JP4751407B2 - ヒトt細胞白血病ウイルス感染あるいは成人t細胞白血病発症の検出法、その診断薬、ならびにその予防および治療の為の薬剤 - Google Patents
ヒトt細胞白血病ウイルス感染あるいは成人t細胞白血病発症の検出法、その診断薬、ならびにその予防および治療の為の薬剤 Download PDFInfo
- Publication number
- JP4751407B2 JP4751407B2 JP2008031855A JP2008031855A JP4751407B2 JP 4751407 B2 JP4751407 B2 JP 4751407B2 JP 2008031855 A JP2008031855 A JP 2008031855A JP 2008031855 A JP2008031855 A JP 2008031855A JP 4751407 B2 JP4751407 B2 JP 4751407B2
- Authority
- JP
- Japan
- Prior art keywords
- ndrg2
- atl
- cells
- cell
- htlv
- 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.)
- Expired - Fee Related
Links
- 238000001514 detection method Methods 0.000 title claims description 18
- 230000002265 prevention Effects 0.000 title claims description 15
- 239000003814 drug Substances 0.000 title description 9
- 229940079593 drug Drugs 0.000 title description 8
- 229940039227 diagnostic agent Drugs 0.000 title description 5
- 239000000032 diagnostic agent Substances 0.000 title description 5
- 208000000389 T-cell leukemia Diseases 0.000 title description 4
- 208000009746 Adult T-Cell Leukemia-Lymphoma Diseases 0.000 title description 2
- 208000016683 Adult T-cell leukemia/lymphoma Diseases 0.000 title description 2
- 208000028530 T-cell lymphoblastic leukemia/lymphoma Diseases 0.000 title description 2
- 201000006966 adult T-cell leukemia Diseases 0.000 title description 2
- 230000009385 viral infection Effects 0.000 title 1
- 101000995300 Homo sapiens Protein NDRG2 Proteins 0.000 claims description 133
- 102100034436 Protein NDRG2 Human genes 0.000 claims description 133
- 230000014509 gene expression Effects 0.000 claims description 80
- 238000000034 method Methods 0.000 claims description 58
- 238000007069 methylation reaction Methods 0.000 claims description 48
- 230000011987 methylation Effects 0.000 claims description 46
- 208000015181 infectious disease Diseases 0.000 claims description 21
- 108091033319 polynucleotide Proteins 0.000 claims description 12
- 102000040430 polynucleotide Human genes 0.000 claims description 12
- 239000002157 polynucleotide Substances 0.000 claims description 12
- 239000003795 chemical substances by application Substances 0.000 claims description 7
- 241000714260 Human T-lymphotropic virus 1 Species 0.000 claims description 6
- 108010033276 Peptide Fragments Proteins 0.000 claims description 3
- 102000007079 Peptide Fragments Human genes 0.000 claims description 3
- 210000004027 cell Anatomy 0.000 description 169
- 101150006362 atl gene Proteins 0.000 description 109
- 239000013615 primer Substances 0.000 description 46
- 108090000623 proteins and genes Proteins 0.000 description 34
- 101150096352 Ndrg2 gene Proteins 0.000 description 27
- 108090000765 processed proteins & peptides Proteins 0.000 description 22
- 108020004414 DNA Proteins 0.000 description 17
- 238000006243 chemical reaction Methods 0.000 description 15
- 230000001154 acute effect Effects 0.000 description 14
- 229920001184 polypeptide Polymers 0.000 description 14
- 102000004196 processed proteins & peptides Human genes 0.000 description 14
- 108700039691 Genetic Promoter Regions Proteins 0.000 description 13
- 210000001744 T-lymphocyte Anatomy 0.000 description 13
- 108020004999 messenger RNA Proteins 0.000 description 13
- 102000004169 proteins and genes Human genes 0.000 description 13
- 239000002299 complementary DNA Substances 0.000 description 12
- 239000012634 fragment Substances 0.000 description 12
- 238000004458 analytical method Methods 0.000 description 11
- 208000032839 leukemia Diseases 0.000 description 11
- 230000002441 reversible effect Effects 0.000 description 11
- 230000007067 DNA methylation Effects 0.000 description 10
- 125000003729 nucleotide group Chemical group 0.000 description 10
- 150000003839 salts Chemical class 0.000 description 10
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 9
- 239000000872 buffer Substances 0.000 description 9
- 239000002773 nucleotide Substances 0.000 description 9
- 210000004940 nucleus Anatomy 0.000 description 9
- 210000002966 serum Anatomy 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 239000013598 vector Substances 0.000 description 9
- LSNNMFCWUKXFEE-UHFFFAOYSA-M Bisulfite Chemical compound OS([O-])=O LSNNMFCWUKXFEE-UHFFFAOYSA-M 0.000 description 8
- 239000013604 expression vector Substances 0.000 description 8
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 7
- 206010028980 Neoplasm Diseases 0.000 description 7
- 230000008859 change Effects 0.000 description 7
- 238000000338 in vitro Methods 0.000 description 7
- 238000001727 in vivo Methods 0.000 description 7
- 230000010473 stable expression Effects 0.000 description 7
- 108010085238 Actins Proteins 0.000 description 6
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 241000283973 Oryctolagus cuniculus Species 0.000 description 6
- 241000700605 Viruses Species 0.000 description 6
- 239000000427 antigen Substances 0.000 description 6
- 108091007433 antigens Proteins 0.000 description 6
- 102000036639 antigens Human genes 0.000 description 6
- 230000036961 partial effect Effects 0.000 description 6
- 238000011144 upstream manufacturing Methods 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 5
- 241000699670 Mus sp. Species 0.000 description 5
- 108091034117 Oligonucleotide Proteins 0.000 description 5
- 210000004369 blood Anatomy 0.000 description 5
- 239000008280 blood Substances 0.000 description 5
- 201000011510 cancer Diseases 0.000 description 5
- OPTASPLRGRRNAP-UHFFFAOYSA-N cytosine Chemical compound NC=1C=CNC(=O)N=1 OPTASPLRGRRNAP-UHFFFAOYSA-N 0.000 description 5
- 230000003247 decreasing effect Effects 0.000 description 5
- 238000011161 development Methods 0.000 description 5
- 230000018109 developmental process Effects 0.000 description 5
- 239000011325 microbead Substances 0.000 description 5
- 210000005105 peripheral blood lymphocyte Anatomy 0.000 description 5
- 239000000047 product Substances 0.000 description 5
- 230000035755 proliferation Effects 0.000 description 5
- 239000000523 sample Substances 0.000 description 5
- 230000004083 survival effect Effects 0.000 description 5
- 241000283707 Capra Species 0.000 description 4
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 4
- 230000004544 DNA amplification Effects 0.000 description 4
- 108010001336 Horseradish Peroxidase Proteins 0.000 description 4
- 108050001500 NDRG Proteins 0.000 description 4
- ISAKRJDGNUQOIC-UHFFFAOYSA-N Uracil Chemical compound O=C1C=CNC(=O)N1 ISAKRJDGNUQOIC-UHFFFAOYSA-N 0.000 description 4
- 125000000539 amino acid group Chemical group 0.000 description 4
- 230000003321 amplification Effects 0.000 description 4
- 230000015572 biosynthetic process Effects 0.000 description 4
- 125000003178 carboxy group Chemical group [H]OC(*)=O 0.000 description 4
- 230000004927 fusion Effects 0.000 description 4
- 230000012010 growth Effects 0.000 description 4
- 230000004048 modification Effects 0.000 description 4
- 238000012986 modification Methods 0.000 description 4
- 238000003199 nucleic acid amplification method Methods 0.000 description 4
- 210000005259 peripheral blood Anatomy 0.000 description 4
- 239000011886 peripheral blood Substances 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- 230000003612 virological effect Effects 0.000 description 4
- 238000005406 washing Methods 0.000 description 4
- 238000001262 western blot Methods 0.000 description 4
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 3
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 3
- 210000004366 CD4-positive T-lymphocyte Anatomy 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 108091060211 Expressed sequence tag Proteins 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 108010002350 Interleukin-2 Proteins 0.000 description 3
- MUBZPKHOEPUJKR-UHFFFAOYSA-N Oxalic acid Chemical compound OC(=O)C(O)=O MUBZPKHOEPUJKR-UHFFFAOYSA-N 0.000 description 3
- RTKIYFITIVXBLE-UHFFFAOYSA-N Trichostatin A Natural products ONC(=O)C=CC(C)=CC(C)C(=O)C1=CC=C(N(C)C)C=C1 RTKIYFITIVXBLE-UHFFFAOYSA-N 0.000 description 3
- 239000002246 antineoplastic agent Substances 0.000 description 3
- 230000010261 cell growth Effects 0.000 description 3
- 230000000295 complement effect Effects 0.000 description 3
- 238000003745 diagnosis Methods 0.000 description 3
- 210000004408 hybridoma Anatomy 0.000 description 3
- 230000003053 immunization Effects 0.000 description 3
- 238000003018 immunoassay Methods 0.000 description 3
- 150000002632 lipids Chemical class 0.000 description 3
- 230000007246 mechanism Effects 0.000 description 3
- 239000000203 mixture Substances 0.000 description 3
- 238000011002 quantification Methods 0.000 description 3
- 238000003753 real-time PCR Methods 0.000 description 3
- 238000011084 recovery Methods 0.000 description 3
- 230000002829 reductive effect Effects 0.000 description 3
- 230000010076 replication Effects 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- RTKIYFITIVXBLE-QEQCGCAPSA-N trichostatin A Chemical compound ONC(=O)/C=C/C(/C)=C/[C@@H](C)C(=O)C1=CC=C(N(C)C)C=C1 RTKIYFITIVXBLE-QEQCGCAPSA-N 0.000 description 3
- 239000013603 viral vector Substances 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- FWMNVWWHGCHHJJ-SKKKGAJSSA-N 4-amino-1-[(2r)-6-amino-2-[[(2r)-2-[[(2r)-2-[[(2r)-2-amino-3-phenylpropanoyl]amino]-3-phenylpropanoyl]amino]-4-methylpentanoyl]amino]hexanoyl]piperidine-4-carboxylic acid Chemical compound C([C@H](C(=O)N[C@H](CC(C)C)C(=O)N[C@H](CCCCN)C(=O)N1CCC(N)(CC1)C(O)=O)NC(=O)[C@H](N)CC=1C=CC=CC=1)C1=CC=CC=C1 FWMNVWWHGCHHJJ-SKKKGAJSSA-N 0.000 description 2
- XAUDJQYHKZQPEU-KVQBGUIXSA-N 5-aza-2'-deoxycytidine Chemical compound O=C1N=C(N)N=CN1[C@@H]1O[C@H](CO)[C@@H](O)C1 XAUDJQYHKZQPEU-KVQBGUIXSA-N 0.000 description 2
- HRPVXLWXLXDGHG-UHFFFAOYSA-N Acrylamide Chemical compound NC(=O)C=C HRPVXLWXLXDGHG-UHFFFAOYSA-N 0.000 description 2
- 102000007469 Actins Human genes 0.000 description 2
- 206010000830 Acute leukaemia Diseases 0.000 description 2
- ZAINTDRBUHCDPZ-UHFFFAOYSA-M Alexa Fluor 546 Chemical compound [H+].[Na+].CC1CC(C)(C)NC(C(=C2OC3=C(C4=NC(C)(C)CC(C)C4=CC3=3)S([O-])(=O)=O)S([O-])(=O)=O)=C1C=C2C=3C(C(=C(Cl)C=1Cl)C(O)=O)=C(Cl)C=1SCC(=O)NCCCCCC(=O)ON1C(=O)CCC1=O ZAINTDRBUHCDPZ-UHFFFAOYSA-M 0.000 description 2
- QGZKDVFQNNGYKY-UHFFFAOYSA-N Ammonia Chemical compound N QGZKDVFQNNGYKY-UHFFFAOYSA-N 0.000 description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 2
- 102000003826 Chemokine CCL17 Human genes 0.000 description 2
- 108010082169 Chemokine CCL17 Proteins 0.000 description 2
- 102000006433 Chemokine CCL22 Human genes 0.000 description 2
- 108010083701 Chemokine CCL22 Proteins 0.000 description 2
- 238000000116 DAPI staining Methods 0.000 description 2
- 241000282412 Homo Species 0.000 description 2
- 241000725303 Human immunodeficiency virus Species 0.000 description 2
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 description 2
- QIGBRXMKCJKVMJ-UHFFFAOYSA-N Hydroquinone Chemical compound OC1=CC=C(O)C=C1 QIGBRXMKCJKVMJ-UHFFFAOYSA-N 0.000 description 2
- XEEYBQQBJWHFJM-UHFFFAOYSA-N Iron Chemical compound [Fe] XEEYBQQBJWHFJM-UHFFFAOYSA-N 0.000 description 2
- 241000714177 Murine leukemia virus Species 0.000 description 2
- 238000000636 Northern blotting Methods 0.000 description 2
- 238000009004 PCR Kit Methods 0.000 description 2
- 241000711504 Paramyxoviridae Species 0.000 description 2
- NQRYJNQNLNOLGT-UHFFFAOYSA-N Piperidine Chemical compound C1CCNCC1 NQRYJNQNLNOLGT-UHFFFAOYSA-N 0.000 description 2
- 238000011529 RT qPCR Methods 0.000 description 2
- 241000700159 Rattus Species 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 241000713311 Simian immunodeficiency virus Species 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 2
- 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 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 125000002252 acyl group Chemical group 0.000 description 2
- 238000000137 annealing Methods 0.000 description 2
- 238000013459 approach Methods 0.000 description 2
- 239000011324 bead Substances 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000000903 blocking effect Effects 0.000 description 2
- 210000001124 body fluid Anatomy 0.000 description 2
- 239000010839 body fluid Substances 0.000 description 2
- 239000001569 carbon dioxide Substances 0.000 description 2
- 229910002092 carbon dioxide Inorganic materials 0.000 description 2
- 230000004663 cell proliferation Effects 0.000 description 2
- 238000010367 cloning Methods 0.000 description 2
- 230000002596 correlated effect Effects 0.000 description 2
- 230000000875 corresponding effect Effects 0.000 description 2
- 238000004132 cross linking Methods 0.000 description 2
- 229940104302 cytosine Drugs 0.000 description 2
- 230000034994 death Effects 0.000 description 2
- 238000010520 demethylation reaction Methods 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 238000013399 early diagnosis Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 238000012869 ethanol precipitation Methods 0.000 description 2
- 238000010195 expression analysis Methods 0.000 description 2
- 239000012091 fetal bovine serum Substances 0.000 description 2
- 238000010353 genetic engineering Methods 0.000 description 2
- 238000009396 hybridization Methods 0.000 description 2
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 2
- 238000002649 immunization Methods 0.000 description 2
- 238000012744 immunostaining Methods 0.000 description 2
- 239000003112 inhibitor Substances 0.000 description 2
- 210000004698 lymphocyte Anatomy 0.000 description 2
- 238000004519 manufacturing process Methods 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000007855 methylation-specific PCR Methods 0.000 description 2
- 230000035772 mutation Effects 0.000 description 2
- 238000010647 peptide synthesis reaction Methods 0.000 description 2
- 238000003752 polymerase chain reaction Methods 0.000 description 2
- 230000001177 retroviral effect Effects 0.000 description 2
- 238000010839 reverse transcription Methods 0.000 description 2
- 238000003757 reverse transcription PCR Methods 0.000 description 2
- 235000002020 sage Nutrition 0.000 description 2
- 238000000926 separation method Methods 0.000 description 2
- 239000007790 solid phase Substances 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012546 transfer Methods 0.000 description 2
- 230000009466 transformation Effects 0.000 description 2
- 238000002054 transplantation Methods 0.000 description 2
- 229940035893 uracil Drugs 0.000 description 2
- 125000003088 (fluoren-9-ylmethoxy)carbonyl group Chemical group 0.000 description 1
- FWBHETKCLVMNFS-UHFFFAOYSA-N 4',6-Diamino-2-phenylindol Chemical compound C1=CC(C(=N)N)=CC=C1C1=CC2=CC=C(C(N)=N)C=C2N1 FWBHETKCLVMNFS-UHFFFAOYSA-N 0.000 description 1
- ODHCTXKNWHHXJC-VKHMYHEASA-N 5-oxo-L-proline Chemical compound OC(=O)[C@@H]1CCC(=O)N1 ODHCTXKNWHHXJC-VKHMYHEASA-N 0.000 description 1
- 101150106774 9 gene Proteins 0.000 description 1
- 230000005730 ADP ribosylation Effects 0.000 description 1
- 241000701242 Adenoviridae Species 0.000 description 1
- 229920000936 Agarose Polymers 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 1
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 1
- QGZKDVFQNNGYKY-UHFFFAOYSA-O Ammonium Chemical compound [NH4+] QGZKDVFQNNGYKY-UHFFFAOYSA-O 0.000 description 1
- 101100003227 Arabidopsis thaliana ATL38 gene Proteins 0.000 description 1
- 101100217578 Arabidopsis thaliana ATL66 gene Proteins 0.000 description 1
- 239000004475 Arginine Substances 0.000 description 1
- 102100024222 B-lymphocyte antigen CD19 Human genes 0.000 description 1
- 244000063299 Bacillus subtilis Species 0.000 description 1
- 235000014469 Bacillus subtilis Nutrition 0.000 description 1
- 241000701412 Baculoviridae Species 0.000 description 1
- 101000800130 Bos taurus Thyroglobulin Proteins 0.000 description 1
- 101710149863 C-C chemokine receptor type 4 Proteins 0.000 description 1
- 102100032976 CCR4-NOT transcription complex subunit 6 Human genes 0.000 description 1
- 102000049320 CD36 Human genes 0.000 description 1
- 108010045374 CD36 Antigens Proteins 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 108010078791 Carrier Proteins Proteins 0.000 description 1
- 102000014914 Carrier Proteins Human genes 0.000 description 1
- 102100039496 Choline transporter-like protein 4 Human genes 0.000 description 1
- 241000699800 Cricetinae Species 0.000 description 1
- LEVWYRKDKASIDU-QWWZWVQMSA-N D-cystine Chemical compound OC(=O)[C@H](N)CSSC[C@@H](N)C(O)=O LEVWYRKDKASIDU-QWWZWVQMSA-N 0.000 description 1
- 230000035131 DNA demethylation Effects 0.000 description 1
- 239000003155 DNA primer Substances 0.000 description 1
- 241000702421 Dependoparvovirus Species 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000713813 Gibbon ape leukemia virus Species 0.000 description 1
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Natural products NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 description 1
- 239000004471 Glycine Substances 0.000 description 1
- 102100035716 Glycophorin-A Human genes 0.000 description 1
- 108091005250 Glycophorins Proteins 0.000 description 1
- 206010019842 Hepatomegaly Diseases 0.000 description 1
- 241000700586 Herpesviridae Species 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 101000980825 Homo sapiens B-lymphocyte antigen CD19 Proteins 0.000 description 1
- 101100005713 Homo sapiens CD4 gene Proteins 0.000 description 1
- 101000889282 Homo sapiens Choline transporter-like protein 4 Proteins 0.000 description 1
- 101000998120 Homo sapiens Interleukin-3 receptor subunit alpha Proteins 0.000 description 1
- 101000917858 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-A Proteins 0.000 description 1
- 101000917839 Homo sapiens Low affinity immunoglobulin gamma Fc region receptor III-B Proteins 0.000 description 1
- 101000946889 Homo sapiens Monocyte differentiation antigen CD14 Proteins 0.000 description 1
- 101100133104 Homo sapiens NDRG2 gene Proteins 0.000 description 1
- 101000581981 Homo sapiens Neural cell adhesion molecule 1 Proteins 0.000 description 1
- 101000946843 Homo sapiens T-cell surface glycoprotein CD8 alpha chain Proteins 0.000 description 1
- 241000598436 Human T-cell lymphotropic virus Species 0.000 description 1
- 241000598171 Human adenovirus sp. Species 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- 108060003951 Immunoglobulin Proteins 0.000 description 1
- 102000001706 Immunoglobulin Fab Fragments Human genes 0.000 description 1
- 108010054477 Immunoglobulin Fab Fragments Proteins 0.000 description 1
- 102000017727 Immunoglobulin Variable Region Human genes 0.000 description 1
- 108010067060 Immunoglobulin Variable Region Proteins 0.000 description 1
- 102100033493 Interleukin-3 receptor subunit alpha Human genes 0.000 description 1
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 1
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 description 1
- 102100029185 Low affinity immunoglobulin gamma Fc region receptor III-B Human genes 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 102100035877 Monocyte differentiation antigen CD14 Human genes 0.000 description 1
- 241000711408 Murine respirovirus Species 0.000 description 1
- 241000237536 Mytilus edulis Species 0.000 description 1
- 108700026495 N-Myc Proto-Oncogene Proteins 0.000 description 1
- 102100030124 N-myc proto-oncogene protein Human genes 0.000 description 1
- 102000011324 NDRG Human genes 0.000 description 1
- 102100027347 Neural cell adhesion molecule 1 Human genes 0.000 description 1
- 239000002033 PVDF binder Substances 0.000 description 1
- 206010061902 Pancreatic neoplasm Diseases 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- 241000701945 Parvoviridae Species 0.000 description 1
- 206010035226 Plasma cell myeloma Diseases 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 229920001213 Polysorbate 20 Polymers 0.000 description 1
- 241000700625 Poxviridae Species 0.000 description 1
- 108010026552 Proteome Proteins 0.000 description 1
- 239000012980 RPMI-1640 medium Substances 0.000 description 1
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 1
- DWAQJAXMDSEUJJ-UHFFFAOYSA-M Sodium bisulfite Chemical compound [Na+].OS([O-])=O DWAQJAXMDSEUJJ-UHFFFAOYSA-M 0.000 description 1
- LSNNMFCWUKXFEE-UHFFFAOYSA-N Sulfurous acid Chemical group OS(O)=O LSNNMFCWUKXFEE-UHFFFAOYSA-N 0.000 description 1
- 102100034922 T-cell surface glycoprotein CD8 alpha chain Human genes 0.000 description 1
- -1 TCRγ / σ Proteins 0.000 description 1
- GSEJCLTVZPLZKY-UHFFFAOYSA-N Triethanolamine Chemical compound OCCN(CCO)CCO GSEJCLTVZPLZKY-UHFFFAOYSA-N 0.000 description 1
- 239000007984 Tris EDTA buffer Substances 0.000 description 1
- GLNADSQYFUSGOU-GPTZEZBUSA-J Trypan blue Chemical compound [Na+].[Na+].[Na+].[Na+].C1=C(S([O-])(=O)=O)C=C2C=C(S([O-])(=O)=O)C(/N=N/C3=CC=C(C=C3C)C=3C=C(C(=CC=3)\N=N\C=3C(=CC4=CC(=CC(N)=C4C=3O)S([O-])(=O)=O)S([O-])(=O)=O)C)=C(O)C2=C1N GLNADSQYFUSGOU-GPTZEZBUSA-J 0.000 description 1
- 102000044209 Tumor Suppressor Genes Human genes 0.000 description 1
- 108700025716 Tumor Suppressor Genes Proteins 0.000 description 1
- 235000010724 Wisteria floribunda Nutrition 0.000 description 1
- HCHKCACWOHOZIP-UHFFFAOYSA-N Zinc Chemical compound [Zn] HCHKCACWOHOZIP-UHFFFAOYSA-N 0.000 description 1
- 230000005856 abnormality Effects 0.000 description 1
- 230000035508 accumulation Effects 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000021736 acetylation Effects 0.000 description 1
- 238000006640 acetylation reaction Methods 0.000 description 1
- 150000007513 acids Chemical class 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000010933 acylation Effects 0.000 description 1
- 238000005917 acylation reaction Methods 0.000 description 1
- 108700010877 adenoviridae proteins Proteins 0.000 description 1
- 125000001931 aliphatic group Chemical group 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- 102000005840 alpha-Galactosidase Human genes 0.000 description 1
- 108010030291 alpha-Galactosidase Proteins 0.000 description 1
- 230000009435 amidation Effects 0.000 description 1
- 238000007112 amidation reaction Methods 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 150000001412 amines Chemical class 0.000 description 1
- 230000006229 amino acid addition Effects 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 125000003277 amino group Chemical group 0.000 description 1
- 229910021529 ammonia Inorganic materials 0.000 description 1
- 230000003302 anti-idiotype Effects 0.000 description 1
- 210000000628 antibody-producing cell Anatomy 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 1
- 230000037429 base substitution Effects 0.000 description 1
- 230000027455 binding Effects 0.000 description 1
- 238000012742 biochemical analysis Methods 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 238000001369 bisulfite sequencing Methods 0.000 description 1
- 210000000601 blood cell Anatomy 0.000 description 1
- 238000006664 bond formation reaction Methods 0.000 description 1
- 239000012888 bovine serum Substances 0.000 description 1
- 210000004958 brain cell Anatomy 0.000 description 1
- 210000000481 breast Anatomy 0.000 description 1
- UDSAIICHUKSCKT-UHFFFAOYSA-N bromophenol blue Chemical compound C1=C(Br)C(O)=C(Br)C=C1C1(C=2C=C(Br)C(O)=C(Br)C=2)C2=CC=CC=C2S(=O)(=O)O1 UDSAIICHUKSCKT-UHFFFAOYSA-N 0.000 description 1
- 210000004899 c-terminal region Anatomy 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000000969 carrier Substances 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 210000003855 cell nucleus Anatomy 0.000 description 1
- 230000030570 cellular localization Effects 0.000 description 1
- 210000000349 chromosome Anatomy 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 230000003750 conditioning effect Effects 0.000 description 1
- 238000012136 culture method Methods 0.000 description 1
- 229960003067 cystine Drugs 0.000 description 1
- 210000000805 cytoplasm Anatomy 0.000 description 1
- 230000002950 deficient Effects 0.000 description 1
- 230000003111 delayed effect Effects 0.000 description 1
- 230000017858 demethylation Effects 0.000 description 1
- 210000004443 dendritic cell Anatomy 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 238000003113 dilution method Methods 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- 238000012377 drug delivery Methods 0.000 description 1
- 238000001962 electrophoresis Methods 0.000 description 1
- 238000004520 electroporation Methods 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 230000001973 epigenetic effect Effects 0.000 description 1
- 210000003527 eukaryotic cell Anatomy 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 210000004700 fetal blood Anatomy 0.000 description 1
- 230000022244 formylation Effects 0.000 description 1
- 238000006170 formylation reaction Methods 0.000 description 1
- 125000000524 functional group Chemical group 0.000 description 1
- 108020001507 fusion proteins Proteins 0.000 description 1
- 102000037865 fusion proteins Human genes 0.000 description 1
- 230000006251 gamma-carboxylation Effects 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 230000004547 gene signature Effects 0.000 description 1
- 238000001415 gene therapy Methods 0.000 description 1
- 210000004907 gland Anatomy 0.000 description 1
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 description 1
- 125000003827 glycol group Chemical group 0.000 description 1
- 230000013595 glycosylation Effects 0.000 description 1
- 238000006206 glycosylation reaction Methods 0.000 description 1
- 125000003630 glycyl group Chemical group [H]N([H])C([H])([H])C(*)=O 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 150000003278 haem Chemical group 0.000 description 1
- 108060003552 hemocyanin Proteins 0.000 description 1
- 229940121372 histone deacetylase inhibitor Drugs 0.000 description 1
- 239000003276 histone deacetylase inhibitor Substances 0.000 description 1
- 230000006197 histone deacetylation Effects 0.000 description 1
- 210000005104 human peripheral blood lymphocyte Anatomy 0.000 description 1
- 230000033444 hydroxylation Effects 0.000 description 1
- 238000005805 hydroxylation reaction Methods 0.000 description 1
- 230000006607 hypermethylation Effects 0.000 description 1
- 238000010191 image analysis Methods 0.000 description 1
- 230000001900 immune effect Effects 0.000 description 1
- 230000008105 immune reaction Effects 0.000 description 1
- 102000018358 immunoglobulin Human genes 0.000 description 1
- 230000016784 immunoglobulin production Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000026045 iodination Effects 0.000 description 1
- 238000006192 iodination reaction Methods 0.000 description 1
- 229910052742 iron Inorganic materials 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 210000003292 kidney cell Anatomy 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000004576 lipid-binding Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 238000004895 liquid chromatography mass spectrometry Methods 0.000 description 1
- 201000007270 liver cancer Diseases 0.000 description 1
- 208000014018 liver neoplasm Diseases 0.000 description 1
- 230000004807 localization Effects 0.000 description 1
- 210000001165 lymph node Anatomy 0.000 description 1
- 238000002826 magnetic-activated cell sorting Methods 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 208000015486 malignant pancreatic neoplasm Diseases 0.000 description 1
- 210000004962 mammalian cell Anatomy 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 206010027191 meningioma Diseases 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 238000000520 microinjection Methods 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 239000003068 molecular probe Substances 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 210000005087 mononuclear cell Anatomy 0.000 description 1
- 239000012120 mounting media Substances 0.000 description 1
- 210000000663 muscle cell Anatomy 0.000 description 1
- 235000020638 mussel Nutrition 0.000 description 1
- 201000000050 myeloid neoplasm Diseases 0.000 description 1
- 230000007498 myristoylation Effects 0.000 description 1
- ZIUHHBKFKCYYJD-UHFFFAOYSA-N n,n'-methylenebisacrylamide Chemical compound C=CC(=O)NCNC(=O)C=C ZIUHHBKFKCYYJD-UHFFFAOYSA-N 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 150000007530 organic bases Chemical class 0.000 description 1
- 235000006408 oxalic acid Nutrition 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 201000002528 pancreatic cancer Diseases 0.000 description 1
- 208000008443 pancreatic carcinoma Diseases 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000003950 pathogenic mechanism Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 150000003905 phosphatidylinositols Chemical class 0.000 description 1
- 230000026731 phosphorylation Effects 0.000 description 1
- 238000006366 phosphorylation reaction Methods 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 239000000256 polyoxyethylene sorbitan monolaurate Substances 0.000 description 1
- 235000010486 polyoxyethylene sorbitan monolaurate Nutrition 0.000 description 1
- 229920002981 polyvinylidene fluoride Polymers 0.000 description 1
- 238000010837 poor prognosis Methods 0.000 description 1
- 230000013823 prenylation Effects 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 150000003141 primary amines Chemical class 0.000 description 1
- MFDFERRIHVXMIY-UHFFFAOYSA-N procaine Chemical compound CCN(CC)CCOC(=O)C1=CC=C(N)C=C1 MFDFERRIHVXMIY-UHFFFAOYSA-N 0.000 description 1
- 229960004919 procaine Drugs 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 238000004393 prognosis Methods 0.000 description 1
- 210000001236 prokaryotic cell Anatomy 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000026447 protein localization Effects 0.000 description 1
- 230000002797 proteolythic effect Effects 0.000 description 1
- 229940043131 pyroglutamate Drugs 0.000 description 1
- 238000004445 quantitative analysis Methods 0.000 description 1
- 230000006340 racemization Effects 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012827 research and development Methods 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 238000007363 ring formation reaction Methods 0.000 description 1
- 150000003335 secondary amines Chemical class 0.000 description 1
- 238000013207 serial dilution Methods 0.000 description 1
- 125000003607 serino group Chemical group [H]N([H])[C@]([H])(C(=O)[*])C(O[H])([H])[H] 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 206010040882 skin lesion Diseases 0.000 description 1
- 231100000444 skin lesion Toxicity 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- 235000010267 sodium hydrogen sulphite Nutrition 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 230000006641 stabilisation Effects 0.000 description 1
- 238000011105 stabilization Methods 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 230000000638 stimulation Effects 0.000 description 1
- 238000003860 storage Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000019635 sulfation Effects 0.000 description 1
- 238000005670 sulfation reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001629 suppression Effects 0.000 description 1
- 238000001356 surgical procedure Methods 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 238000002626 targeted therapy Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 125000000341 threoninyl group Chemical group [H]OC([H])(C([H])([H])[H])C([H])(N([H])[H])C(*)=O 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 238000003151 transfection method Methods 0.000 description 1
- 150000003852 triazoles Chemical class 0.000 description 1
- 210000004881 tumor cell Anatomy 0.000 description 1
- 230000005740 tumor formation Effects 0.000 description 1
- 238000000539 two dimensional gel electrophoresis Methods 0.000 description 1
- 241000701161 unidentified adenovirus Species 0.000 description 1
- 241001430294 unidentified retrovirus Species 0.000 description 1
- 238000012800 visualization Methods 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000011701 zinc Substances 0.000 description 1
- 229910052725 zinc Inorganic materials 0.000 description 1
Images
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Description
Ishida T, Utsunomiya A, et al. Clin. Cancer Res. 9: 3625-3634, 2003
被験者由来の細胞中のNDRG2の発現レベルを測定し、コントロール細胞中のNDRG2の発現レベルと比較する工程;および
該比較において、被験者由来の細胞中のNDRG2の発現レベルが、コントロール細胞中のNDRG2の発現レベルより低下していることを指標として、被験者がHTLV−1の感染者および/またはATL発症者であると決定する工程
を含む、検出方法、を提供する。
被験者由来の細胞中のNDRG2プロモーターのメチル化を測定する工程;およびメチル化がされていることを指標として、被験者がHTLV−1の感染者および/またはATL発症者であると決定する工程、を含む検出方法を提供する。
(健常人由来CD4陽性細胞及び患者検体)
HTLV−1に感染していない健常人、もしくは、ATL患者ヘパリン加末梢血をHistopaque (シグマアルドリッチ)による比重遠心法を用いて単核球を遠心分離した。急性型ATL患者では、末梢血ATL分画が90%異常の検体を解析に用いた。また健常人のCD4陽性細胞はCD4+ T細胞 アイソレーションキットII (Miltenyi Biotec)によりCD4+T細胞以外の細胞をCD8、CD14、CD 16、CD19、CD36、CD56、CD123、TCRγ/σ、Glycophorin Aに対するビオチン標識抗体カクテルによりAuto MACS (Miltenyi Biotec)を用いて分離した。
実験に用いた細胞株は次の通りである。HTLV−1非感染T細胞リンパ性白血病(T−ALL)細胞株としてJurkat、MOLT4、MKB-1、KAWAI、HTLV−1感染細胞株としてHUT102、MT2、IL2非依存性ATL細胞株ED-40515(-)、Su9T-01、S1T、IL2依存性ATL細胞株KOB、KK1、SO4を、それぞれIL−2 (50 U/ml)添加もしくは無添加のfetal bovine serum (FBS)を10%添加したRPMI1640培養液で37 度、5%二酸化炭素下で培養した。
染色体14q11領域のNDRG2遺伝子近傍の遺伝子9個について、T−ALL(Tリンパ性急性白血病)細胞株2株(Jurkat、MOLT4)とHTLV−1感染細胞株3株(HUT102、MT2、OMT)及びATL細胞株6株(ED-40515(-)、Su9T-01、S1T、KOB、KK1、SO4)を用いて半定量的RT−PCR法を用いて遺伝子発現を検討した。
(1)抗体
NDRG2タンパク質発現を同定する目的で、ウサギ抗ヒトNDRG2抗体を調製した。NDRG2ペプチド(NDRG2 B型 (NP_057334 ) 17−31 アミノ酸残基 PGQTPEAAKTHSVET(配列表の配列番号11)を合成し、家兎に免疫、さらに免疫後血清よりIgG分画を精製し、ペプチド固層カラムを用いた精製を行ない作製した。その他の抗体としてマウス抗β-アクチン(AC-15) モノクローナル抗体(A5441)、ウサギ抗FLAGポリクローナル抗体 (F7425)、マウス抗FLAG (M2)モノクローナル抗体 (F1804) (シグマアルドリッチ)を使用した。また二次抗体としてワサビペルオキシダーゼ(HRP)標識抗マウス IgG 抗体(NA931V) (GE Healthcare) ワサビペルオキシダーゼ標識抗ウサギ IgG 抗体(P0399) (Dako Cytomation)、及びAlexaFluor 546標識抗ヤギ抗マウスIgG (重鎖、軽鎖)抗体 、フルオロセン標識ヤギ抗ウサギIgG (重鎖、軽鎖)抗体(Molecular Probe)を使用した。
各種白血病細胞並びに健常人CD4陽性Tリンパ球、急性型ATL白血病細胞を用いてNDRG2タンパク質発現をβ−アクチンタンパク質発現を標準コントロールとしてウエスタン法により検討した。
NDRG2遺伝子の発現抑制機構の解析
まずDNAメチル化阻害剤である5-aza-dC (5-aza-2’-deoxycytidine)もしくはヒストン脱アセチル化酵素阻害剤であるTSA (Trichostatin A)をATL細胞株に作用させ遺伝子発現の変化を検討した。コントロールとしてT−ALL (Tリンパ性急性白血病)細胞株4株(Jurkat、MOLT4、MKB-1、KAWAI )、HTLV−1感染細胞株2株(HUT102、MT2)、ATL細胞株6株(ED-40515(-)、Su9T-01、S1T, KOB、KK1、SO4 )を用いた。その結果T−ALL細胞株では発現の変化が見られなかったがHTLV−1感染細胞株及びATL細胞株全8株中、5-aza-dc処理により7株(HUT102、ED-40515(-)、Su9T-01、S1T, KOB、KK1、SO4)、TSA処理により7株(MT2、ED-40515(-)、Su9T-01、S1T, KOB、KK1、SO4)において発現の回復が認められた(図5)。従ってNDRG2遺伝子発現低下の原因の一つとしてエピジェネティックなゲノム異常が関係している可能性が示唆した。
ヒトNDRG2遺伝子は、MOLT4細胞株より抽出したtotal RNAを鋳型としてAMV-RT RNA PCR Kit ver3.1 (タカラバイオ) 中のオリゴdT プライマーを用いてcDNAを作成した。このcDNAを鋳型として、NDRG2-5’-HindIIIプライマー 5’-CCCAAGCTTATGGCGGAGCTGCAGGAGGTGC-3’(配列表の配列番号12)及びNDRG2-3’-EcoRI プライマー 5’-GCGAATTCTCAACAGGAGACCTCCATG-3’(配列表の配列番号13)を用いてNDRG2 B型 (NM_016250)として PCR 30サイクル(98 度 10 秒、55 度 30 秒、72 度 1 分15 秒) で増幅した。PCR産物をTArget Clone (TOYOBO)中のpTA2ベクターに導入した (NDRG2/pTA2)。NDRG2 cDNA産物は塩基配列を確認した後、pCMV26ベクター(シグマアルドリッチ)のCMVプロモーター下流Hind III/EcoRI領域に3×FLAG tag融合タンパク質として導入した(pCMV26/NDRG2)。
ATL細胞株KOB、KK1、Su9T-01細胞1 x 107 個に20 μgのpCMV26/NDRG2もしくはpCMV26/mockをGENE PLUSER II (Bio-Rad, Hercules, CA, USA)を用いて220 V、950 ・Fの条件で遺伝子導入を行なった。導入2 日後、培養液中にG418を800 μg/ml添加し、その1週間後限界希釈法によって細胞の株化を行なった。安定発現株樹立後はそれぞれの培地にG418を200μg/mlを添加し継代した。作製した細胞株はpCMV26/Mock発現ベクターを導入したKK1細胞株(KK1/pCMV26-#7、KK1/pCMV26-#11)、NDRG2/pCMV26発現ベクターを導入したKK1細胞株(KK1/NDRG2-#5、KK1/NDRG2-#6)、pCMV26/Mock発現ベクターを導入したSu9T-01細胞株(Su9T-01/pCMV26-#2、Su9T-01/pCMV26-#4)、pCMV26/NDRG2発現ベクターを導入したSu9T-01細胞株(Su9T-01/NDRG2-#5、Su9T-01/NDRG2-#6)である。
ATL細胞はFlower様核を有する特異的な性質を持っている。上記のようにして得られたNDRG2安定発現ATL細胞を、まず細胞内局在の検討に供した。核はDAPI染色(青)をNDRG2はNDRG2をウサギ抗ヒトNDRG2抗体と二次抗体としてフルオロセン蛍光標識ヤギ抗ウサギIgG抗体を用いて染色(緑)した。
樹立したATL細胞株を用いて、各細胞株(1 x 105 個/ml)をそれぞれの継代に使用する培養液中で培養し、24 時間間隔でトリパンブルー染色法により生細胞数を算出した。また2 日間隔で10分の1倍希釈継代にすることで2週間培養した。細胞は37 度、5% 二酸化炭素下で静置培養した。
生体内での白血病細胞の増殖能を検討するために、Su9T-01/pCMV26-#4細胞株 (1 x 107 個)及び Su9T-01/NDRG2-#6細胞株 (1 x 107 個)を免疫不全マウス(NOD/Shi-scid, IL-2Rγnull (NOG)) 17)の経静脈及び腹腔投与を行なった。マウスが死亡後病理解剖し、病理学的、組織免疫染色、生化学的解析を行なった。
NDRG2とATL細胞の増殖や悪性化との関連性をさらに検証し、その治療効果を確認するために、NDRG2導入細胞株を用いて、インビトロ細胞増殖能とインビボマウス移植による腫瘍死による生存機関の比較を行なった。KK1-NDRG2安定発現株(KK1/NDRG2-#5、KK1/NDRG2-#6)はコントロール細胞(KK1、KK1/pCMV26-#7、KK1/pCMV26-#11)と比較しインビトロ増殖能が有意に低下していた。Su9T-01細胞株において同様にSu9T-01-NDRG2安定発現株(Su9T-01/NDRG2-#5、Su9T-01/NDRG2-#6)はコントロール細胞(SuT-01、Su9T-01/pCMV26-#2、Su9T-01/pCMV26-#4)と比較し同様に増殖能が低下している事が分かった(図10a)。そこで、さらに、Su9T-01-NDRG2安定発現株(Su9T-01/NDRG2-#6)及び、コントロール細胞(Su9T-01/pCMV26-#4)をNOD/Shi-scid, IL-2Rγnull (NOG) マウスそれぞれ6匹に移植し、経過観察を行なった。その結果、コントロールマウス群では、平均50%生存期間が53日日に対して、NDRG2安定発現株を移植したマウス群で、平均50%生存期間が84日と有意に延長しており(p<0.02)、さらに3匹は200日以上の長期生存を果たしている(図10b)。以上の結果から、NDRG2安定発現ATL細胞は、インビトロにおける細胞増殖能が低下し、さらにインビボにおいて白血病進展が遅延することがわかり、NDRG2の治療への応用可能性が確認できた。
Claims (4)
- 被験者のHTLV−1感染および/またはATL発症の有無の検出方法であって、
被験者由来の細胞中のNDRG2の発現レベルを測定し、コントロール細胞中のNDRG2の発現レベルと比較する工程;および
該比較において、被験者由来の細胞中のNDRG2の発現レベルが、コントロール細胞中のNDRG2の発現レベルより低下していることを指標として、被験者がHTLV−1の感染者および/またはATL発症者であると決定する工程
を含む、検出方法。 - 被験者のHTLV−1感染および/またはATL発症の有無の検出方法であって、
被験者由来の細胞中のNDRG2プロモーターのメチル化を測定する工程;およびメチル化がされていることを指標として、被験者がHTLV−1の感染者および/またはATL発症者であると決定する工程
を含む検出方法。 - NDRG2タンパク質またはそのペプチド断片に対する抗体を含むHTLV−1感染および/またはATL診断薬。
- NDRG2をコードするポリヌクレオチドが細胞内に導入され得る形態で含まれる、ATLの予防および/または治療のための薬剤。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2008031855A JP4751407B2 (ja) | 2008-02-13 | 2008-02-13 | ヒトt細胞白血病ウイルス感染あるいは成人t細胞白血病発症の検出法、その診断薬、ならびにその予防および治療の為の薬剤 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
JP2008031855A JP4751407B2 (ja) | 2008-02-13 | 2008-02-13 | ヒトt細胞白血病ウイルス感染あるいは成人t細胞白血病発症の検出法、その診断薬、ならびにその予防および治療の為の薬剤 |
Publications (2)
Publication Number | Publication Date |
---|---|
JP2009189271A JP2009189271A (ja) | 2009-08-27 |
JP4751407B2 true JP4751407B2 (ja) | 2011-08-17 |
Family
ID=41071929
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
JP2008031855A Expired - Fee Related JP4751407B2 (ja) | 2008-02-13 | 2008-02-13 | ヒトt細胞白血病ウイルス感染あるいは成人t細胞白血病発症の検出法、その診断薬、ならびにその予防および治療の為の薬剤 |
Country Status (1)
Country | Link |
---|---|
JP (1) | JP4751407B2 (ja) |
Families Citing this family (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
JP5492696B2 (ja) * | 2010-07-26 | 2014-05-14 | 大屋敷 一馬 | 一分子蛍光分析法によるdnaのメチル化度の決定方法 |
JP6051594B2 (ja) * | 2011-05-20 | 2016-12-27 | 国立大学法人 大分大学 | 糖質プロファイリングによる血液腫瘍診断のための診断基準を得る方法 |
JP6498114B2 (ja) * | 2013-04-25 | 2019-04-10 | 国立大学法人 宮崎大学 | 成人t細胞白血病の診断方法および成人t細胞白血病の治療薬のスクリーニング方法 |
-
2008
- 2008-02-13 JP JP2008031855A patent/JP4751407B2/ja not_active Expired - Fee Related
Also Published As
Publication number | Publication date |
---|---|
JP2009189271A (ja) | 2009-08-27 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
JP5901046B2 (ja) | OATP1B3mRNAの新規な選択的スプライシングバリアント | |
WO2014017491A1 (ja) | Cep55遺伝子とret遺伝子との融合遺伝子 | |
JP2001510987A (ja) | 哺乳動物のCD97αサブユニットを含む炎症および脈管形成を阻害するための方法および組成物 | |
AU2016248317A1 (en) | Methods for treating myeloproliferative disorders | |
AU2012273153A1 (en) | Compositions and methods for the therapy and diagnosis of cancer | |
AU2006241374A1 (en) | Novel Stra6 polypeptides | |
JP2022060484A (ja) | 悪性リンパ腫又は白血病の罹患の有無の判別方法並びに白血病の治療及び/又は予防のための薬剤 | |
US20210087247A1 (en) | Mps peptides and use thereof | |
JP3771218B2 (ja) | 細胞老化関連核酸配列及びタンパク質 | |
US20060134119A1 (en) | Drugs containing galectin 9 | |
JP4751407B2 (ja) | ヒトt細胞白血病ウイルス感染あるいは成人t細胞白血病発症の検出法、その診断薬、ならびにその予防および治療の為の薬剤 | |
WO2012015836A1 (en) | Highly potent peptides to control cancer and neurodegenerative diseases | |
EP2488548A1 (en) | A low density lipoprotein-related protein 1 splice variant as cancer marker | |
JPWO2005052156A1 (ja) | 肝癌の検出方法及び肝癌診断薬並びに癌治療薬 | |
JP4532273B2 (ja) | 癌に関係するタンパク質 | |
WO2011038689A1 (zh) | 人类新细胞因子VSTM1-v2及其应用 | |
JP5517078B2 (ja) | Ptenのリン酸化抑制剤又は脱リン酸化剤 | |
JP6498114B2 (ja) | 成人t細胞白血病の診断方法および成人t細胞白血病の治療薬のスクリーニング方法 | |
WO2018101950A1 (en) | Methods and compositions for the diagnosis of acute myeloid leukemia | |
JPWO2005021739A1 (ja) | Nox1ポリペプチドに対する抗体、Nox1遺伝子を利用したガン診断方法、及びガン増殖抑制物質のスクリーニング方法 | |
EP3969033A1 (en) | Mps modified peptides and use thereof | |
De Benedetti et al. | A Heterozygous | |
US20030077603A1 (en) | Methods and compositions for diagnosis of hepatoma | |
EP1661990A1 (en) | Polypeptide specific to liver cancer, polynucleotide encoding the polypeptide and rna molecule inhibiting the expression of the polypeptide | |
WO2005019257A1 (en) | A protein involved in pancreatic cancer |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20110210 |
|
A621 | Written request for application examination |
Free format text: JAPANESE INTERMEDIATE CODE: A621 Effective date: 20110210 |
|
A871 | Explanation of circumstances concerning accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A871 Effective date: 20110210 |
|
A975 | Report on accelerated examination |
Free format text: JAPANESE INTERMEDIATE CODE: A971005 Effective date: 20110308 |
|
A131 | Notification of reasons for refusal |
Free format text: JAPANESE INTERMEDIATE CODE: A131 Effective date: 20110325 |
|
A521 | Request for written amendment filed |
Free format text: JAPANESE INTERMEDIATE CODE: A523 Effective date: 20110407 |
|
TRDD | Decision of grant or rejection written | ||
A01 | Written decision to grant a patent or to grant a registration (utility model) |
Free format text: JAPANESE INTERMEDIATE CODE: A01 Effective date: 20110426 |
|
A61 | First payment of annual fees (during grant procedure) |
Free format text: JAPANESE INTERMEDIATE CODE: A61 Effective date: 20110520 |
|
R150 | Certificate of patent or registration of utility model |
Free format text: JAPANESE INTERMEDIATE CODE: R150 Ref document number: 4751407 Country of ref document: JP Free format text: JAPANESE INTERMEDIATE CODE: R150 |
|
FPAY | Renewal fee payment (event date is renewal date of database) |
Free format text: PAYMENT UNTIL: 20140527 Year of fee payment: 3 |
|
S533 | Written request for registration of change of name |
Free format text: JAPANESE INTERMEDIATE CODE: R313533 |
|
R360 | Written notification for declining of transfer of rights |
Free format text: JAPANESE INTERMEDIATE CODE: R360 |
|
S533 | Written request for registration of change of name |
Free format text: JAPANESE INTERMEDIATE CODE: R313533 |
|
R370 | Written measure of declining of transfer procedure |
Free format text: JAPANESE INTERMEDIATE CODE: R370 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313117 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
S111 | Request for change of ownership or part of ownership |
Free format text: JAPANESE INTERMEDIATE CODE: R313117 |
|
R350 | Written notification of registration of transfer |
Free format text: JAPANESE INTERMEDIATE CODE: R350 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
R250 | Receipt of annual fees |
Free format text: JAPANESE INTERMEDIATE CODE: R250 |
|
LAPS | Cancellation because of no payment of annual fees |