiRNA-Val-AAC can conspicuousness promotion colorectal cancer cell mouse lung transfer as the result is shown.
The above list is only a few specific embodiments of the present invention for finally, it should also be noted that.Obviously, this hair Bright to be not limited to above embodiments, acceptable there are many deformations.Those skilled in the art can be from present disclosure All deformations for directly exporting or associating, are considered as protection scope of the present invention.
Sequence table
<110>Zhejiang University
<120>application of the tiRNA as drug target in colorectal cancer transfer treatment
<160> 29
<170> SIPOSequenceListing 1.0
<210> 1
<211> 32
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 1
guuuccguag uguagugguc aucacguucg cc 32
<210> 2
<211> 30
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 2
ggugguguag cucaguggua gagcgcgugc 30
<210> 3
<211> 32
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 3
ggcucguugg ucuaggggua ugauucucgc uu 32
<210> 4
<211> 32
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 4
guuuccguag uguagugguu aucacguucg cc 32
<210> 5
<211> 35
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 5
gucaggaugg ccgagcgguc uaaggcgcug cguuc 35
<210> 6
<211> 31
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 6
ucccuggugg ucuagugguu aggauucggc a 31
<210> 7
<211> 29
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 7
gggguauagc ucagugguag agcauuuga 29
<210> 8
<211> 32
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 8
gcguuggugg uauagugguu agcauagcug cc 32
<210> 9
<211> 33
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 9
gccgugaucg uauagugguu aguacucugc guu 33
<210> 10
<211> 34
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 10
gcccggauga uccucagugg ucuggggugc aggc 34
<210> 11
<211> 34
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 11
gcccggauga uccucagugg ucuggggugc aggc 34
<210> 12
<211> 38
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 12
guaugaggcc ccggguucaa uccccggcau cuccacca 38
<210> 13
<211> 38
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 13
guaugaggcc ccggguucga uccccggcau cuccacca 38
<210> 14
<211> 39
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 14
auucuugcga cccggguucg uuucccgggc ggcgcacca 39
<210> 15
<211> 42
<212> DNA/RNA
<213>artificial sequence (Artificial Sequence)
<400> 15
acacgcagaa ggucctgggu ucgagcccca guggaaccac ca 42
<210> 16
<211> 38
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 16
gcaugagguc ccggguucga uccccagcau cuccacca 38
<210> 17
<211> 37
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 17
ucugaagguc gugaguucga gccucacacg gggcacu 37
<210> 18
<211> 37
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 18
ccaggggucg cgaguucauu ucucgcuggg gccucca 37
<210> 19
<211> 37
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 19
ucagaaggcu gcguguucga aucacgucgg ggucacc 37
<210> 20
<211> 37
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 20
ucagaagauu cuagguucga cuccuggcug gcucgca 37
<210> 21
<211> 36
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 21
ccagggauug uggguucgag ucccaucugg ggugcu 36
<210> 22
<211> 20
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 22
guaguguagu gguuaucacu 20
<210> 23
<211> 20
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 23
guagcucagu gguagagcuu 20
<210> 24
<211> 20
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 24
auagcucagu gguagagcuu 20
<210> 25
<211> 30
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 25
aggcgaacgt gataaccact acactacgga 30
<210> 26
<211> 30
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 26
gcacgcgctc taccactgag ctacaccccc 30
<210> 27
<211> 30
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 27
tcaaatgctc taccactgag ctataccccc 30
<210> 28
<211> 29
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 28
ugugagucac gugagggcag aaucugcuc 29
<210> 29
<211> 19
<212> RNA
<213>artificial sequence (Artificial Sequence)
<400> 29
uucuccgaac gugucacgu 19

Claims (5)

  1. The purposes of 1.tiRNA, it is characterized in that: shifting therapy target as colorectal cancer or turning as regulation, treatment colorectal cancer The medicament of shifting.
  2. 2. the purposes of tiRNA according to claim 1, it is characterized in that: tiRNA includes 5'-tiRNA and 3'-tiRNA.
  3. 3. the purposes of tiRNA according to claim 2, it is characterized in that 5'-tiRNA is following any:
    5'-tiRNA-Val-AAC、5'-tiRNA-Ala-AGC、5'-tiRNA-Pro-AGG、5'-tiRNA-Val-CAC、
    5'-tiRNA-Leu-CAG、5'-tiRNA-Glu-CTC、5'-tiRNA-Cys-GCA、5'-tiRNA-Gly-TCC、
    5'-tiRNA-His-GTG, 5'-tiRNA-SeC-TCA and 5'-tiRNA-Tyr-GTA.
  4. 4. the purposes of tiRNA according to claim 2, it is characterized in that 3'-tiRNA is following any:
    3'-tiRNA-Ala-TGC、3'-tiRNA-Ala-CGC、3'-tiRNA-Gly-CCC、3'-tiRNA-Val-TAC、
    3'-tiRNA-Ala-AGC、3'-tiRNA-Met-CAT、3'-tiRNA-Thr-TGT、3'-tiRNA-Trp-CCA、
    3'-tiRNA-Arg-ACG and 3'-tiRNA-Arg-CCT.
  5. 5. the purposes of tiRNA according to claims 1 to 4, it is characterized in that: containing the antagonist of tiRNA in medicament siRNA。
CN201910215582.8A 2019-03-21 2019-03-21 Application of the tiRNA as drug target in colorectal cancer transfer treatment Withdrawn CN109999199A (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201910215582.8A CN109999199A (en) 2019-03-21 2019-03-21 Application of the tiRNA as drug target in colorectal cancer transfer treatment

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201910215582.8A CN109999199A (en) 2019-03-21 2019-03-21 Application of the tiRNA as drug target in colorectal cancer transfer treatment

Publications (1)

Publication Number Publication Date
CN109999199A true CN109999199A (en) 2019-07-12

Family

ID=67167593

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201910215582.8A Withdrawn CN109999199A (en) 2019-03-21 2019-03-21 Application of the tiRNA as drug target in colorectal cancer transfer treatment

Country Status (1)

Country Link
CN (1) CN109999199A (en)

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111671907A (en) * 2020-06-12 2020-09-18 广西医科大学第一附属医院 TiRNA-Val antisense strand inhibitor and application thereof
CN111826444A (en) * 2020-07-17 2020-10-27 南京大学 Serum/plasma tsRNA marker related to pancreatic cancer, probe and application thereof
CN113308530A (en) * 2021-05-24 2021-08-27 嘉兴市第一医院 Blood tsRNA marker for herpes zoster, preparation and application
CN113817821A (en) * 2020-06-18 2021-12-21 上海中医药大学附属龙华医院 Colorectal tumorous lesion blood tRF marker and application thereof
CN113862269A (en) * 2021-10-25 2021-12-31 中南大学湘雅三医院 tsRNA molecules and uses thereof

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN111671907A (en) * 2020-06-12 2020-09-18 广西医科大学第一附属医院 TiRNA-Val antisense strand inhibitor and application thereof
CN113817821A (en) * 2020-06-18 2021-12-21 上海中医药大学附属龙华医院 Colorectal tumorous lesion blood tRF marker and application thereof
CN111826444A (en) * 2020-07-17 2020-10-27 南京大学 Serum/plasma tsRNA marker related to pancreatic cancer, probe and application thereof
CN113308530A (en) * 2021-05-24 2021-08-27 嘉兴市第一医院 Blood tsRNA marker for herpes zoster, preparation and application
CN113862269A (en) * 2021-10-25 2021-12-31 中南大学湘雅三医院 tsRNA molecules and uses thereof
CN113862269B (en) * 2021-10-25 2023-12-22 中南大学湘雅三医院 tsRNA molecules and uses thereof

Similar Documents

Publication Publication Date Title
CN109999199A (en) Application of the tiRNA as drug target in colorectal cancer transfer treatment
Wong et al. The microRNA miR-139 suppresses metastasis and progression of hepatocellular carcinoma by down-regulating Rho-kinase 2
Wang et al. FoxM1 expression is significantly associated with cisplatin-based chemotherapy resistance and poor prognosis in advanced non-small cell lung cancer patients
Liang et al. Downregulation of microRNA-206 promotes invasion and angiogenesis of triple negative breast cancer
Nomoto et al. Fusobacterium nucleatum promotes esophageal squamous cell carcinoma progression via the NOD1/RIPK2/NF-κB pathway
Du et al. Sex determining region Y-box 12 (SOX12) promotes gastric cancer metastasis by upregulating MMP7 and IGF1
CN101478978B (en) Delta-tocotrienol treatment and prevention of pancreatic cancer
Ito et al. Polo‐like kinase 1 regulates cell proliferation and is targeted by miR‐593* in esophageal cancer
CN112522394B (en) Novel exosome release related target and application thereof in tumor monitoring and inhibition
CN111154869B (en) Biomarker for liver cancer diagnosis and kit thereof
CN108531596B (en) Application of lncRNA as biomarker in diagnosis and treatment of gastric cancer
CN105435228A (en) Arsenic trioxide antineoplastic new use and anti-tumor preparation
CN104975023B (en) Human cervical carcinoma shifts relevant new long-chain non-coding RNA sequence, separation method and application thereof
US20220125877A1 (en) Method for treating colorectal cancer
Hao et al. lncRNA-CASC7 inhibits the proliferation and migration of colon cancer by negatively regulating the PI3K/Akt signaling pathway
CN107828872A (en) The detection reagent of Pygo2 gene expressions in Wnt signal paths based on peptide nucleic acid probe
CN113564252B (en) New use of methylase METTL3
CN110452989A (en) Application of the biomarker in gastric cancer is detected, diagnosed
CN108998532A (en) A kind of diagnosis and treatment marker of rectal adenocarcinoma
Chen et al. miR-135a Targets SMAD2 to Promote Osteosarcoma Proliferation and Migration
CN109224076B (en) Gene miR-140-3P related to lung cancer diagnosis and treatment, and mimics and application thereof
CN110777204B (en) Application of MAFG-AS1 knock-out reagent in preparation of medicines for treating bladder cancer
Yao et al. Bioinformatics analysis of prognosis-related genes and expression of CXCL8 in colorectal cancer
CN114703287B (en) Application of CXorf56 gene in treating triple negative breast cancer
CN116676392B (en) Application of USP10-MOF-ANXA2 signal path as drug target in preparation of drug for treating esophageal squamous cell carcinoma

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
WW01 Invention patent application withdrawn after publication
WW01 Invention patent application withdrawn after publication

Application publication date: 20190712