Poster Sessions Room P(A3-1) Sep. 25 (Thu.) 17:00-17:45
P5-3
P-1078 AREG regulates Warburg effect in colorectal cancer through the binding of MLX to ChoRE in the AREG promoter
Signal transduction (3)
Sung Ouk Nam1,2, Fusanori Yotsumoto2,3, Kohei Miyata1,2, Satoshi Fukagawa1,2, Yasunobu Kanamori4, Masahide Kuroki2,3, Shingo Miyamoto1,2 (1Dept. Obstet. & Gynecol. Sch. of Med., Fukuoka Univ., 2Ctr. Advanced Mol. Med., Fukuoka Univ., 3Dept. Biochem. Sch. of Med., Fukuoka Univ., 4Dept. Obstet. & Gynecol. Yamaguti Red Cross Hosp.)
シグナル伝達 (3)
Chairperson: Yasuyuki Nakamura (Div. Cancer Biol., Natl. Cancer Ctr. Res. Inst.) 座長：中村 康之（国立がん研究セ・研・腫瘍生物） P-1075 TAX1BP1 modulates antigen-specific antibody response through NF-κB pathway Midori Suzuki 1 , Emi Ikebe 2 , Hidekatsu Iha 2 , Shigeru Yanagi 1 , Nobuko Matsushita 1 ( 1 Lab. of Mol. Biochem.,Tokyo Univ. of Pharmacy and Life Science, 2Dept. of Microbiology,Faculty of Med.,Oita Univ.) TAX1BP1はNF-κB経路を制御することによって抗原特異的抗体産生に機能 する 鈴木 みどり 1 、池辺 詠美 2 、伊波 英克 2 、柳 茂 1 、松下 暢子 1 （1 東京薬科大 学・生命科学・分子生化学、2 大分大学・医・微生物学講座） E-mail:
[email protected]
AREGはプロモーター領域ChoREのMLX結合を介して大腸癌腫瘍形成時の Warburg effectを制御する 南 星旭 1,2 、四元 房典 2,3 、宮田 康平 1,2 、深川 怜史 1,2 、金森 康展 4 、黒木 政 秀 2,3 、宮本 新吾 1,2 （1 福岡大・医・産婦人科、2 福岡大・先端分子医学研究セ ンター、3 福岡大・医・生化学、4 山口赤十字病院産婦人科） E-mail:
[email protected]
Nuclear factor (NF)-κB is a key player of inducible transcription in inflammation, innate immunity. NF-κB pathway is up-regulated in many chronic inflammatory diseases and a variety of B-cell lymphoma, and persistent activation of NF-κB pathway results in pathological inflammation. TAX1BP1 is a ubiqutin binding protein, interacts with ubiquitin editing enzyme A20, functions in termination of NF-κB activity through the tumor necrosis factor receptor (TNFR) and Toll-like receptor (TLR) pathways. A20 and TAX1BP1 target RIP1and TRAF6, inter mediator of NF-κB pathway, for inactivation by cleaving K63-linked polyubiquitin chains and promoting K48-linked polyubiquitination result in proteasomemediated degradation. Here we generated TAX1BP1 deficient chicken DT40 B-lymphoma cell line, we showed that TAX1BP1 deficiency activated NF-κB pathway and resulted in the acceleration of inflammation and the failure of antigen-specific antibody response. Keywords: NF-kB, B-cell lymphoma
Amphiregulin (AREG) is essential for the tumor formation of colorectal cancer (CRC). Therefore, the genes associated with the expression of AREG are possibly involved in the molecular mechanisms of CRC. To detect the genes associated with AREG expression, we performed cDNA microarray analysis between 2-dimensional culture (DC) and 3DC in CRC cell lines. Upregulated and downregulated genes in 3DC compared with 2DC were picked up. These genes were involved in glycolysis using pathway analysis. Increased concentration of glucose enhanced the expression of GLUT1 and AREG as well as the activity of the HIF-1. The transfection of siRNA for AREG decreased the glucose uptake and lactate production. Using luciferase assay, the promoter region of AREG was determined between -130 and -180 bp upstream of the transcription start site, which contained carbohydrate response element (ChoRE). We focused on Max-like protein X (MLX), which binds to carbohydrate response elements involved in glucose metabolism. ChIP analysis revealed MLX binding to the AREG promoter. These data suggest that AREG plays a pivotal role in the Warburg effect of CRC through the binding of MLX to ChoRE in the AREG promoter. Keyword: Amphiregulin
P-1076 The role of GEP oncogene, Gα12/13, in the progression of ovarian cancer
P-1079 The role of the Erc/mesothelin on signal transduction in Lipid raft
Hiroshi Yagi, Kenzo Sonoda, Kiyoko Kato (Dept. OBGY., Sch. Med., Kyushu Univ.) 卵巣癌の進展におけるGEP oncogene(Gα 12/13 )の役割 八木 裕史、園田 顕三、加藤 聖子（九州大・医・産婦人科）
G protein-coupled receptors (GPCRs) and their ligands have been implicated in the progression of human cancers. In GPCR signaling through heterotrimeric G proteins, Gα12/13, encoded by the GEP oncogene, is closely linked to tumor progression. We herein evaluated the role of GEP oncogene in human ovarian cancer progression.Gα12/13 was highly expressed in human ovarian cancer tissues and cell lines. Inhibition of Gα12/13 signaling by expression of RGS domain of PDZ-RhoGEF in human ovarian cancer cell lines blocked cell proliferation and colony formation in vitro. GPCRs involved in cell proliferation stimulate the activity of the transcriptional co-activator YAP, which is a critical component of the Hippo signaling pathway that controls organ size in mammals. Synthetic biology approaches using mutants of GPCR and G protein showed that the activation of Gα13 signaling pathway induced YAP activation in ovarian cancer cell lines. Increased expression of GEP oncogenes, Gα12/13, can result in enhanced signaling downstream of GPCRs, thereby influencing proliferation of ovarian cancer. Keyword: G protein P-1077 Up-regulation of the Maspin Gene Expression by Transforming Growth Factor-β (TGF-β) in Human Cervical Carcinoma Cells Ariyaphong Wongnoppavich 1 , Kongthawat Chairatvit 2 ( 1 Dept. of Biochemistry, Fac. of Medicine, CMU, 2 Dept. of Oral Biology, Fac. of Dentistry, MU) E-mail:
[email protected]
Danqing Zhang1, Toshiyuki Kobayashi1,2, Okio Hino1,2 (1 Dept. Pathology and Oncology, Juntendo Univ., Sch. Med., 2 Dept. Molecular Pathogenesis, Juntendo Univ., Grad. Sch. Med.) 脂質ラフトにおけるシグナル伝達に関わるErc/mesothelinの役割 小橋（張） 丹青 1 、小林 敏之 1,2 、樋野 興夫 1,2 （1 順天堂大学医学部 病理・ 腫瘍学、2 順天堂大学 院医 分子病理病態学） E-mail:
[email protected]
Mouse Erc (Expressed in renal carcinoma) is a homologue of human mesothelin gene. Erc/mesothelin (hereafter named as Erc) is a GPIanchored membrane protein expressed in normal mesothelium and induced in several species of human malignant tumors, as well as in renal tumors of Tsc2 KO mice. To investigate the role of Erc in signal pathway, Erc-deficient Tsc2 KO mouse renal tumor cell line was transfected with Erc expression vector or empty vector and then their lysates were analyzed by cell fractionation. Erc/mesothelin was found in lipid raft fraction. We also found that the phosphorylation of EGF receptor (EGFR) was reduced but total EGFR was remarkably increased in Ercdeficient cells compared with Erc-restored cells. Multiple bands were observed in Western blots of EGFR on lipid raft, the lowest band was clearly observed in Erc-restored cells. The expression level of Egfr mRNA in Erc-deficient cells was higher than in Erc-restored cells as revealed by RT-PCR, real time PCR and GeneChip exon array. It is plausible that the expression of EGFR may be regulated by feedback from Erc-regulated signal pathways other than mTOR axis involved in the Tsc2 mutationinduced tumorigenesis. Keywords: Erc gene, EGF receptor
Maspin or SERPIN B5 acts as a tumor suppressor via its inhibitory effect on cancer cell migration, invasion and metastasis. The role of maspin on tumorigenesis is likely to be tissue specific since its expression is paradoxically up-regulated in many malignant cells. The regulation of maspin expression might depend on the genetic/epigenetic background and specific microenvironment of carcinoma cells. In this study, we showed that transforming growth factor-β(TGF-β) induced maspin expression at both transcript and protein levels in human cervical carcinoma cells (HeLa). TGF-βactivated the Smad signaling pathway since the phosphorylated Smad2/3 was detected upon the cytokine treatment in HeLa. Pretreatment of several inhibitors showed that U0126 or SB202190 could block the effect of TGF-βon maspin expression in the carcinoma cells whereas PDTC or wartmannin did not. Interestingly, all tested inhibitors did not abolish the TGF-β-induced phosphorylation of Smad2/3. In conclusion, induction of maspin expression by TGF-βin human cervical carcinoma likely mediates through the non-Smad signaling pathway in which MEKK3/4 and p38 MAPK are required as intermediate signaling molecules. Keywords: Maspin, TGF-beta
272
