RNA (RNA interference) Fire A, Mello C 1998 Nature 2006 (C elegans) unc-22 unc RNA 742 dsrna RNA mrna RNase Dicer RNA sirna(small inteferin

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1 microrna RNA(non-coding RNA 23,000 RNA:20, RNA 1/3 RNA RNA 1800 mir-122, mir-200 let-7

2 RNA (RNA interference) Fire A, Mello C 1998 Nature 2006 (C elegans) unc-22 unc RNA 742 dsrna RNA mrna RNase Dicer RNA sirna(small intefering RNA) sirna RISC (RNA-induced silencing complex) mrna

3 mirna, sirna

4 sirna mirna mirna mirna 6-8 mirna (Target Scan

5 Dicer, Drosha mrna Merritt. WM, Sood AK, N Eng J Med 2008 Dicer mrna RNA

6 mirna mir-124a is required for hippocampal axogenesis and retinal cone survival through Lhx2 suppression. Sanuki R, Furukawa T Nat Neurosci 2011 mir-124a mirna 25-48% mir124a

7

8

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10 C mirna mir-122 HCV RNA mir-122 mir-122 HCV RNA SPC3649 mir mir-122 Therapeutic silencing of microrna-122 in patients with chronic hepatitis C virus infection HCV C SPC3649 : high dose (5mg/kg), low dose(1mg/kg) 2 12 Robert E, Henrik O Science 2010

11 HCV RBA mir-122 Silencing of mir-122 by SPC3649 in chimpanzees with chronic hepatitis C virus infection. (A) Analysis of HCV RNA levels in HCV-infected chimpanzees during the study. The HCV titers are shown as genomic equivalents (GE) for the high-dose animals (4x0513, blue triangles; 4x0514, magenta diamonds) and low-dose animals (4x0267, orange squares; 4x0358, red dots) in serum (GE/ml, solid lines) and liver (GE/ μg liver RNA, dashed lines). The placebo and active treatment periods are indicated below. (B) Northern blot analysis of RNA from chimpanzee liver biopsies using LNA-modified probes detecting free and sequestered mir-122 (upper panel) and SPC3649 (lower panel). The first two lanes are positive controls for free mir-122 and preformed mir-122:spc3649 heteroduplexes, respectively. (C) Detection of sequence variants in the mir-122 seed sites (boxed) by deep sequencing of the HCV 5 NCR from the high-dose animals at four time points: baseline, end of treatment, viral rebound, and end of the follow-up period. (D) The two mir-122 seed sites (boxed) in the HCV 5 NCR are conserved in all HCV genotypes and subtypes (see Supporting Online Material for details).

12 Treatment of HCV-infected chimpanzees with SPC3649 was well tolerated. (A) Plasma trough levels of SPC3649. (B and C) Alanine aminotransferase (ALT) levels (B) and creatinine levels (C) in HCV-infected chimpanzees during the study. (D to G) Photomicrographs of hematoxylin and eosin stained sections from biopsies of a normal chimpanzee liver (D) and animal 4x0513 at week 4 (E), week 19 (F), and week 25 (G), respectively. A SPC B ALT(GPT) C

13 EMT (epithelial mesenchymal transition) E-cadherin vimentin,fibronectin, ZEB1(E-cadherin,ZEB2

14 retrovirus mir-200c mir-200c expression induces an EMT phenotype in UMUC3 bladder cancer cell line. A, phasecontrast microscopy of UMUC3 cells transduced with a mir-200c containing retrovirus (UMUC3/200c) or with an empty, control, retroviral construct (UMUC3-E). Bar, 100 µm. B, measurement of in vitro cell migration by "woundhealing" assay. Representative pictures for same single spot. The experiment was done twice in triplicate experiments. Bar, 400 µm. C, quantification of mir-200c in empty virus transduced (UMUC3-E) and mir-200c-transduced (UMUC3/200c) cells. Levels of mir-200c in UMUC5 cells are plotted for comparison. D, measurement by real-time RT-PCR of the epithelial and mesenchymal markers E-cadherin, ZEB1, and ZEB2 in empty virus transduced cells and mir-200c-transduced UMUC3 cells. E, confocal microscopy analysis of the UMUC3 series costained for ZEB1 or ZEB2 (red pixels) and nuclear DNA (green pixels). Note the downregulation of nuclear ZEB1 and ZEB2 in UMUC3 clones expressing the mir-200c. F, measurement of ERRFI-1 and E-cadherin protein levels by Western blot. Actin reprobing served as internal (loading) control. Bottom, absorbance relative values expressed as ratios between actin (internal control) and ERRFI-1. Note the up-regulation of E- cadherin and down-regulation of ERRFI-1 protein levels on mir-200c expression.

15 mir200c ERRFI-1(ErbB receptor inhibitor-1) EGFR C225(Cetuximab) p-mapk mir-200c expression reverses EGFR resistance in UMUC3 cells. A, confocal microscopy analysis of the UMUC3 series costained for ERRFI-1 (red pixels) and EGFR (green pixels) and a DNA dye, showing the nucleus (blue pixels). Note the yellow pixels (left) as a result of red and green pixels colocalization. B, immunoblot of autophosphorylated EGFR, total EGFR, and total ERRFI-1 in cells transduced with mir-200c from the experiment above. In this experiment, actin served as the internal control. C, top, immunoblot of pmapk, total MAPK, and total EGFR of the UMUC3 series. Cells grown in 2% serumsupplemented MEM were left untreated or treated with increased concentrations of cetuximab (C225) for 3 h. Bottom, absorbance relative values expressed as ratios between EGFR (internal control) and pmapk. D, cell proliferation measurement of the UMUC3 series using radioactive thymidine incorporation. Each experiment was done in at least two different triplicates.

16 mir-200 mir200 Tublin( β-catenin ZEB1 ErbB receptor inhibitor-1 Cyclin D1 E-cadherin EGFR

17

熊本大学学術リポジトリ Kumamoto University Repositor Title 炎症性腸疾患に対する HSF1 及び HSP70 の保護的役割 Author(s) 田中, 健一郎 Citation Issue date Type URL Thesis or Di

熊本大学学術リポジトリ Kumamoto University Repositor Title 炎症性腸疾患に対する HSF1 及び HSP70 の保護的役割 Author(s) 田中, 健一郎 Citation Issue date Type URL Thesis or Di 熊本大学学術リポジトリ Kumamoto University Repositor Title 炎症性腸疾患に対する HSF1 及び HSP70 の保護的役割 Author(s) 田中, 健一郎 Citation Issue date 2008-03-25 Type URL Thesis or Dissertation http://hdl.handle.net/2298/9448 Right 1

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