Browsing by Author "Liu, Kuang-Kai"

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7-Chloro-6-piperidin-1-yl-quinoline-5,8-dione (PT-262), a novel ROCK inhibitor blocks cytoskeleton function and cell migration
(2011-04-01)
Tsai, Chih-Chien
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Liu, Huei-Fang
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Hsu, Kai-Cheng
;
Yang, Jinn-Moon
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Chen, Chinpiao
The 5,8-quinolinediones are precursors for producing multiple types of bioactive products. In this study, we investigated a new compound derived from 5,8-quinolinediones, 7-chloro-6-piperidin-1-yl-quinoline-5,8-dione (designated as PT-262), which markedly induced cytoskeleton remodeling and migration inhibition in lung carcinoma cells. Comparison with various cytoskeleton inhibitors, including paclitaxel, colchicine and phallacidin, the cell morphology following treatment with PT-262 was similar to phallacidin on the cell elongation and abnormal actin polymerization. However, PT-262 did not directly bind to actin filaments. ROCK (Rho-associated coiled-coil forming protein kinase) is a downstream effector of RhoA to mediate the phosphorylation of myosin light chain (MLC) and cytoskeleton reorganization. The RhoA-ROCK-MLC pathway has been shown to promote cancer cell migration and metastasis. Interestingly. PT-262 was more effective on inhibiting ROCK kinase activities than specific ROCK inhibitors Y-27632 and H-1152. PT-262 induced cytoskeleton remodeling and migration inhibition in A549 lung carcinoma cells. The total MLC and phosphorylated MLC proteins and stress fibers were blocked after treatment with PT-262. Nonetheless, the RhoA protein and GTPase activity were not altered by PT-262. A computational model suggests that PT-262 interacts with the ATP-binding site of ROCK protein. Together, these findings demonstrate that PT-262 is a new ROCK inhibitor. (C) 2011 Elsevier Inc. All rights reserved.
A novel synthetic compound overcomes drug resistance on EGFR (T790M) and cancer stemness in human non-small cell lung cancer
(2017-04-01)
Pan, Pei Ying
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Chang, Chien-Jen
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Liu, Kuang-Kai
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Chan, Yu-Chi
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Chen, Yan-Ting
Alpha-bungarotoxin binding to target cell in a developing visual system by carboxylated nanodiamond
(2008-05-21)
Liu, Kuang-Kai
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Mei-Fang Chen
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Chen, Po-Yi
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Lee, Tony J. F.
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Cheng, Chia-Liang
Biological molecules conjugating with nanoparticles are valuable for applications including bio-imaging, bio-detection, and bio-sensing. Nanometer-sized diamond particles have excellent electronic and chemical properties for bio-conjugation. In this study, we manipulated the carboxyl group produced on the surface of nanodiamond (carboxylated nanodiamond, cND) for conjugating with alpha-bungarotoxin (alpha-BTX), a neurotoxin derived from Bungarus multicinctus with specific blockade of alpha7-nicotinic acetylcholine receptor (alpha 7-nAChR). The electrostatic binding of cND-alpha-BTX was mediated by the negative charge of the cND and the positive charge of the alpha-BTX in physiological pH conditions. Sodium dodecyl sulfate-polyacrylamide gel analysis and matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI/TOF-MS) spectra displayed that alpha-BTX proteins were conjugated with cND particles via non-covalent bindings. The green fluorescence of the cND particles combining with the red fluorescence of tetramethylrhodamine-labeled a-BTX presented a yellow color at the same location, which indicated that alpha-BTX proteins were conjugated with cND particles. Xenopus laevis's oocytes expressed the human alpha 7-nAChR proteins by microinjection with alpha 7-nAChR mRNA. The cND-alpha-BTX complexes were bound to alpha 7-nAChR locating on the cell membrane of oocytes and human lung A549 cancer cells analyzed by laser scanning confocal microscopy. The choline-evoked alpha 7-nAChR-mediated inward currents of the oocytes were blocked by cND-alpha-BTX complexes in a concentration-dependent manner using two-electrode voltage-clamp recording. Furthermore, the fluorescence intensity of cND-alpha-BTX binding on A549 cells could be quantified by flow cytometry. These results indicate that cND-conjugated alpha-BTX still preserves its biological activity in blocking the function of alpha 7-nAChR, and provide a visual system showing the binding of alpha-BTX to alpha 7-nAChR.
Bio-imaging of alpha-bungarotoxin binding to target cell by carboxylated nanodiamond
(2008-04-01)
Chao, Jui-I
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Liu, Kuang-Kai
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Chen, Mei-Fang
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Chen, Po-Yi
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Lee, Tony J. F.
Biocompatible and detectable carboxylated nanodiamond on human cell
(2007-08-15)
Liu, Kuang-Kai
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Cheng, Chia-Liang
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Chang, Chia-Ching
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Chao, Jui-I
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生物科技學系
Surface-modified carboxylated nanometre-sized diamond (cND) has been applied for the conjugation of biological molecules such as DNA and protein. In this study, we evaluated the biocompatibility and detection of cNDs and carbon nanotubes on human lung A549 epithelial cells and HFL-1 normal fibroblasts. Treatment with 5 or 100 nm cND particles, 0.1 - 100 mu g ml(-1), did not reduce the cell viability and alter the protein expression profile in lung cells; however, carbon nanotubes induced cytotoxicity in these cells. The cNDs particles were accumulated in A549 cells, which were observed by atomic force microscopy and laser scanning confocal microscopy. Both 5 and 100 nm cNDs particles exhibited the green fluorescence and were ingested into cells. Moreover, the fluorescence intensities were increased in cells via a concentration-dependent manner after treatment with 5 and 100 nm cNDs, which can be detected by flow cytometer analysis. The fluorescence intensities of 5 nm cNDs were relative higher than 100 nm cNDs in cells at equal concentration treatment. The observation demonstrated that cND-interacting with cell is detectable by a confocal microscope, flow cytometer and atomic force microscope. These nanoparticles may be useful for further biomedical applications based on the properties of uptake ability, detectability and little cytotoxicity in human cells.
Biocompatible evaluation of fluorescent nanodiamonds in neurons in vitro
(2014-04-01)
Kao, Jun-Wei
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Huang, Yung-An
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Chiang, Ming-Han
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Huang, Hou-Syun
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Liu, Kuang-Kai
Cancer cell labeling and tracking using fluorescent and magnetic nanodiamond
(2012-09-01)
Lien, Zhi-Yi
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Hsu, Tzu-Chia
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Liu, Kuang-Kai
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Liao, Wei-Siang
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Hwang, Kuo-Chu
"Nanodiamond, a promising carbon nanomaterial, develops for biomedical applications such as cancer cell labeling and detection. Here, we establish the nanodiamond-bearing cancer cell lines using the fluorescent and magnetic nanodiamond (FMND). Treatment with FMND particles did not significantly induce cytotoxicity and growth inhibition in HFL-1 normal lung fibroblasts and A549 lung cancer cells. The fluorescence intensities and particle complexities were increased in a time- and concentration-dependent manner by treatment with FMND particles in lung cancer cells; however, the existence of FMND particles inside the cells did not alter cellular size distribution. The FMND-bearing lung cancer cells could be separated by the fluorescent and magnetic properties of FMNDs using the flow cytometer and magnetic device, respectively. The FMND-bearing cancer cells were identified by the existence of FMNDs using flow cytometer and confocal microscope analysis. More importantly, the cell morphology, viability, growth ability and total protein expression profiles in the FMND-bearing cells were similar to those of the parental cells. The separated FMND-bearing cells with various generations were cryopreservation for further applications. After re-thawing the FMND-bearing cancer cell lines, the cells still retained the cell survival and growth ability. Additionally, a variety of human cancer types including colon (RKO), breast (MCF-7), cervical (HeLa), and bladder (BFTC905) cancer cells could be used the same strategy to prepare the FMND-bearing cancer cells. These results show that the FMND-bearing cancer cell lines, which reserve the parental cell functions, can be applied for specific cancer cell labeling and tracking. (C) 2012 Elsevier Ltd. All rights reserved."
Chasing nanodiamond in cell cycle progression
(2009-04-01)
Chao, Jui-I
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Liu, Kuang-Kai
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生物科技學系
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Department of Biological Science and Technology
Co-delivery of paclitaxel and cetuximab by nanodiamond enhances mitotic catastrophe and tumor inhibition
(2017-08-29)
Lin, Yu-Wei
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Raj, Emmanuel Naveen
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Liao, Wei-Siang
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Lin, Johnson
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Liu, Kuang-Kai
The poor intracellular uptake and non-specific binding of anticancer drugs into cancer cells are the bottlenecks in cancer therapy. Nanocarrier platforms provide the opportunities to improve the drug efficacy. Here we show a carbon-based nanomaterial nanodiamond (ND) that carried paclitaxel (PTX), a microtubule inhibitor, and cetuximab (Cet), a specific monoclonal antibody against epidermal growth factor receptor (EGFR), inducing mitotic catastrophe and tumor inhibition in human colorectal cancer (CRC). ND-PTX blocked the mitotic progression, chromosomal separation, and induced apoptosis in the CRC cells; however, NDs did not induce these effects. Conjugation of ND-PTX with Cet (ND-PTX-Cet) was specifically binding to the EGFR-positive CRC cells and enhanced the mitotic catastrophe and apoptosis induction. Besides, ND-PTX-Cet markedly decreased tumor size in the xenograft EGFR-expressed human CRC tumors of nude mice. Moreover, ND-PTX-Cet induced the mitotic marker protein phospho-histone 3 (Ser10) and apoptotic protein active-caspase 3 for mitotic catastrophe and apoptosis. Taken together, this study demonstrated that the co-delivery of PTX and Cet by ND enhanced the effects of mitotic catastrophe and apoptosis in vitro and in vivo, which may be applied in the human CRC therapy.
Combination of Aurora A and Haspin inhibitors enhances p53 expression and tumor growth inhibition in human colorectal cancer
(2016-04-01)
Chang, Yun-Hsuan
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Lin, Chien-I
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Chen, Zan-Chu
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Liu, Kuang-Kai
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Chao, Jui-I
Combination of Aurora A and Haspin inhibitors enhances p53 expression and tumor growth inhibition in human colorectal cancer
(2016-04-01)
Chang, Yun-Hsuan
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Lin, Chien-I
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Chen, Zan-Chu
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Liu, Kuang-Kai
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Chao, Jui-I
Covalent linkage of nanodiamond-paclitaxel for drug delivery and cancer therapy
(2010-08-06)
Liu, Kuang-Kai
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Zheng, Wen-Wei
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Wang, Chi-Ching
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Chiu, Yu-Chung
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Cheng, Chia-Liang
A nanoparticle-conjugated cancer drug provides a novel strategy for cancer therapy. In this study, we manipulated nanodiamond (ND), a carbon nanomaterial, to covalently link paclitaxel for cancer drug delivery and therapy. Paclitaxel was bound to the surface of 3-5 nm sized ND through a succession of chemical modifications. The ND-paclitaxel conjugation was measured by atomic force microscope and nuclear magnetic resonance spectroscopy, and confirmed with infrared spectroscopy by the detection of deuterated paclitaxel. Treatment with 0.1-50 mu g ml(-1) ND-paclitaxel for 48 h significantly reduced the cell viability in the A549 human lung carcinoma cells. ND-paclitaxel induced both mitotic arrest and apoptosis in A549 cells. However, ND alone or denatured ND-paclitaxel (after treatment with strong alkaline solution, 1 M NaOH) did not induce the damage effects on A549 cells. ND-paclitaxel was taken into lung cancer cells in a concentration-dependent manner using flow cytometer analysis. The ND-paclitaxel particles were located in the microtubules and cytoplasm of A549 cells observed by confocal microscopy. Furthermore, ND-paclitaxel markedly blocked the tumor growth and formation of lung cancer cells in xenograft SCID mice. Together, we provide a functional covalent conjugation of ND-paclitaxel, which can be delivered into lung carcinoma cells and preserves the anticancer activities on the induction of mitotic blockage, apoptosis and anti-tumorigenesis.
Delivery of paclitaxel and cetuximab by nanodiamond enhances mitotic catastrophe and tumor inhibition in colorectal cancer
(2016-04-01)
Lin, Yu-Wei
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Liao, Wei-Siang
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Lin, Johnson
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Liu, Kuang-Kai
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Cheng, Hsiao-Chun
Delivery of paclitaxel and cetuximab by nanodiamond enhances mitotic catastrophe and tumor inhibition in colorectal cancer
(2016-04-01)
Lin, Yu-Wei
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Liao, Wei-Siang
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Lin, Johnson
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Liu, Kuang-Kai
;
Cheng, Hsiao-Chun
Endocytic carboxylated nanodiamond for the labeling and tracking of cell division and differentiation in cancer and stem cells
(2009-09-01)
Liu, Kuang-Kai
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Wang, Chi-Ching
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Cheng, Chia-Liang
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Chao, Jui-I
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生物科技學系
Nanodiamond (ND) is carbon nanomaterial developing for biological applications in recent years. In this study, we investigated the location and distribution of 100 nm carboxylated ND particles in cell division and differentiation. ND particles were taken into cells by macropinocytosis and clathrin-mediated endocytosis pathways. However, the cell growth ability was not altered by endocytic ND particles after long-term cell culture for 10 days in both A549 lung cancer cells and 3T3-L1 embryonic fibroblasts. ND particles were equal separating into two daughter cells of cell division approximately. Finally, the cell retained a single ND's cluster in cytoplasm after sub-cultured for several generations. Interestingly, ND's clusters were carried inside of cell but without inducing damages after long-term cell culture. Moreover, ND particles did not interfere with the gene or protein expressions on the regulation of cell cycle progression and adipogenic differentiation. Together, these findings provide that endocytic ND particles are non-cytotoxic in cell division and differentiation, which can be applied for the labeling and tracking of cancer and stem cells. (c) 2009 Elsevier Ltd. All rights reserved.
Endocytic nanodiamond for the labeling and tracking of cancer cells
(2010-04-01)
Chao, Jui-I
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Liu, Kuang-Kai
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生物科技學系
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Department of Biological Science and Technology
Labeling of neuronal differentiation and neuron cells with biocompatible fluorescent nanodiamonds
(2014-05-16)
Hsu, Tzu-Chia
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Liu, Kuang-Kai
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Chang, Huan-Cheng
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Hwang, Eric
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Chao, Jui-I
Nanodiamond is a promising carbon nanomaterial developed for biomedical applications. Here, we show fluorescent nanodiamond (FND) with the biocompatible properties that can be used for the labeling and tracking of neuronal differentiation and neuron cells derived from embryonal carcinoma stem (ECS) cells. The fluorescence intensities of FNDs were increased by treatment with FNDs in both the mouse P19 and human NT2/D1 ECS cells. FNDs were taken into ECS cells; however, FNDs did not alter the cellular morphology and growth ability. Moreover, FNDs did not change the protein expression of stem cell marker SSEA-1 of ECS cells. The neuronal differentiation of ECS cells could be induced by retinoic acid (RA). Interestingly, FNDs did not affect on the morphological alteration, cytotoxicity and apoptosis during the neuronal differentiation. Besides, FNDs did not alter the cell viability and the expression of neuron-specific marker beta-III-tubulin in these differentiated neuron cells. The existence of FNDs in the neuron cells can be identified by confocal microscopy and flow cytometry. Together, FND is a biocompatible and readily detectable nanomaterial for the labeling and tracking of neuronal differentiation process and neuron cells from stem cells.
Microwave-assisted efficient conjugation of nanodiamond and paclitaxel
(2015-05-15)
Hsieh, Yi-Han
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Liu, Kuang-Kai
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Sulake, Rohidas S.
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Chao, Jui-I.
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Chen, Chinpiao
Nanodiamond has recently received considerable attention due to the various possible applications in medical field such as drug delivery and bio-labeling. For this purpose suitable and effective surface functionalization of the diamond material are required. A versatile and reproducible surface modification method of nanoscale diamond is essential for functionalization. We introduce the input of microwave energy to assist the functionalization of nanodiamond surface. The feasibility of such a process is illustrated by comparing the biological assay of ND-paclitaxel synthesized by conventional and microwave irradiating. Using a microwave we manage to have approximately doubled grafted molecules per nanoparticle of nanodiamond. (C) 2015 Elsevier Ltd. All rights reserved.
Nanometer-sized diamond particle as a probe for biolabeling
(2007-09-01)
Chao, Jui-I.
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Perevedentseva, Elena
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Chung, Pei-Hua
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Liu, Kuang-Kai
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Cheng, Chih-Yuan
A novel method is proposed using nanometer- sized diamond particles as detection probes for biolabeling. The advantages of nanodiamond's unique properties were demonstrated in its biocompatibility, nontoxicity, easily detected Raman signal, and intrinsic fluorescence from its natural defects without complicated pretreatments. Carboxylated nanodiamond's ( cND's) penetration ability, noncytotoxicity, and visualization of cND- cell interactions are demonstrated on A549 human lung epithelial cells. Protein- targeted cell interaction visualization was demonstrated with cND- lysozyme complex interaction with bacteria Escherichia coli. It is shown that the developed biomolecule- cND complex preserves the original functions of the test protein. The easily detected natural fluorescent and Raman intrinsic signals, penetration ability, and low cytotoxicity of cNDs render them promising agents in multiple medical applications.
Synthesis and biological assay of erlotinib analogues and BSA-conjugated erlotinib analogue
(2017-04-15)
Boobalan, Ramalingam
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Liu, Kuang-Kai
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Chao, Jui-I.
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Chen, Chinpiao
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生物科技學系
A series of erlotinib analogues that have structural modification at 6,7-alkoxyl positions is efficiently synthesized. The in vitro anti-tumor activity of synthesized compounds is studied in two non-small cell lung cancer (NSCLC) cell lines (A549 and H1975). Among the synthesized compounds, the iodo compound 6 (ETN-6) exhibits higher anti-cancer activity compared to erlotinib. An efficient method is developed for the conjugation of erlotinib analogue-4, alcohol compound, with protein, bovine serum albumin (BSA), via succinic acid linker. The in vitro anti-tumor activity of the protein attached erlotinib analogue, 8 (ETN-4-Suc-BSA), showed stronger inhibitory activity in both A549 and H1975 NSCLC cell lines. (C) 2017 Elsevier Ltd. All rights reserved.