図1：骨肉腫細胞株と正常細胞株におけるmiR-1発現量
A：骨肉腫細胞株（MG63、Saos2、G292）と正常細胞株（hFOB1.19）のmiR-1発 現量。B：骨肉腫細胞株（MG63）へのmiR-1（50nM）導入による48時間でのmiR-1 発現量の変化。*p<0.05。
図2：miR-1 導入後の細胞増殖能および細胞周期と細胞死の解析
骨肉腫細胞株（MG63）へのmiR-1（50nM）・NC-miRNA（50nM）導入による細 胞増殖能・細胞周期および死細胞の評価。A：24、48、72時間での細胞増殖能。B：
48時間での細胞周期の割合。C：qRT-PCRによるp21発現量解析。D：Western blot による48時間でのp21、p53、リン酸化p53、p73発現解析。*p<0.05、**p<0.01。
図3：miR-1 導入後のPAX3発現解析
骨肉腫細胞株（MG63）へのmiR-1（50nM）・NC-miRNA（50nM）導入によるPAX3 への影響。A：PAX3におけるmiR-1配列並びに標的領域。太線はPAX3遺伝子の 3’-UTR（1521bp）を意味する。B（qRT-PCR）、C（Western blot）：48時間でのPAX3 発現解析。
図4：siPAX3導入後の細胞増殖能および細胞周期と細胞死の解析
骨肉腫細胞株（MG63）へのsiPAX3（10nM）・NC-siRNA（10nM）導入による細 胞増殖能・細胞周期および死細胞の評価。A（qRT-PCR）、B（Western blot）：48 時間でのPAX3発現解析。C：24、48、72時間での細胞増殖能。D：48時間での細 胞周期の割合。
E（qRT-PCR）、F（Western blot）：p21発現量解析。*p<0.05、**p<0.01。
図5：in vivoにおけるmiR-1の抗腫瘍効果と増殖活性の解析
マウス皮下腫瘍モデル（骨肉腫細胞株：MG63）作成し、miR-1（25nM）または
NC-miRNA（25nM）をそれぞれ腫瘍周囲に投与（各n=6）。miRNAの投与は0、7、
14日の計3回行った。A：腫瘍体積の経時的変化。B：腫瘍組織のKi67染色。C：
スコアリングによる増殖活性の評価。*p<0.05、**p<0.01。
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図6：miR-1導入後の遊走・浸潤能の解析
骨肉腫細胞株（MG63）へのmiR-1（50nM）・NC-miRNA（50nM）導入による遊 走・浸潤能の評価。A、B：Wound healing assay による24時間、48時間での遊走率 の解析。C、D：Matrigel invasion assayによる48時間後の浸潤細胞数の解析（▲：
浸潤骨肉腫細胞）。**p<0.01。
図7：miR-1導入後のSlug発現解析
骨肉腫細胞株（MG63）へのmiR-1（50nM）・NC-miRNA（50nM）導入によるSlug への影響。A：SlugにおけるmiR-1配列並びに標的領域。太線はSlug遺伝子の3’-UTR
（1127bp）を意味する。B（qRT-PCR）、C（Western blot）：48時間でのSlug発現 解析。**p<0.01。
図8：siSlug導入による遊走・浸潤能の解析
骨肉腫細胞株（MG63）へのsiSlug（30nM）・NC-siRNA（30nM）導入による遊 走・浸潤能の評価。A（qRT-PCR）、B（Western blot）：48時間でのSlug発現量解 析。C、D：Wound healing assay による24時間、48時間での遊走率の解析。E、F： Matrigel invasion assayによる48時間後の浸潤細胞数の解析（▲：浸潤骨肉腫細胞）。
G（qRT-PCR）、H（Western blot）：48時間でのEcad、Ncad、OBcad発現解析。*p<0.05、
**p<0.01。
図9：siOBcad導入による遊走・浸潤能の解析
骨肉腫細胞株（MG63）へのsiOBcad（50nM）・NC-siRNA（50nM）導入による 遊走・浸潤能の評価。A（qRT-PCR）、B（Western blot）：48時間でのOBcad発現 解析。C、D：Wound healing assay による24時間、48時間での遊走率の解析。E、F：
Matrigel invasion assayによる48時間後の浸潤細胞数の解析（▲：浸潤骨肉腫細胞）。
**p<0.01。
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13. 研究業績
藤井 亮太 I. 発表
① 一般発表 4
② 特別発表 0 II. 論文
① 原著論文 1（単0 ／共1 ）
② 症例報告 1（単0 ／共1 ）
③ 総説 なし
III. 著書 なし