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Research Interest:
Oncology, Cell Biology Our
recent studies focus on the effects of HPV-E6 oncogene
and other environmental factors on P53 and its downstream
genes regulation. In general, the p53 gene expression is
as important response checkpoint in cells during stress
environment. However, these phenomena did not appear in
IR treated HPV-16 E6 transfected TK6 lymphoblastoid cells
in which IR did not induce p53 gene expression and its
downstream gene expression. TK6-E6 cells were more resistant
to ionizing radiation (IR) in comparison with their parental
TK6 cells. The high-risk HPVs E6 protein is thought to
bind P53, promoting it degradation through the ubiquitin
pathway and thus abrogates its function.
Recently, we have found arsenite treatment
could restore p53 expression and its downstream p21 and mdm2
gene expression in TK6-E6 cells. Using RT-PCR analysis, it showed
that E6-mRNA decreased in dose-response manner from TK6-E6 cells
after sodium arsenite treatment. It might explain why the restoration
of p53, its downstream p21 and mdm2 gene expression in these
originated P53-abrogated TK6-E6 cells. Since more activated caspase-3
and apoptosis appeared in TK6-E6 cells in comparison with parental
cells after treatment, Whether more p53 expression (due to E6
down regulation) would affect arsenite-induced apoptosis or not
in cells ? What are the mechanisms in this arsenite induced apoptosis
(p53-dependant or p53-independent) ? To continue these approaches,
we will apply p53-dominant negative vector into TK6-E6 cells
and examine the arsenite-induced apoptosis in these cells and
their parental TK6 cells.
Cancer Biology, Geno-toxicology According
to our previous study in TK6 and TK6-E6 cells, we believed that
arsenite treatment in SiHa (E6 containing) cells will reduce
E6 gene expression and increase p53 and its downstream p21 and
mdm2 gene expression, in addition, it will increase p53-dependant
apoptosis also. We will also apply ionizing radiation (in contrast
with arsenite treatment) in same cells to the differential response
between IR and arsenite-induced p53-dependant response in SiHa
cells. On the other hand, we will use HA1004, a protein kinase
A inhibitor, to examine arsenite-induced p53-independent apoptosis.
Through this study, we may provide another possible strategy
to promote arsenite as another potential candidate for the therapeutic
purpose in HPV-positive cancer cells.
Geno-toxicology and Biotech. The
complete genome sequence (1.6 Mb, open reading frames 1590) of Helicobacter
pylori has been determined by TGIR and several university laboratories
in 1997. Although it is relatively small and suitable for a structural
genomics and proteomics studies, the relative not much studies had
been reported.-- only 3-4 proteins in proteomics or structural biology
had been determined according to PDB record. Recent genomic DNA microarray
study from other lab. provided the evidence that many acid-inducible
proteins were found in bacteria in pH 5.5 (in comparison to pH 7.2).
Following these results, we will choose several target genes and
construct them into suitable expression vectors for further protein
expression in E. coli or in other system (cooperate with sub-project
5). Large preparation in pure proteins will be used for for NMR and
X-ray crystallography [cooperated with others in this structure biology
team for this project]. We predict to clone at least 15 to 20 target
genes in this individual laboratory within three years from the Helicobacter
pylori genome and expect to obtain 3-5 purified proteins for use
in NMR and X-ray crystallography per year.
Selected Publications:
Wang, H.J., Huang, H.M., Chuang, Y.C., and Huang, H.C. (2008). Paclitaxel induces up-regulation of tissue factor in human aortic endothelial cells. International Immunopharmacology (accepted).
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Wang, H.J., Huang, H.C., Chuang, Y.C., and Huang, H.M*. (2008). Thrombin Induces the Expression of Tissue Factor in Human Aortic Endothelial Cells. Acta Cardiologica Sinica24, 151-6.
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Chu, C.H., Lai, Y.J., Huang, H.M., and Sun, Y.J. (2008). Kinetic and structural properties of triosephosphate isomerase from Helicobacter pylori. Proteins: Structure, Function, and Bioinformatics 71, 396-406.
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Lee, M.J., Huang, H.M., Lin, W., Yang, R.R., Liu, C.L., and Huang, C.Y. (2007). Activation of Helicobacter pylori inorganic pyrophosphatase and the importance of Cys16 in thermostability, enzyme activation and quaternary structure. Archives of Microbiology 188, 473-482.
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Lu, P.K., Tsai, J.Y., Chien, H.Y., Huang, H.M., Chu, C.H., and Sun, Y.J. (2007). Crystal structure of Helicobacter pylori spermidine synthase: A Rossmann-like fold with a distinct active site. Proteins-Structure Function and Bioinformatics 67, 743-754.
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Chao, T.C., Huang,H.M., Tsai, J.Y., Huang, C.Y., and Sun, Y.J. (2006). Crystal Structure of Inorganic Pyrophophatase from the Pathogenic Bacterium Helicobacter pylori. PROTEINS: Structure, Function, and Bioinformatics. 65, 670–680.
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Lee, M.J., Liu, C.H., Wang, S.Y., Huang, C.T., and Huang, H.* (2006).Characterization of the Soj/Spo0J Chromosome Segregation Proteins and Identification of Putative parS Sequences in Helicobacter pylori. Biochemical and Biophysical Research Communications. 342, 744-750.
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Lee, M.J., Huang, C.Y., Sun, Y.J., and Huang, H*. (2005). Cloning and characterization of spermidine synthase and its implication in polyamine biosynthesis in Helicobacter pylori strain 26695. Protein Expression and Purification 43, 140-148.
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Ho, C.E., Tseng, F.G., Lin, S.C., Su, C.J., Liu, Z.Y., Yu, R.J., Chen, Y.F., Huang, H.M., and Chieng, C.C. (2005). Characterization of the surface tension and viscosity effects on the formation of nano-liter droplet arrays by an instant protein micro stamper. Journal of Micromechanics and Microengineering 15, 2317-2325.
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P.-K. Lyu, S.-Y. Chien, J.-Y. Tsai, C.-T. Fong,
M. J. Lee, H. M. Huang, Y.-J. Sun Crystallization and Preliminary
X-ray Diffraction Analysis of Spermidine Synthase from Helicobacter
Pylori, Acta Crystallography D 60, 2067-2069, 2004.
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Suh-Chin Wu., Haimei Huang, Chih-Chien Lin. Expression
and functional characterization of Helicobacter pyloricatalase
from baculovirus-infected insect cells. Enzyme and Microbial
Technology 35 (2004) 482–487
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S. C. Lin, F. G. Tseng, H. M. Huang, Yu-Feng Chen,
Y. C. Tsai, C. E. Ho, and C. C. Chieng , “Simultaneous Immobilization
of Protein Microarrays by a Micro Stamper with Back-Filling Reservoir'”, Sensors and Actuators, B: Chemical, 99(1): 174-185, 2004.
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Fangang Tseng, Shih-Chang Lin, Da-Jen Yao, Haimei
Huang, and Ching-Chang Chieng,“Technical Aspects of Protein Microarrays
and Nanoarrays” in “Protein Microarrays” ISBN: 0-7637-3127-7,
edited by Mark Schena, Jones and Bartlett Publishers (Sudbury,
MA), Chapter 16, pp.305-338, 2004
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H. M. Huang SY, Chen TT, Chen JC, Chiou CL,
Huang TM. Cisplatin restores p53 function and enhances the radiosensitivity
in HPV16 E6 containing cells Journal of Cellular Biochemistry 91,756-765 (2004)
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F. G. Tseng, S. C. Lin, H. M. Huang, C. Y. Huang,
and C. C. Chieng, Protein Micro Arrays Immobilized By m-Stamps
And Protein Wells On PhastgelR Pad Sensors and Actuators B 83,
22-29 (2002).
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Fan-Gang Tseng, Haimei Huang, Chang-Sheng Liu,
Chau-Yuh Huang, Ching-Chang Chieng, Size Effect on Surface Tension
and Contact Angle between Protein Solution and Silicon Compound,
PC and PMMA Substrates. Microscale Thermophysical Engineering 6, 31-53 (2002).
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Ruey-Hwang Chou and Haimei Huang. Activation of
p53 tumor suppressor pathway in cervical carcinoma cells by sodium
arsenite. Biochemical and Biophysical Research Communication 293, 298-306 (2002).
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Sun, N. K., Kamarajan, P., Huang, H.M., Chao,
C. C.-K. Chao, Restoration of UV sensitivity in UV-resistant
HeLa cells by antisense-mediated depletion of damaged DNA-binding
protein 2 (DDB2) FEBS Letters 25739, 1-5 (2002).
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Ruey-Hwang Chou and Haimei Huang*. Sodium arsenite
suppresses human papillomavirus 16 E6 gene and enhances apoptosis
in E6-transfected human lymphoblastoid cells. Journal of Cellular
Biochemistry 84, 615-624 (2002).
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Lin SC, Tseng FG, Huang HM, Huang CY, Chieng CC.
Microsized 2D protein arrays immobilized by micro-stamps and
micro-wells for disease diagnosis and drug screening. Fresenius
J Anal Chem. 371(2), 202-208 (2001).
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曾繁根, 黃海美, 林世章, 黃朝裕, 錢景常, 沾印式蛋白質微陣列晶片研製, 科儀新知 23(5),
14-23 (2000).
- Haimei Huang, Liu , Y. C., Shyu, Y. J. The Modulation
studies of Arsenite induced Genotoxicity. Chinese Public Health
J. 18(6), 174-185 (1999).
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