肿瘤干细胞 肿瘤干细胞 Cancer Stem Cells
Cancer Stem Cells
AntiAnti--Cancer Stem Cell ( C S C )Cancer Stem Cell ( C S C )
Hoechst 33342 & Hoechst 33342 & Side Population (SP) CellsSide Population (SP) Cells
Hoechst 33342: dsDNA-binding dye
UV Excitation Emission350 nm 460 nm
FilterBlue Red
450/20 nm 674 nm(Goodell MA et al_1996)
SPSP
Do selected TCM herbs as components of the clinically used anti-cancer formula, with reported anti-tumour bioactivity, specifically target cancer stem cells(CSC)?
1. Examination of the crude extracts of selected anti-cancer herbal medicines
2. Subsequent fractionation to obtain pure compounds that specifically target CSC
CSC in vitro system => Changes in the
proportion of the Side Population as surrogate CSC
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
61.5
Solvent control (80% EtOH) for HM1 to HM5. Dissolved in DMSO.
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
44.3
3.78
10.9
3.32
10.9%
Standard 3: Negative controls: solvent control
10-1 100 101 102 103
FL3 LOG: PI
10-1
100
101
102
103
F
L
1
L
O
G
:
F
I
T
C
0.47 2.37
73.124
a b
c
a: DMSO 0.1% control
b: Chemotherpeutics Taxol 0.5 ug/ml
C: Thalidomide 20 ug/ml
16% 73%
35%
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
14.6
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
55.6
PI0 1000 2000 3000 4000
UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
14.6
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
55.6
PI0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
55.6
PI
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
20.2
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
70.1
PI0 1000 2000 3000 4000
UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
20.2
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
70.1
PI0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
70.1
PI
a. 0.2% DMSO,
b. 0.0002% DMSO
14.6%
20.2%
Positive vsnegative controls
Chemo Taxol
0.0001 ug/ml
Chemo Taxol
0.001 ug/ml
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
19.8
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000F
S
C
-
A
62.2
PI0 1000 2000 3000 4000
UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
19.8
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000F
S
C
-
A
62.2
PI0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000F
S
C
-
A
62.2
PI
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
20
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
59.7
PI0 1000 2000 3000 4000
UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
20
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
59.7
PI0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
59.7
PI
19.8%
20.0%
Neg control
Chemo Taxol
0.01 ug/ml
Chemo Taxol
0.1 ug/ml
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
18.2
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000F
S
C
-
A
54.2
PI0 1000 2000 3000 4000
UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
18.2
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000F
S
C
-
A
54.2
PI0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000F
S
C
-
A
54.2
PI
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
21.1
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
8.57
PI0 1000 2000 3000 4000
UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
21.1
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
8.57
PI0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
8.57
PI
18.2%
21.1%
Neg control
Propolis from different geographic areas have different chemical profiles
1620
2428
32
900720
540360
180
Mass (a.m.u.)
0.0
3.0
1.5
Intensity (log)
A
b
s
o
r
b
a
n
c
e
(
%
)
0
50
100
Retention Time (mi
n)
P1 P2 P5 P4 STD P3
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
6.36
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
43.8
1.82
16.5
1.82
HM2 and HM4
at 100 ug/mL,
had high killing effect on the cancer cells,
but did not preferentially killed the SP.
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
1.95
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
43.1
3.08
18.3
1.91
HM2
HM crude extracts that killed the cancer cells with no killing preference for the SP
HM4
16.5%
18.3%
Herbal Medicine (HM) crude extract
that selectively targets and kills SP
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
42.5
HM 1 at 100 ug/mL, selectively reduced SP%
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
21.9
8.96
0.83
0.11
0.83%
POSITIVE control
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
2.74
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
33.2
3.17
0.49
1.38
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
25.1
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
14.5
10.5
0.43
0.062
b.) Fractions from HM1 that selectively killed SP
Fraction 6 (F6) and fraction 7(F7) at 100 ug/mL, selectively killed the SP
F6
F7
0.49%
0.43%
POSITIVE control
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
59.4
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
39.1
4.02
12.3
2.07
C2: Solvent control for fraction 1
0 5000 104 50000105
B575 D-A
0
1000
2000
3000
4000
F
S
C
-
A
53.6
0 1000 2000 3000 4000UV670 A-A
0
1000
2000
3000
4000
U
V
4
4
0
_
B
-
A
34.5
3.74
15.3
2.32
C3: Solvent controls for fractions 4, 6 & 7
C2
C3
Solvent controls for fractions obtained from HM. Dissolved in DMSO.
12.3%
15.3%
SOLVENTcontrols
Peer-reviewed RESEARCH FUNDINGS
Anthony Rothe Memorial Trust (Australia)
University of Sydney Cancer Research Fund
International Myeloma Foundation (USA)
Multiple Myeloma Research Fund (USA)
Cancer Institute NSW Career Development Fund (Australia)
NSW Cancer Council Project Fund (Australia)
Australia-China Special Fund
Australian National Institute of Complementary Medicine
George Li, Kei Fan, Iyad Ahmed, Nantiga Virgona, Bhagwant Sekhon, Tetsuo Yamagishi, Rebecca Roubin
Gouyi Mo, Suilin Mo, Daniel Sze, Shihong Yang, Celine Loh, Kristin Muller
Cancer Immunology Group,
Faculty of Pharmacy,
University of Sydney