1
Supplementary Material 2
In the primary screen of the anti-cancer traditional Chinese medicinal herbs and derived
monomers using vasculogenic tumor cells as a model, we found that the extract of Coptis
Chinensis and derived JH and other 4 monomers showed inhibitory effects on the
proliferation of human melanoma cells as follows:
1. The extraction of Coptis chinensis: The active components of Coptis chinensis were
prepared as follows: For the ethanol extract, the whole plant of Coptis chinensis (100 g)
was chopped and extracted three times with 80% ethanol (reflux, 2 h each time), and then
the ethanol extracts were combined and evaporated in a vacuum to yield an ethanol
extract 9.68 g (yield 9.7%). For the water extract, the whole plant of Coptis chinensis
(100 g) was chopped and extracted for three times with 100 ℃ water (reflux, 2 h each
time), and then the water extracts were combined and evaporated in a vacuum to yield an
water extract 17.08 g (yield 17.1%).
2. Next, we examined the effect of extracts from Coptis chinensis on the proliferation of
human melanoma C8161 and A375 cells by Alamarblue. The result showed that both
ethanol and water extracts of Coptis chinensis had anti-melanoma cell proliferative effect,
and C8161 cells were more sensitive to the extracts as compared with A375 cells
(Supplementary Table 2).
Supplementary Table 2. Effect of ethanol and water extracts of Coptis chinensis on
human melanoma cell proliferation
extract (μg/ml)
ethanol extract
C8161
A375
water extract
C8161
A375
2
0.625
2.6%
1.5%
0.7%
0.1%
1.25
8.8%
4.9%
7.0%
4.7%
2.5
14.5%
8.0%
7.3%
7.0%
5
18.5%
16.6%
16.0%
11.9%
10
42.2%
35.2%
34.7%
27.1%
20
78.5%
53.4%
54.8%
32.4%
40
84.3%
81.7%
78.6%
62.9%
80
92.3%
96.8%
86.2%
93.2%
3. Next, we tested the effect of 5 monomers purified from Coptis chinensis on human
melanoma cell C8161 proliferation by Alamarblue, including obakulactone mognoflorine,
obakunone, ferulic acid, and Jatrorrhizine hydrochloride (JH). As shown in
Supplementary Tables 3 and 4, JH exerted anti-melanoma proliferative effect, but others
not obviously. Therefore, we focused on JH.
Supplementary Table 3. Effect of several monomers from Coptis chinensis on human
melanoma cell C8161 proliferation by Alamarblue
Drug (μM)
obakulactone
mognoflorine
obakunone
ferulic acid
JH
1.25
-2.2%
4.4%
-1.6%
-6.8%
-0.9%
2.5
-0.9%
-0.4%
-1.4%
0.5%
2.2%
3
5
0.5%
2.0%
1.1%
2.2%
3.3%
10
0.9%
-1.0%
2.6%
3.6%
9.7%
20
9.0%
7.6%
2.7%
4.3%
21.8%
40
-1.4%
0.8%
1.8%
4.4%
47.4%
80
-4.0%
3.3%
0.6%
2.6%
71.2%
160
3.6%
8.9%
10.3%
12.0%
87.9%
Supplementary Table 4. Effect of several monomers from Coptis chinensis on human
melanoma cell A375 proliferation by Alamarblue
Drug (μM)
obakulactone
mognoflorine
obakunone
ferulic acid
JH
1.25
1.2%
3.2%
-1.3%
0.3%
-5.9%
2.5
-1.4%
2.5%
-2.4%
0.4%
-0.1%
5
0.0%
1.5%
-2.5%
1.5%
-1.0%
4
10
0.9%
1.2%
-6.1%
-1.1%
3.0%
20
0.7%
1.4%
1.1%
-4.9%
3.9%
40
0.2%
6.2%
5.5%
1.8%
9.0%
80
-1.1%
3.1%
-4.8%
-0.1%
27.0%
160
-2.9%
7.9%
8.6%
11.4%
55.8%
Supplementary Table 5. Effect of JH on human solid cancer cells proliferation by
Alamarblue
Number
1
2
3
4
5
6
Cell line
Eahy926
C8161
HepG2
HL-60
H9
HEL
Cancer cells
human umbilical vein endothelial cells
human metastatic melanoma cells
human hepatic carcinoma cells
human original myelogenous leukemia cells
human T lymphoma cells
human erythroleukemia cell line
IC50 (μM)
45.6
47.4
47.4
50.7
51.1
51.1
5
7
8
9
10
12
13
CCRF
Jurkat
NB-4
Daudi
MDA-MB231
LM3
SGC-7901
14
CaSki
15
16
17
18
19
20
21
22
23
24
25
26
27
28
29
30
31
32
33
34
35
36
37
38
39
40
41
Dami
U-2 OS
U251
ECV-304
HMY2-CIR
PaTu8988
KP-N-NS
RD
NAMALWA
Caki-1
My-5
Saos-2
K562
SW1990
AGS
SW480
MG-63
UC-CUC
U-87MG
HO-8910PM
HCCC-9810
U937
A172
N87
Bxpa-3
MCF-7
HT-29
11
human acute leukemia T lymphoma cells
human T lymphoma cell leukemia cells
human leukemia cell line
human megakaryocytic leukemia cells
51.7
54.4
56.3
57.9
human breast cancer cells
57.9
human hepatic carcinoma cells
human gastric cancer cells
human cervical cancer with intestinal metastasis
cells
human megakaryocytic leukemia cells
human osteosarcoma cells
human glioma cells
human umbilical vein endothelial cells
human B lymphoblastoid cells
human pancreatic cancer cells
human adrenal neuroblastoma cells
human embryonal rhabdomyosarcoma cells
human Burkitt's lymphoma cells
human renal adenocarcinoma cell line
human myeloma cell line
human osteosarcoma cells
human chronic myeloid leukemic cell line
human pancreatic cancer cells
human gastric cancer cells
human colon cancer cells
human osteosarcoma cells
human monocytic leukemia cells
human glioma cells
human high metastatic ovarian cancer cells
human bile duct cell type liver cells
human tissue cell lymphoma cells
human glioblastoma cells
human gastric cancer cells
human pancreatic cancer cells
human breast adenocarcinoma cell line
human colon carcinoma cell line
61.1
61.9
62.5
63.1
63.8
66.2
67.4
69.5
75.2
75.6
77.5
78.7
79.5
81.4
81.7
82.2
83.2
83.2
83.6
84.2
85.7
86.0
89.0
94.7
94.9
98.2
99.1
99.3
101.0
114.1
6
Supplementary Figure 1. Effect of JH on the body weight of nude mice with Matrigel
plugs