The mechanism by which these Midostaurin datasheet miRNA-associated SNPs affect HBV-related hepatocarcinogenesis should be further studied. Disclosures: Kazuaki Chayama – Consulting: Abbvie; Grant/Research Support: Dainippon Sumitomo, Chugai, Mitsubishi Tanabe, DAIICHI SANKYO, Toray, BMS, MSD; Speaking and Teaching: Chugai, Mitsubishi Tanabe, DAIICHI SANKYO, KYO-RIN, Nihon Medi-Physics, BMS, Dainippon Sumitomo, MSD, ASKA, Astellas, AstraZeneca, Eisai,

Olympus, GlaxoSmithKline, ZERIA, Bayer, Minophagen, JANSSEN, JIMRO, TSUMURA, Otsuka, Taiho, Nippon Kayaku, Nippon Shin-yaku, Takeda, AJINOMOTO, Meiji Seika, Toray The following people have nothing to disclose: Daiki Miki, Hidenori Ochi, C. Nelson Hayes, Hiromi Abe, Tomokazu Kawaoka, Atsushi Ono, Sakura Akamatsu, Takashi Nakahara, Noriaki Seki, Eisuke Murakami, Yizhou Zhang, Takuro Uchida, Yohji Honda, Keiichi Masaki, Hiromi Kan, Masataka Tsuge, Nobuhiko Hiraga, Michio Imamura, Yoshiiku Kawakami, Hiroshi Aikata, Michiaki Kubo Background/Aims: Cyclin D1 plays an important role in the integration of mitogenic signals and promotion restriction point progression during G1 phase. Amplification and overexpression of cyclin D1 occur in tumorigenesis of several types of human cancers, including hepatocellular carcinoma (HCC). Our previous studies have shown that the intrabody against

cyclin D1 (Intra-AD1) can target cyclin D1 and inhibit the growth and proliferation of HCC. The present study is designed isometheptene to examine the underlying Inhibitor Library solubility dmso molecular mechanisms. Methods: A single-chain fragment of antibody variable region (scFv) against cyclin D1 (AD1) was prepared by phage display technique. Subsequently, an expression plasmid pIntra-AD1

habouring an endoplasmic reticulum (ER)-retained scFv gene against human cyclin D1 (Intra-AD1) was constructed. The human AD1 and cyclin D1 were expressed and prepared by affinity purification from the E.coli. The mimic epitope peptides recognized by AD1 were obtained by phage peptides library display technique. To verify the results, the spatial structure of AD1 was modeled and docked with cyclin D1 (from the PDB database) by computer software. Co-immunoprecipitation analysis was used to investigate whether the AD1 affected the interaction between cyclin D1 and CDK4 or pRB. The mRNA level was detected by Q-PCR. Results: Truncated mutation assay showed that the epitopes recognized by anti-cyclin D1 scFv (AD1) were in its N-terminal before amino acid A120. Results from phage peptides library display and sequence alignment showed that the epitopes on cyclin D1 was in its N-terminal including the pRB and CDK4 binding motifs. And this result was verified by computer modeling and docking. Moreover the key amino acids recognized by AD1 were N24, K33, K112, M113, K114, E115.