基因體學研究在藥學上之應用
Nucleic Acid Structure
DNA Structure
nitrogenous bases
– A, T, G, C
pentose sugar
– deoxyribose
usually a double helix, composed of two
complementary strands
– base pairing rules
A with T
G with C
nucleoside = nitrogenous base
+ pentose sugar
phosphodiester bond
two polynucleotide
chains are antiparallel
RNA Structure
nitrogenous bases
– A, G, C, U (instead of T)
pentose sugar
– ribose
usually consists of single strand
– can coil back on itself
forms hair-shaped structures with
complementary base pairing and helical
organization
base pairing rules
– A with U
– G with C
Types of RNA
three types
– ribosomal RNA (rRNA)
– transfer RNA (tRNA)
– messenger RNA (mRNA)
differ from each other in function, site of
synthesis in eucaryotic cells, and
structure
The Central Dogma
Pattern of DNA Synthesis
semiconservative
– each parental
strand is conserved
– two parental
strands separate
and serve as
templates for
synthesis of new
strands
transcription
recognition site for ribosome
synthesis continues
until terminator
reached
E. coli ribosome
site where amino
acid is attached
complementary to
codon in mRNA
Aminoacyl-tRNA
amino acid
 rest of tRNA
initiation
codon
transpeptidation reaction
Pharmacogenomics
• prolonged muscle relaxation after
Suxamethonium v.s. inherited deficiency
of plasma cholinesterase
Hemolysis after antimalarial therapy v.s.
inherited level of erythrocyte glucose 6phosphate dehydrogenase
• peripheral neutropathy of isoniazid
v.s. inherited differences in acetylation
of isoniazid
• Pharmacogenetics
define the more narrow spectrum of inherited
differences in the drug metabolism and
disposition
• Genes are considered to be polymorphic when
variant alleles exist stably in the population
- Gene Polymorphism
• Gene locus
A specific place on a chromosome where
a gene is located
• Allele
One of the different forms of a gene
that can exist at a single locus
• Allele frequency
The proportion of all alleles of that gene
in the population that are of this specific
type
P
AA
Gametes
aa
X
a
A
Zygote
Locus
Aa
½A
½a
½A
¼ AA
¼ Aa
½a
¼ Aa
¼ aa
• Multiple allelle
A, B, C, D…. Etc.
Genetic variation at the galactosemia locus
• gene encodes galactose-1-phosphate uridyltransferase (GALT)
• recessive mutation results in inability to metabolize galactose
• causes mental retardation and death
• some protection afforded by complete removal of milk from the diet
• variant alleles exist in addition to several galactosemia (g) alleles
• spectrum of enzymatic activities indicates that “normal” individuals
do not all have the same enzymatic activity levels
Genotype
Frequency
G/G
G/D
G/LA
G/g
D/D
D/LA
LA/LA
D/g
LA/g
g/g
87.4%
7.5
3.7
0.9
0.16
0.16
0.04
0.04
0.02
0.0025
Enzyme
Activity
100
75
120
50
50
95
140
25
70
<5
Phenotype
Normal
Normal
Normal
Normal
Normal
Normal
Normal
Borderline
Normal
Galactosemia
Relative number of individuals
Population distribution of “normal” GALT activities
G/G
G/g
G/LA
G/D
g/g
0
20
40
60
80
100
120
1 40
Relative GALT enzyme activity
160
CLASSIFICATION
Phase I (Functionalization): Phase II (Conjugation):
Oxidation
Cytochrome P450(CYP)
Alcohol Dehydrogenase(ADH)
Monoamine Oxidase
Reduction
Cytochrome P450
Hydrolysis
Esterases
Amidases
Glucuronosyltransferases(GT)
N-Acetyltransferases(NAT)
Sulfotransferases (ST)
Methyltransferases (MT)
Glutathione Transferases(GST)
Amino Acid Transferases
• Single nucleotide polymorphism
- Common variations among DNA of individuals
Nature Biotechnology May 2000
Volume 18 Number 5 pp 505 - 508
The use of single-nucleotide Polymorphism
maps in pharmacogenomics
Jeanette J. McCarthy1 & Rolf Hilfiker2
1. Millennium Predictive Medicine, Inc., Cambridge, MA 02139.
2. Solvias AG, Basel, Switzerland.
• Metabolic polymorphism
• Polymorphism of target proteins (e.g.
receptors)
• Transporter polymorphism
Genetic Polymorphisms in Drug Metabolism and Disposition
• Pharmacogenomics
Refer to the entire spectrum of genes that
determine the drug behavior and sensitivity
Proteomics
Evaluation of Protein-Level
Gene Expression
proteome
– entire collection of proteins that an organism
produces
proteomics
– study of the proteome
Two-dimensional gel
electrophoresis
when coupled with mass spectrometry,
used to:
– determine mass of each protein
– determine amino acid composition or
sequence of each protein
– identify protein
Pandey et.al., Nature 405 (2000)
Proteomic analysis of the effect of
Antrodia camphorata extract on
human lung cancer A549 cell
 樟芝
– 別名「紅樟」、「紅樟芝」
– 學名為（Antrodia camphorata）
 使用樟芝最早的是原住民
– 受驚、風寒、中暑、頭疼、跌打損傷、中毒
等內臟任何疾病都使用樟芝，為原住民的
「法寶」
 樟芝子實體具有強烈沖鼻的樟樹香氣，它的外型呈板
狀或鐘狀，面為橘紅(黃)色，整面全有菌孔
 樟芝最早的研究文獻報告在1990年發表，歸類為
靈芝屬。
 1995年張東柱等人依樟芝子實體型態及真菌的培
養基特性，將樟芝重新命名為Antrodia cinnamomea
 1997年吳聲華等人，依據前兩次文獻將樟芝命名
Antrodia camphorata 。
 樟芝的生理活性物質以三帖類、固醇類、多醣體
等化合物為主
抗菌研究
樟芝的甲醇萃取物可以有效抑制金黃葡萄球菌
及鬚滄小芽癬生長，且對腸道弛緩運動、血小
板凝集作用以及多種腸道菌有抑制效果
樟芝菌絲多醣體具有抑制B型肝炎抗毒以及抑
制B型肝炎病毒核心抗原(HBe)的活性
Lee et.al., FEMS Microbiology Letters 209
(2002)
保肝作用
Song等人 (2002) 利用四氯化碳誘導大鼠急性肝
損傷，來測試樟芝菌絲體深層培養之發酵濾液乾
燥物(DMF)
–降低肝臟血清之發炎指標-GOT及GPT 之生化值
–在肝臟組織病理切片中發現，能降低CCl4對大鼠引
發之肝細胞發炎(inflammation) 及壞死(necrosis)
等肝損傷現象
對腫瘤細胞反應能力
抑制癌細胞
The effect of AC on the survival of A549 cells
Control
AC
The calpain inhibitor CP1B  MMP-mRNA
expression in the leukemic THP-1 cells (Popp
et. al., Biol Chem 384)
AC(-)
MMP-2
Actin
AC(+)
Human Galectin-1
 A beta-galactoside binding protein.
 Cell adhesion, cell proliferation, and cell
death.
 Tumor-immune privilege .
(Rubinstein et. al., Cancer cell 5,2004)
Tumor Cell
Proliferation
(Activated)
AC(-)
Galectin-1
AC(+)
Eukaryotic translation initiation factor 5A (eIF5A)
Lysine residue
hypusine
 Proliferation and transformation of eukaryotic
cells
 Highly expressed in tumor cells.
(Prostate,ovarian, lung adenocarcinomas )
Chen et. al. Proteomics 2003

Eukaryotic translation initiation factor 5A (eIF-
5A)
 down-regulation of hypusine synthesis
Carraglia et. al., Adv Exp Med Biol. 472
Human calcium-dependent protease (Calpain)
 Cell progression to apotosis
 v-src transformation (Carraglia et. al., Mol.
Cell. Biol. 22)
 Degradation of p53 (Kubbatut et. al., Mol.
Cell. Biol. 17)
 Calpain inhibitor 1  p53-dependent apotosis
(Atencio et. al., Cell Growth Differ. 11)
Annexin V
 A calcium-binding protein annexin V
 As inhibitor of protein kinase C
 Annexin V proteins are highly expressed in tumor
cells, including breast tumor, bladder cancer)
(Celis et. al., Mol. Cell. Proteomics 1)
Rho-GDP dissociation inhibitor 
 Regulate the Rho family proteins
 Down-regulation of Rho-GDP dissociation
inhibitor   induce the apotosis of lung
cancer H157 cell (MacKeigan et. al., Cancer Res.
63 )