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國立成功大學「邁向頂尖大學計畫」
延攬優秀人才工作報告表
NCKU’s “Aim for the Top University Project”
Work Report Form for Distinguished Scholars
□續聘
continuation of employment ■離職
resignation
100 年 7 月 13 日更新
受聘者姓名
Name of the Employee
羅芳宜
Male Female聘 期
Period of EmploymentFrom100 年(y)08 月(m)01 日(d)
to 102 年(y)12 月(m)31 日(d)
研究或教學或科技研發與
管理計畫名稱
The project title of research,teaching, technology development and management
國家型生技醫藥科技計畫：
以固態腫瘤細胞模式及表基因
體技術探討新穎組蛋白去乙醯
酵素抑制劑及其接合癌症標靶
增效載體抑制腫瘤生長與轉移
之研究
計畫主持人
（申請單位主管）
Project Investigator (Head of Department/Center)計畫主持人：王憶卿
單位主管：呂增宏
補助延聘編號
Grant Number
HUA101-3-7-473
一、
研究、教學、科技研發與管理工作全程經過概述。
（由受聘人填寫）
Please summarize the entire research, teaching, or science and technology R&D and management work process (To be
completed by the employee)
Background: Overexpression and/or increased activity of histone deacetylases (HDACs) in
hematologic and solid malignancies make HDACs a potential therapeutic target for cancer treatment.
However, many HDAC inhibitors show serious adverse events in the course of treatment for solid
tumors.
Purpose: To evaluate the antitumor effects and toxicity of novel HDAC inhibitors, several candidate
compounds were examined by in vitro and in vivo models of human lung cancer.
Methodology/Results: HDAC inhibition was examined by in vitro HDAC activity assay and western
blot of acetylated histone and non-histone proteins. The cytotoxicity effect was examined in five human
lung cancer cell lines. The antiproliferatative mechanisms were investigated by flow cytometric cell
cycle analysis, apoptosis assay, and repair gene expression assay in cells treated with HDAC inhibitor
along or in combination with cisplatin. Mice with A549 tumor xenograft were used to evaluate effects
on tumor growth and tumor metastasis. Over 500 candidate compounds were screened by in vitro
HDAC activity assay to search for potent HDAC inhibitors. We found that novel HDAC inhibitor,
WJ-6K showed HDAC activity inhibition ability better or comparable to FDA-approved HDAC
inhibitor SAHA and led to increased HDAC target protein acetylation in lung cancer cells (Fig 1).
WJ-6K showed significant cytotoxicity in various lung cancer cell lines without affecting normal lung
cell IMR90 at the dose range examined (Fig 2). WJ-6K led to cancer cell death through induction of
cell cycle G2/M arrest and mitochondria-mediated apoptosis. We further confirmed that WJ-6K was
more effective than SAHA in inhibition of tumor growth in A549 xenograft animal model without
significant side effects (Fig 3). Combined treatment of WJ-6K and conventional chemotherapy agent,
cisplatin showed synergistic cytotoxicity in lung cancer cells including the cisplatin-resistant cell line
H1435 through impairment of DNA damage repair (Fig 4). Using trans-well invasion assays, we found
that WJ-6K inhibited cell invasion in A549 at non-cytotoxic doses. We further demonstrated that
WJ-6K pre-treatment significantly suppressed A549 metastases to lungs by experimental metastasis
assay in mice (Fig 5).
Conclusions: Our findings suggest that WJ-6K is a novel potent HDAC inhibitor which suppresses
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Figure 1. WJ-6K is a potent pan-HDAC inhibitor in vitro and in cells.
(A) HDAC activity is measured
by in vitro HDAC activity assay. Left: Total HDAC activity; Right: IC50 (nM) to individual
HDAC isoform after incubation of indicated concentrations of SAHA or WJ-6K. P＜0.05 is shown as *; P＜0.01 is shown as
**; P＜0.001 is shown as ***. (B) Western blot analysis showed
that WJ-6K treatment (2 hours) stimulated HDAC target proteins acetylation in a dose dependent manner in A549 and CL1-5 cells.
Figure 2. WJ-6K induces cancer-specific cell death.
WJ-6K effectively inhibited cell growth of various lung cancer cell lines without affecting the normal lung cell line IMR90. Points: mean: bars: ±SEM (n≧3).
Figure 3. WJ-6K suppresses tumor growth in animal model.
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Figure 4. WJ-6K shows a synergistic anti-tumor growth effect when combined with cisplatin in vivo.
Balb/c nude mice bearing the established A549 tumors were treated with 25mg/kg WJ-6K or 2mg/kg cisplatin alone or combined through intraperitoneal injection for three weeks. Left panel: The tumor sizes of mice were measured; Right panel: The measured tumor weight; Points, mean; bars, ±SEM (n=6). P＜0.05 is shown as *; P＜0.01 is shown as **.
Figure 5. WJ-6K suppresses cancer metastasis in vivo.
A549 cells were pre-treated with 1μM of SAHA or WJ-6K for 48 hours. After pre-treatment, A549 cells were injected intravenously via tail vein into NOD/SCID mice and observed for 60 days after cancer cell injection. The ability to lung metastases of A549 was suppressed after WJ-6K treatment. (A) Less A549 tumor nodules were found in the lungs of tested animals in WJ-6K treated group than in DMSO or SAHA treated group.(B) Representative H&E stain of lung tissues of
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二、
研究或教學或科技研發與管理成效評估（
由計畫主持人或單位主管填寫
）
Please evaluate the performance of research, teaching or science and technology R&D and management Work: (To be completed
by Project Investigator or Head of Department/Center)
(1)是否達到延攬預期目標？
Has the expected goal of recruitment been achieved?
Yes.
(2)研究或教學或科技研發與管理的方法、專業知識及進度如何？
What are the methods, professional knowledge, and progress of the research, teaching, or R&D and management work?
The applicant (Dr. Fang-Yi Lo) have conduct the cell culture, compound treatment,
cytotoxicity test, cell motility assay, and Western blot for acetylated histones and non-histone
proteins using one potential histone deacetylase inhibitor (HDACi), WJ-6K, in lung cancer cell
model. In addition, tumor growth inhibition of this HDACi is completed for A549 xenograft
assay.
(3)受延攬人之研究或教學或科技研發與管理成果對該計畫(或貴單位)助益如何？
How have the research, teaching, or R&D and management results of the employed person given benefit to the project (or your unit)?
The applicant (Dr. Fang-Yi Lo)’s studies on HDACi in the passed 5 months benefit a lot to the
project. The data of cell and animal models indicate the anti-cancer growth potential of the
current HDACi, WJ-6K. Studies on anti-metastasis using cell and animal models will be
further conduct with the help of Dr. Lo.
(4)受延攬人於補助期間對貴單位或國內相關學術科技領域助益如何？
How has the employed person, during his or her term of employment, benefited your unit or the relevant domestic academic field?
HDAC inhibitors have shown anticancer activities in many preclinical and clinical
investigations of human cancers. However, many HDAC inhibitors have serious adverse
events in the course of treatment for solid tumors. The applicant (Dr. Fang-Yi Lo)’s studies
on HDACi in the passed 5 months benefit a lot to approve the field of drug development.
(5)具體工作績效或研究或教學或科技研發與管理成果：
Please describe the specific work performance, or the results of research, teaching, or R&D and management work:
