アッセイID: U130 | 報告日: 2009/11/05 | 所属: 国立がんセンター研究所 | アッセイ者名: 河野隆志 | アッセイ名: in vivo DNA2本鎖切断修復(NHEJ)アッセイ |
概要 ヒト肺がん細胞株H1299にI-SceI制限酵素の切断基質DNAを安定導入したクローンを用い、I-SceI制限酵素一過性発現による非相同末端結合修復活性の測定を行った。このI-SceI制限酵素発現の際に、阻害剤を添加し、非相同末端結合修復活性の変動を測定した。現時点で、数個の阻害剤が修復反応を抑えるという結果が得られている。この結果は、複数の蛋白質が非相同末端結合に関わる可能性を示しており、今後の非相同末端結合修復の分子機構の解明、新規がん治療法の開発に示唆を与える結果となった。しかしながら、同阻害剤群は細胞のvaibilityへの影響が大きいことから、修復への特異性を更に検討する必要があると考えられた。 |
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評価のクライテリア 独自に開発したアッセイ系を用い、修復により翻訳されるEGFPタンパク質の発現細胞のfractionの減少を指標とした。 |
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発表論文 Histone acetylation by CBP and p300 at double-strand break sites facilitates SWI/SNF chromatin remodeling and the recruitment of non-homologous end joining factors. | レポート提出者によるレポート内容の最終確認日: 2013/10/28 |
Well | KIT 1 | KIT 2 | KIT 3 | |||
compound | 結果 | compound | 結果 | compound | 結果 | |
1 - A | none (DMSO) | none (DMSO) | none (DMSO) | |||
1 - B | 5-FU | Pifithrin-a (cyclic) | ABT-702 | |||
1 - C | Bestatin Hydrochloride | PRIMA-1 | Akt Inhibitor IV | |||
1 - D | Bleomycin sulfate | Finasteride | Akt Inhibitor VIII, Isozyme-Selective, Akti-1/2 | |||
1 - E | Cisplatin | Aminoglutethimide | Akt Inhibitor XI | |||
1 - F | Methotrexate | Formestane | compound C | |||
1 - G | Mitomycin C | Mifepristone | ATM/ATR kinase inhibitor | |||
1 - H | Vinblastine sulfate | TOFA | ATM kinase inhibitor | |||
2 - A | Paclitaxel | Amastatin | Aurora kinase/cdk inhibitor | |||
2 - B | Flutamide | Actinonin | Aurora kinase inhibitor II | |||
2 - C | Daunorubicin, HCl | Oligomycin | Aurora kinase inhibitor III | |||
2 - D | Doxorubicin, HCl | Bafilomycin A1 | AG957 | |||
2 - E | Tamoxifen, citrate | HA 14-1 | LFM-A13 | |||
2 - F | Actinomycin D | BH3I-1 | Terreic acid | |||
2 - G | Camptothecin | LFM-A13 | KN-93 | |||
2 - H | Aclarubicin | Terreic acid | KN-62 | |||
3 - A | Etoposide (VP-16) | E-64d | Lavendustin C | |||
3 - B | Cytochalasin D | ALLN | Kenpaullone | |||
3 - C | 2',5'-dideoxyadenosine | CA-074 | purvalanol A | |||
3 - D | AKT inhibitor | Pepstatin A | Olomoucine | |||
3 - E | NL-71-101 | Z-GLF-CMK | Alsterpaullone, 2-cyanoethyl | |||
3 - F | AG957 | RS 102895 | Cdk1/2 inhibitor III | |||
3 - G | KN93 | SB 328437 | Cdk2/9 inhibitor | |||
3 - H | Z-VAD-FMK | SB 225002 | NU6102 | |||
4 - A | Kenpaullone | AMD3100 octahydrochloride | Cdk4 inhibitor | |||
4 - B | Purvalanol A | NSC95397 | NSC625987 | |||
4 - C | 3-ATA | SC-ααδ9 | SB218078 | |||
4 - D | Olomoucine | Amiloride | isogranulatimide | |||
4 - E | TBB | Lidocaine | Chk2 inhibitor | |||
4 - F | Sulindac sulfide | Monensin | Chk2 inhibitor II | |||
4 - G | Valeryl salicylate | Ouabain | Ellagic acid (Dihydrate) | |||
4 - H | NS-398 | Sanguinarine | TBB | |||
5 - A | Sodium salicylate | Glibenclamide | DMAT | |||
5 - B | Theophylline | Dequalinium | D4476 | |||
5 - C | Azacytidine | Diazoxide | TG003 | |||
5 - D | Aphidicolin | Valinomycin | Diacylglycerol kinase inhibitor II | |||
5 - E | AG1478 | Nigericin | IC60211 | |||
5 - F | Genistein | Diltiazem | TX-1918 | |||
5 - G | Manumycin A | Nifedipine | BPIQ-Ⅱ | |||
5 - H | FTI-276 | Verapamil | AG1478 | |||
6 - A | SU1498 | PGP-4008 | AG490 | |||
6 - B | GGTI-286 | Fumitremorgin C | SU4984 | |||
6 - C | Dexamethasone | A23187 | SU5402 | |||
6 - D | GSK-3 inhibitor II | Ionomycin | Flt-3 Inhibitor | |||
6 - E | Scriptaid | Thapsigargin | cFMS Receptor Tyrosine Kinase Inhibitor | |||
6 - F | Trichostatin A | t-Butylhydroquinone (BHQ) | SU6656 | |||
6 - G | AG825 | N-phenylanthranilic acid | GSK-3 inhibitor IX | |||
6 - H | Cycloheximide | DIDS | 1-Azakenpaullone | |||
7 - A | Lovastatin | SB 218078 | Indirubin-3'-monoxime | |||
7 - B | Radicicol | Debromohymenialdisine (DBH) | AG825 | |||
7 - C | 17-AAG | Rotenone | AG1024 | |||
7 - D | AG1024 | Antimycin A1 | AGL 2263 | |||
7 - E | 1400W, HCl | Leptomycin B* | BMS-345541 | |||
7 - F | AMT, HCl | R59022 | IKK-2 inhibitor VI | |||
7 - G | AG490 | Dioctanoylglycol | IRAK-1/4 inhibitor | |||
7 - H | Cucurbitacin I | RHC80267 | JAK Inhibitor I | |||
8 - A | SP600125 | Xanthohumol | JAK3 Inhibitor VI | |||
8 - B | Damnacanthal | C75 | SP600125 | |||
8 - C | PD 98059 | Cerulenin | JNK inhibitor VIII | |||
8 - D | U0126 | Tunicamycin | Damnacanthal | |||
8 - E | Fumagillin | Deoxynojirimycin | PP2 | |||
8 - F | GM 6001 | Swainsonine | ERK inhibitor II | |||
8 - G | N-Acetyl-L-cysteine | LY 83583 | PD98059 | |||
8 - H | Aminoguanidine, HCl | ODQ | U-0126 | |||
9 - A | L-NMMA | Anacardic acid | MEK inhibitor I | |||
9 - B | PD169316 | Chetomin | SU11274 | |||
9 - C | SB 203580 | Dimethyloxalylglycine | ML-7 | |||
9 - D | Rapamycin | HR22C16 | SB202190 | |||
9 - E | NU1025 | Monastrol | SB239063 | |||
9 - F | Benzamide | Nordihydroguaiaretic acid (NDGA) | AG1296 | |||
9 - G | D609 | ETYA | SU11652 | |||
9 - H | IBMX | Baicalein | PDGF receptor tyrosine kinase inhibitor V | |||
10 - A | Ro-20-1724 | Nutlin-3 | PDGF receptor tyrosine kinase inhibitor IV | |||
10 - B | Zaprinast | MDM2 inhibitor | LY-294002 | |||
10 - C | AG1296 | Phenelzine | Wortmannin | |||
10 - D | LY294002 | Deprenyl | H-89 | |||
10 - E | Wortmannin | Decylubiquinone | 4-cyano-3-methylisoquinoline | |||
10 - F | H-89, HCl | Ro 5-4864 | Bisindolymaleimide I, HCl | |||
10 - G | Bisindolymaleimide I, HCl | Lonidamine | Go7874 | |||
10 - H | H-7 | ML-7 | Rp-8-CPT-cGMPS | |||
11 - A | Staurosporine | Benzylguanine | KT5823 | |||
11 - B | cPLA2inhibitor | DFMO | PKR inhibitor | |||
11 - C | OBAA | KT 5823 | RAF1 kinase inhibitor I | |||
11 - D | Cantharidin | Rp-8-CPT-cGMPS | ZM 336372 | |||
11 - E | Cytostatin | MK 886 | H-1152 | |||
11 - F | Cyclosporin A | Clofibrate | Y-27632 | |||
11 - G | FK-506 | BADGE | radicicol | |||
11 - H | MG-132 | Troglitazone | PP1 analog | |||
12 - A | Lactacystin | AZT | Syk inhibitor | |||
12 - B | Hydroxyurea | Nalidixic acid | SB431542 | |||
12 - C | HA1077 | a-Amanitin | TGF-b RI kinase inhibitor II | |||
12 - D | Y27632 | MST-312 | Tpl2 kinase inhibitor | |||
12 - E | PP1 (analog) | b-Rubromycin | TrkA inhibitor | |||
12 - F | PP-H | SB 431542 | VEGFR receptor tyrosine kinase inhibitor II | |||
12 - G | Nocodazole | N1,N12-Diethylspermine (BESpm) | VEGF recptor 2 kinase inhibitor I | |||
12 - H | Dephostatin | Fumonisin B1 | SU1498 |