Potential of new anti-cancer agents targeting the nuclear translocation signaling of HB-EGF C-terminal fragments

针对 HB-EGF C 端片段核转位信号的新型抗癌药物的潜力

• 批准号: 22590704
• 负责人: TANIDA Satoshi
• 金额: $ 3万
• 依托单位: Nagoya City University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2010
• 资助国家: 日本
• 起止时间: 2010 至 2012
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-22590704/
• 关键词: EGFファミリー; HB-EGF核移行シグナル; PLZF; Alphascreen assay; IL-8; HB-EGF-CTF; colitic cancer; 潰瘍性大腸炎; 炎症性腸疾患

Current treatment target toward advanced colorectal cancers is mainly focused on the epidermal growth factor receptor (EGFR) signaling, but its additive effects with chemotherapy are still limited. A disintegrin and metalloproteinase (ADAM) cleaves the proheparin-binding epidermal growth factor like growth factor (proHB-EGF). And soluble HB-EGF activates EGFR. In parallel, the carboxy-terminal fragment of proHB-EGF (HB-EGF-CTF) translocate into the inner nuclear membrane, subsequently exerts on the regulation of cell proliferation by binding nuclear promyelocytic leukemia zinc finger (PLZF) protein, a transcriptional repressor, thereby causing its nuclear export. We hypothesized that the inhibition of HB-EGF-CTF nuclear translocation may be a new strategy in preventing cell proliferation.Methods:12-O-tetradecanoylphorbor-13-acetate (TPA) was treated to activate ADAM. Nine-thousand chemical compounds were screened for their efficacies in blocking the binding of HB-EGF-CTF to promyelocyt … More ic leukemia zinc finger (PLZF) with Alphascreen system. The obtained candidates were then used to block the binding of HB-EGF-CTF to PLZF in colon cancer cells, HT29 and HCT116. Cell proliferation was investigated with a growth curve assay. The intracellular localization, and association between HB-EGF-CTF and PLZF, was assessed with immunofluorescent staining, and immunoprecipitation and Western blotting, respectively. The effects of obtained candidates on EGFR phosphorylation and on nuclear translocation of HB-EGF-CTF and export of PLZF during the angiotensin II type1 receptor (AT1R) knockdown were also investigated. Results: Telmisartan and candesartan were found to be potential candidates. Telmisartan inhibited TPA-induced cell proliferation stronger than candesartan. Telmisartan, but not candesartan blocked the nuclear translocation of HB-EGF-CTF, and binding of HB-EGF-CTF to PLZF, during TPA stimulation. Both telmisartan and candesartan did not inhibit TPA-induced EGFR phosphorylation, and telmisartan, but not candesartan, inhibited TPA-induced nuclear translocation of HB-EGF-CTF after knockdown of AT1R. Conclusions: The inhibition of HB-EGF-CTF nuclear translocation with telmisartan may be a novel strategy in preventing cell proliferation. Less

目前针对晚期结直肠癌的治疗靶点主要集中在表皮生长因子受体（EGFR）信号转导上，但其与化疗的叠加作用仍然有限。解整合素和金属蛋白酶（ADAM）切割前肝素结合表皮生长因子样生长因子（proHB-EGF）。可溶性HB-EGF激活EGFR。同时，proHB-EGF的羧基端片段（HB-EGF-CTF）转位到核内膜，通过与转录抑制因子--核早幼粒细胞白血病锌指蛋白（PLZF）结合，从而调控细胞增殖，导致其向核输出。方法：用12-O-十四烷酰基-13-乙酸磷酯（TPA）激活ADAM，用MTT法检测细胞增殖。筛选了9000种化合物，以确定其阻断HB-EGF-CTF与早幼粒细胞结合的有效性。 ...更多信息 用Alphascreen系统检测白血病锌指（PLZF）。然后将获得的候选物用于阻断结肠癌细胞HT 29和HCT 116中HB-EGF-CTF与PLZF的结合。用生长曲线测定法研究细胞增殖。细胞内定位，HB-EGF-CTF和PLZF之间的关联，分别用免疫荧光染色，免疫沉淀和Western印迹进行评估。还研究了获得的候选物对EGFR磷酸化和对HB-EGF-CTF的核转位以及在血管紧张素II 1型受体（AT 1 R）敲低期间PLZF的输出的影响。结果：替米沙坦和坎地沙坦是潜在的候选药物。替米沙坦对TPA诱导的细胞增殖的抑制作用强于坎地沙坦。在TPA刺激期间，替米沙坦（而非坎地沙坦）阻断HB-EGF-CTF的核转位以及HB-EGF-CTF与PLZF的结合。替米沙坦和坎地沙坦均不抑制TPA诱导的EGFR磷酸化，并且在AT 1 R敲低后，替米沙坦而非坎地沙坦抑制TPA诱导的HB-EGF-CTF核转位。结论：替米沙坦抑制HB-EGF-CTF核转位可能是抑制细胞增殖的新策略。少

项目成果

IBD 腸管炎症に関わる炎症性サイトカインによるEGFシグナルを介した大腸細胞増殖機序‐HB-EGF-C末端シグナルを標的とした網羅的薬剤探索‐

IBD 肠道炎症相关炎症细胞因子引起的 EGF 信号介导的结肠细胞增殖机制 - 针对 HB-EGF-C 末端信号的综合药物搜索 -

• 发表时间: 2011
• 作者: 谷田 諭史;溝下 勤 ;水島 隆史;城 卓志
• 通讯作者: 城 卓志

HB-EGF-C末端核移行シグナルをターゲットにした新規薬剤探索と細胞増殖抑制効果について.

寻找针对HB-EGF-C末端核转位信号的新药及其细胞增殖抑制作用。

• 发表时间: 2010
• 作者: 尾関啓司;谷田諭史;福田信治;東山繁樹;城 卓志
• 通讯作者: 城 卓志

HB-EGF-C末端核移行シグナルをターゲットにした新規薬剤探索と細胞増殖抑制効果について

寻找针对HB-EGF-C末端核转位信号和细胞增殖抑制作用的新药

• 发表时间: 2010
• 作者: Mikami Y;Kanai T;Sujino T;Ono Y,Hayashi A;Okazawa A;Kamada N,Matsuoka K;Hisamatsu T;Okamoto S,Takaishi H;Inoue N;Ogata H;Hibi T.;尾関啓司
• 通讯作者: 尾関啓司

IBD 腸管炎症に関わる炎症性サイトカインによるEGF signalを介した大腸癌細胞増殖機序 -EGF-C末端signalを標的とした新規薬剤探索-

IBD 炎症细胞因子参与肠道炎症引起EGF信号介导的结直肠癌细胞增殖机制 - 寻找针对EGF-C末端信号的新药 -

• 发表时间: 2011
• 作者: 尾関啓司;谷田諭史;溝下 勤;塚本宏延;城 卓志
• 通讯作者: 城 卓志

TGFbeta induces shedding of proHB-EGF and EGFR transactivasion though ADAM activation.

TGFbeta 通过 ADAM 激活诱导 proHB-EGF 和 EGFR 反式激活的脱落。

• 发表时间: 2010
• 作者: 海老正秀;城 卓志;神谷武;片岡洋望;久保田英嗣;溝下 勤;志村貴也;村上賢治;平田慶和;東山繁樹.
• 通讯作者: 東山繁樹.