Kuete et al. BMC Complementary and Alternative Medicine 2014, 14:340
http://www.biomedcentral.com/1472-6882/14/340
RESEARCH ARTICLE
Open Access
Cytotoxicity of four Aframomum species
(A. arundinaceum, A. alboviolaceum, A. kayserianum
and A. polyanthum) towards multi-factorial drug
resistant cancer cell lines
Victor Kuete1,3*, Patrick Y Ango2, Samuel O Yeboah4, Armelle T Mbaveng3, Renameditswe Mapitse4, Gilbert DWF Kapche5,
Bonaventure T Ngadjui4 and Thomas Efferth1*
Abstract
Background: The search for natural products as potential cytotoxic agents has yielded promising candidates.
However multidrug resistance (MDR) is still a major hurdle for patients receiving chemotherapy. In the present
study, we evaluated the cytotoxicity of the methanol extracts of four dietary Aframomum plant species (A.
arundinaceum, A. alboviolaceum, A. kayserianum and A. polyanthum) against nine sensitive and MDR cancer cell
lines. We have also identified the bioactive constituents of A. arundinaceum.
Methods: The cytotoxicity of the methanol extracts of the above plants was determined using a resazurin
reduction assay. Chromatographic techniques were used to isolate the constituents of A. arundinaceum.
Results: A preliminary experiment on leukemia CCRF-CEM cells at 40 μg/mL showed that the extracts from A.
kayserianum and A. alboviolaceum as well as the isolated compounds namely compounds aframodial (1), 8
(17),12-labdadien-15,16-dial (2), galanolactone (3), 1-p-menthene-3,6-diol (6) and 1,4-dimethoxybenzene (7) were
less active, inducing more than 50% growth of this cell line contrary to A. polyanthum and A. arundinaceum extracts,
galanals A (4) and B (5), naringenin (8) and kaempferol-3,7,4’-trimethylether (9). The IC50 values below or around 30 μg/
mL were recorded with A. arundinaceum extract against eight of the nine tested cancer cell lines. This extract as well as
compound 8 displayed IC50 values below 40 μg/mL towards the nine tested cancer cell lines whilst A. polyanthum
extract, compounds 4, 5 and 9 showed selective activities. Collateral sensitivity (hypersensitivity) was observed
with A. arundinaceum extract towards leukemia CEM/ADR5000 cells and glioblastoma U87MG.ΔEGFR compared to
their respective sensitive counterparts CEM/CEM and U87MG.
Conclusion: The results of this study provide evidence of the cytotoxicity selected Aframomum species as well as
a baseline information for the potential use of Aframomum arundinaceum in the fight against drug sensitive and
otherwise drug-resistant cancers.
Keywords: Aframomum, Cameroon, Cancer, Cytotoxicity, Multidrug resistant, Zingiberaceae
* Correspondence: kuetevictor@yahoo.fr; efferth@uni-mainz.de
1
Department of Pharmaceutical Biology, Institute of Pharmacy and
Biochemistry, University of Mainz, 55128 Mainz, Germany
Full list of author information is available at the end of the article
© 2014 Kuete et al.; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative
Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and
reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain
Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article,
unless otherwise stated.
Kuete et al. BMC Complementary and Alternative Medicine 2014, 14:340
http://www.biomedcentral.com/1472-6882/14/340
Background
Chemotherapy remains the major treatment of cancers
but often fails due to cells multidrug resistance (MDR)
[1,2]. MDR is displayed by many cancer cells to withstand
increasingly higher doses of antineoplastic compounds [3].
Investigation for naturally occurring molecules as potential cytotoxic drugs has yielded promising candidates
[3-7]. However, MDR is still considered a major hurdle for
patients receiving chemotherapy [8,9]. Various Cameroonian dietary plants including those from the family
Zinziberaceae are used in traditional medicine to manage various ailments [5,10-13]. The genus Aframomum,
belonging to the Zingiberaceae family have 40 species
and is most common in tropical and subtropical regions [14]. Twenty species are found in Cameroon,
where they are widely used as spices and in traditional
medicine [14]. The Seeds of Aframomum arundinaceum
K. Schum are used as laxative and as anti-helmintic. The
fresh juice of the rhizomes is used against body odor. The
rhizomes are used against toothache and the crushed
seeds against fungal infections [10]. The decoction of the
leaves Aframomum melegueta K. Schum together with the
leaves of Momordica charantia and Sorghum arundinaceum cereal in local dry gin (alcohol) is recommended to
be taken one dose daily against cholera [15]. Several
Aframomum species such as Aframomum angustifolium,
A. danielli, A. sanguineum, and A. sulcatum are also traditionally used to treat fevers in Africa [16], and recently,
the antiplasmodial activity of some labdanes from A. sceptrum and A. latifolium was demonstrated [17]. The antibacterial activities of Aframomum kayserianum [12] and
Aframomum polyanthum [13] were also reported on
Gram-negative multidrug-resistant phenotypes. The cytotoxicity of other Afromomum species such as A. citratum
and A. melegueta towards leukemia CCRF-CEM and
ADR5000 cell lines was also reported [5]. The present
study was designed to investigate the cytotoxicity of
four dietary Aframomum species commonly used as
spices in Cameroon, Aframomum alboviolaceum (Ridl.)
K. Schum, A. arundinaceum (Oliver & Hanbury) K. Schum,
Aframomum kayserianum K. Schum and Aframomum
polyanthum K. Schum towards sensitive and multi-factorial
drug resistant cancer cell lines. The study was extended
to the identification of the bioactive constituents of
A. arundinaceum.
Page 2 of 7
(Yaoundé, Cameroon) where voucher specimens were
deposited under the reference numbers 11704/SFR/
CAM (A. arundinaceum), 34888/HNC (A. alboviolaceum), 18884/SRFC (A. kayserianum) and 3981/SRFK
(A. polyanthum). The air dried fruits of A. kayserianum,
A. polyanthum (100 g) and A. arundinaceum (3000 g)
as well as the roots of A. alboviolaceum (100 g) were
powdered and macerated with methanol for 48 h at
room temperature. The methanol extract was concentrated in vacuo to give 18.7 g, 21.2 g, 25.3 and 100 g of
the crude extracts of A. kayserianum, A. polyanthum,
A. alboviolaceum and A. arundinaceum respectively.
The extracts were then conserved at 4°C until further
use.
Isolation of compounds from Aframomum arundinaceum
Crude extract of A. arundinaceum (100 g) was successively extracted with petroleum ether, chloroform and
methanol at room temperature. The petroleum ether
fraction (25 g) was column chromatographed on 100 g
of silica gel (Merck, 0.040-0.063 mm) using hexane and
hexane-choloroform mixture with increasing polarity.
Fractions of 300 mL were collected, concentrated, and
pooled on the basis of their thin layer chromatography
(TLC) profile. The obtained fractions (frs) directly
afforded a yellow oil (1; frs 4 to 9; 30 mg) and amorphous powders 2 ( frs 13 to 16; 25 mg), 3 (frs 20 to 27;
40 mg), 4 (frs 30 to 33) and 5 (frs 36 to 38; 10 mg). The
chloroform extract (20 g) was also column chromatographed on 250 g of silica gel (Merck, 0.040-0.063 mm)
using hexane (Hex) and mixture of hexane-chloroform
(Hex-CHCl3). Fractions of 400 mL were collected, concentrated and pooled after TLC analysis to give five subfractions (sub-frs A-E).
Sub-fraction B (Hex-CHCl3; 10 to 25; 6 g) was subjected to column chromatography (CC) to afford a white
crystal (6; 20 mg). Sub-fraction C (8.0 g) obtained with
Hexane-CHCl3 4:6 was subjected to CC (silica gel 60,
50 g) and eluted with Hex-CHCl3 mixtures of increasing
polarity to give 6 new sub-fractions (C1-C6). Subfraction C4 obtained with Hex-CHCl3 6:4. afforded a yellow oil (7; 10 mg). Sub-fraction C5 (Hex-CHCl3 4:6) and
C6 (Hex-CHCl3 8:2) were repeatedly filtered through
Sephadex LH-20 (CHCl3-MeOH 7:3) to give yellow
powders, 8 (sub-frs 3 to 6; 10.0 mg) and 9 (sub-frs 15 to
19; 15 mg).
Methods
Plant material and extraction
The tested Aframomum species, A. alboviolaceum, A.
kayserianum and A. polyanthum were purchased from
Bafoussam local market (West region of Cameroon) in
January 2012. Aframomum arundinaceum was collected
in Yaoundé (Centre region) in March 2012. The plants
were further identified at the National Herbarium
General procedure
Aluminum sheet pre-coated with silica gel 60 F254 nm
(Merck) was used for thin layer chromatography; the
spots were visualized using both ultraviolet light (254
and 366 nm) and 50% H2SO4 spray reagent. NMR spectra were recorded on a Bruker Avance 300 (Billerica,
Kuete et al. BMC Complementary and Alternative Medicine 2014, 14:340
http://www.biomedcentral.com/1472-6882/14/340
MA, USA) at 300 MHz (1H) and 75 MHz (13C), with the
residual solvent peaks as internal references. Mass spectra
were recorded with API QSTAR pulsar mass (Milford,
MA, USA). Melting points (m.p) were recorded using a
Stuart Scientific (Redhill, Surrey, UK) melting point apparatus (SMP1) and are uncorrected. The chemical
structures of the compounds were confirmed by comparing with reference data from available literature
(Figure 1).
Chemicals
Doxorubicin 98.0% were provided by the University
Pharmacy of the Johannes Gutenberg University (Mainz,
Germany) and dissolved in PBS (Invitrogen, Eggenstein,
Germany) at a concentration of 10 mM. Geneticin >98%
(72.18 mM; Sigma-Aldrich, Munich, Germany).
Page 3 of 7
Cell cultures
The cell lines used the present work, their origins and
their treatments were previously reported [18,19]. They include drug-sensitive CCRF-CEM and multidrug-resistant
P-glycoprotein over-expressing CEM/ADR5000 leukemia
cells [20-22], the MDA-MB-231-pcDNA3 breast cancer
cells and its resistant subline MDA-MB-231-BCRP clone
23) [23], the HCT116 (p53+/+) colon cancer cells and its
knockout clones HCT116 (p53-/-), the U87MG glioblastoma cells and its resistant subline U87MG.ΔEGFR,
HepG2 hepatocarcinoma cells and AML12 normal hepatocytes [6,19,24]. The CCRF-CEM and CEM/ADR5000
leukemia cells were maintained in RPMI 1640 medium
(Invitrogen) supplemented with 10% fetal calf serum in
a humidified 5% CO2 atmosphere at 37°C. Sensitive and
resistant cells were kindly provided by Dr. Axel Sauerbrey
(Department of Pediatrics, University of Jena, Jena,
Figure 1 Chemical structures of compounds isolated from the fruit of Aframomum arundinanceum K Schum. 1: aframodial; 2: 8(17),
12-labdadien-15,16-dial; 3: galanolactone; 4: galanal A; 5: galanal B; 6: 1-p-menthene-3,6-diol; 7: 1,4-dihydroxybenzene; 8: naringenin;
9: kaempferol-3,7,4’-trimethylether.
Kuete et al. BMC Complementary and Alternative Medicine 2014, 14:340
http://www.biomedcentral.com/1472-6882/14/340
Page 4 of 7
Germany). The generation of the resistant subline was
previously described [6,19,24]. Breast cancer cells, transduced with control vector (MDA-MB-231-pcDNA3) or
with cDNA for the breast cancer resistance protein BCRP
(MDA-MB-231-BCRP clone 23), were maintained under
standard conditions as described above for CCRF-CEM
cells. Human wild-type HCT116 (p53+/+) colon cancer
cells as well as knockout clones HCT116 (p53-/-) derived by homologous recombination were a generous
gift from Dr. B. Vogelstein and H. Hermeking (Howard
Hughes Medical Institute, Baltimore, MD). Human glioblastoma multiforme U87MG cells (non-transduced) and
U87MG cell line transduced with an expression vector
harboring an epidermal growth factor receptor (EGFR)
gene with a genomic deletion of exons 2 through 7
(U87MG.ΔEGFR) were kindly provided by Dr. W. K.
Cavenee (Ludwig Institute for Cancer Research, San
Diego, CA). MDA-MB-231-BCRP, U87MG.ΔEGFR and
HCT116 (p53-/-) were maintained in DMEM medium
containing 10% FBS (Invitrogen) and 1% penicillin (100
U/mL)-streptomycin (100 μg/mL) (Invitrogen) and were
continuously treated with 800 ng/mL and 400 μg/mL
geneticin, respectively. Human HepG2 hepatocellular
carcinoma cells and normal AML12 heptocytes were
obtained from the American Type Culture Collection
(ATCC, USA). The above medium without geneticin
was used to maintain MDA-MB-231, U87MG, HCT116
(p53+/+), HepG2 and AML12 cell lines. The cells were
passaged twice weekly. All experiments were performed
with cells in the logarithmic growth phase.
wavelength of 544 nm and an emission wavelength of
590 nm. Each assay was done at least two times, with six
replicates each. IC50 values represent the sample’s concentrations required to inhibit 50% of cell proliferation
and were calculated from a calibration curve by linear
regression using Microsoft Excel [5,6].
Resazurin reduction assay
The cytotoxicity of the studied samples was performed
by resazurin reduction assay as we previously described
[6,18,19,24-26]. Briefly, adherent cells at 1x104 cells were
allowed to attach overnight and then treated with different studied samples. Samples were preliminary tested at
40 μg/mL (extract and isolated compounds) and doxorubicin (20 μg/mL) against the sensitive leukemia CCRFCEM cell line and those inducing less than 50% growth
proliferation were further tested for IC50 determinations
towards all the studied cell lines. For suspension cells,
aliquots of 2 × 104 cells per well were seeded in 96-wellplates in a final volume of 200 μL. Extracts and compounds were prior diluted in DMSO and tested in a final
concentration below 0.1% (A final concentration of 0.1%
DMSO was used as negative control and did not show
any effect on cell growth). The tested concentrations
ranges were 0.16 μg/mL to 40 μg/mL for crude extracts
and isolated compounds and 0.08 μg/mL to 20 μg/mL
for doxorubicin. After 72 h incubation and a resazurin
(Sigma-Aldrich, Schnelldorf, Germany) staining, fluorescence was measured on an Infinite M2000 Pro™ plate
reader (Tecan, Crailsheim, Germany) using an excitation
Results and discussion
The structures of the compounds isolated from Aframomum arundinaceum were established using spectroscopic analysis, especially, NMR spectra in conjunction
with 2D experiments, COSY, HMQC, HMBC, and direct comparison with published information and with
authentic specimens obtained in our research group for
some cases. The compounds isolated from the fruits of
A. arundinaceum (Figure 1) were identified as Aframodial C20H30O3 (1; m/z 318.2) [27], 8(17),12-labdadien15,16-dial C20H30O2 (2; m/z 302.2) [17], galanolactone
C20H30O3 (3; m/z 318.2) [27], galanal A C20H30O3 (4;
15 mg, m/z 318.2) [28], and galanal B C20H30O3 (5; m/z
318.2) [29], 1-p-menthene-3,6-diol C10H18O2 (6; m/z 170.1;
m.p:165-167°C) [30], 1,4-dihydroxybenzene C6H6O2 (7;
m/z 110.0) [31], naringenin C15H12O5 (8; m/z 272.0;
245-248°C) [32] and kaempferol-3,7,4’-trimethylether
C18H16O6 (9; m/z 328.0; 157-158°C) [33]. The cytotoxicity of compounds 1-9 as well as the crude extracts was
determined towards drug sensitive and resistant cancer
cell lines.
In a preliminary investigation of the four studied Aframomum species and compounds isolated from A. arundinaceum, we tested a single concentration of 40 μg/mL
for each sample and 20 μg/mL for doxorubicin against
the sensitive CCRF-CEM leukemia cell line (Figure 2).
The extracts from A. kayserianum and A. alboviolaceum
were less active and induced respectively 50.33% and
54.36% growth proliferation of CCRF-CEM cells. Compounds 1, 2, 3, 6 and 7 also induced more than 50%
growth of this cell line. The extracts from A. polyanthum (36.28%) and A. arundinaceum (24.68%) as well
as compounds 4 (47.78%), 5 (49.81%), 8 (38.49%) and 9
(39.58%) displayed less than 50% growth proliferation of
CCRF-CEM cells. The IC50 values of the above samples
were further determined on nine cancer cell lines, including both sensitive and MDR phenotypes (Table 1).
Aframomum. arundinaceum extract as well as compound 8 and doxorubucin induced less than 50% proliferation of all tested cancer cell lines, with IC50 values
below 40 μg/mL. A. polyanthum extract, compounds 9,
4 and 5 showed selective activities, the IC50 values
<40 μg/mL being obtained on 5/9, 4/9, 2/9 and 1/9
tested cell lines respectively (Table 1). According to the
National Cancer Institute (USA), 30 μg/mL is the upper
IC50 limit considered promising for purification of a
crude extract [34]. We therefore, tested a slightly higher
Kuete et al. BMC Complementary and Alternative Medicine 2014, 14:340
http://www.biomedcentral.com/1472-6882/14/340
Page 5 of 7
Figure 2 Growth percentage (%) of leukemia CCRF-CEM cancer cell line treated with plant extracts and isolated compounds at 40 μg/
mL and doxorubicin (20 μg/mL). 1: aframodial; 2: 8(17),12-labdadien-15,16-dial; 3: galanolactone; 4: galanal A; 5: galanal B; 6: 1-p-menthene-3,
6-diol; 7: 1,4-dihydroxybenzene; 8: naringenin; 9: kaempferol-3,7,4’-trimethylether.
concentration of 40 μg/mL in our preliminary assay. Also,
the IC50 threshold value of 4 μg/ml or 10 μM [35,36] after
48 and 72 h incubations has been set to identify good
cytotoxic compounds. Considering these thresholds, the
IC50 values below or around 30 μg/mL were recorded with
A. arundinaceum extract against eight of the nine tested
cancer cell lines (Table 1) explaining why it was considered further for purification. Nonetheless, the extract from
A. polyanthum also showed activities with IC50 values
<30 μg/mL on four of the nine tested cancer cell lines.
Though Compound 8 was active on all the tested cancer
cell lines, no IC50 below 4 μg/ml was recorded, the lowest
values being 7.86 μg/mL against CEM/ADR5000 cells.
Interestingly, none of the selected extracts and compounds was more toxic towards AML12 normal hepatocytes (IC50 > 40 Mg/mL) than cancer cell lines,
suggesting their good selectivity. Importantly, collateral
sensitivity (hypersensitivity) was also observed with A.
arundinaceum extract towards CEM/ADR5000 cells
(degree of resistance of 0.76) and U87MG.ΔEGFR (degree of resistance of 0.95) compared to their respective
sensitive counterparts CEM/CEM and U87MG. This
extract was also more active against hepatocarcinoma
HepG2 as compared to AML12 normal hepatocytes, confirming its selectivity to cancer cells (Table 1). Despite the
fact that compound 8 showed moderate activities, it also
Table 1 Cytotoxicity of the studied Aframomum extracts, compounds and doxorubicin towards sensitive and
drug-resistant cancer cell lines and normal cells as determined by the resazurin assay
Studied samples, IC50 values (μg/mL)a and degree of resistance (in braket)
Cell lines
Aframomum species
Compounds from A. arundinaceum
Doxorubucin
A. polyanthum
A. arundinaceum 4
5
8
9
CCRF-CEM
20.37 ± 3.10
18.08 ± 0.98
19.81 ±
2.01
12.20 ± 1.87
18.38 ± 2.04
CEM/ADR5000
28.16 ± 1.24 (1.38) 13.73 ± 1.02 (0.76) - (>2.31)
- (>2.02)
7.86 ± 0.74 (0.64)
18.22 ± 1.18 (0.99) 195.12 ± 14.30
(1772)
-
33.79 ± 2.38
-
9.51 ± 1.03
MDA-MB-231BCRP
30.24 ± 2.18 (0.89) 30.66 ± 3.17 (1.02) 27.99 ± 2.39
(<0.70)
-
18.12 ± 2.01 (1.91) 33.14 ± 2.64
(<0.83)
7.83 ± 0.01 (7.11)
HCT116 p53+/+
-
23.06 ± 2.21
-
-
13.65 ± 1.11
1.43 ± 0.02
HCT116 p53
-
27.38 ± 1.92 (1.19) -
-
13.86 ± 0.94 (1.02) 36.74 ± 2.31
(<0.82)
4.06 ± 0.04 (2.84)
U87MG
-
36.70 ± 2.12
-
-
29.81 ± 1.88
1.06 ± 0.03
U87MG.ΔEGFR
20.59 ± 1.87
(<0.51)
24.42 ± 1.95 (0.67) -
-
18.02 ± 1.34 (0.60) -
6.11 ± 0.04 (5.76)
HepG2
-
23.15 ± 1.97
(<0.58)
-
-
23.46 ± 1.95
(<0.59)
-
1.41 ± 0.12 (<0.04)
AML12
-
-
-
-
-
-
-
a
-
0.11 ± 0.01
MDA-MB-231
-/-
29.98 ± 1.86
17.32 ± 1.96
-
-
1.10 ± 0.01
The degree of resistance was determined as the ratio of IC50 value in the resistant divided by the IC50 in the sensitive cell line; AML12 was used as the
corresponding resistant counterpart for HepG2. 4: galanal A; 5: galanal B; 8: naringenin; 9: kaempferol-3,7,4’-trimethylether; (-): >40 μg/mL.
Kuete et al. BMC Complementary and Alternative Medicine 2014, 14:340
http://www.biomedcentral.com/1472-6882/14/340
displayed better collateral sensitivity of MDR cell lines
compared to doxorubicin. The use of natural products to
fight multidrug resistance is an attractive strategy in
chemotherapy [37-39]. P-gp-expressing CEM/ADR5000
as well as p53 knock out HCT116 (p53-/-) and BCRPexpressing U87MG.ΔEGFR cells were less cross-resistant
towards the best samples namely A. arundinaceum and
compound 8 than towards the positive drug, doxorubicin,
highlighting their possible therapeutic potential in the
fight against multidrug resistance. This report also highlights the importance of the plants of the genus Aframomum as potential source of cytotoxic compounds. The
results obtained collaborate with previous investigations.
In effect, Aframomum melegueta previously inhibited the
proliferation of the leukemia ADR5000 cell lines with a reported IC50 value of 7.80 μg/mL [5]. Also, naringenin (8)
has shown cytotoxicity in various human cancer cell lines
and induced apoptosis via a transient induction of
caspase-3/CPP32 activity, in the human promyeloleukemia cell line HL-60 [40-42]. The moderate cytotoxicity of
galanals A (4; IC50 of 18 μM or 5.62 μg/mL) and B (5;
IC50 of 32 μM or 12.21 μg/mL) towards human T lymphoma Jurkat cells was also reported [29].
Conclusions
Finally, this work provides further evidence of the cytotoxic potential of Aframomum species and highlights the
good activity of Aframomum arundinaceum on sensitive
and drug-resistant cancer cell lines. Bioactive constituents of this plant include galanals A and B, naringenin
and kaempferol-3,7,4’-trimethylether. Aframomum arundinaceum could be explored in more detail in the future
to develop novel anticancer drugs against sensitive and
resistant phenotypes.
Competing interests
The authors declare that they have no competing interests.
Authors’ contributions
VK, PYA and ATM carried out the study; PYA, ATM, SOY, RM, GDWF and BTN
contributed to plant’s collection, compound’s isolation and/or identification.
VK and TE designed the experiments. VK wrote the manuscript; TE
supervised the work, provided the facilities for the study. All authors read
and approved the final manuscript.
Acknowledgements
VK is very grateful to the Alexander von Humboldt foundation for an
18 months’ fellowship in Germany through the “Georg Foster Research
Fellowship for Experienced Researcher” program; PYA is grateful to the
Network of Analytical and Bioassay Services in Africa (NABSA) for a 2-months
maintenance grant to the University of Botswana.
Author details
1
Department of Pharmaceutical Biology, Institute of Pharmacy and
Biochemistry, University of Mainz, 55128 Mainz, Germany. 2Department of
Chemistry, Faculty of Science, University of Botswana, Francistown, Botswana.
3
Departments of Biochemistry, Faculty of Science, University of Dschang,
Dschang, Cameroon. 4Departments of Organic Chemistry, Faculty of Science,
University of Yaoundé I, Yaoundé, Cameroon. 5Department of Chemistry,
Page 6 of 7
Higher Teachers’ Training College, University of Yaoundé I, Yaoundé,
Cameroon.
Received: 4 July 2014 Accepted: 16 September 2014
Published: 19 September 2014
References
1. Jemal A, Bray F, Center MM, Ferlay J, Ward E, Forman D: Global cancer
statistics. CA-Cancer J Clin 2011, 61(2):69–90.
2. Chen Z, Zhang L, Xia L, Jin Y, Wu Q, Guo H, Shang X, Dou J, Wu K, Nie Y,
Fan D: Genomic analysis of drug resistant gastric cancer cell lines by
combining mRNA and microRNA expression profiling. Cancer Lett 2014,
350(1–2):43–51.
3. Marchini S, Marrazzo E, Bonomi R, Chiorino G, Zaffaroni M, Weissbach L,
Hornicek FJ, Broggini M, Faircloth GT, D’Incalci M: Molecular
characterisation of two human cancer cell lines selected in vitro for their
chemotherapeutic drug resistance to ET-743. Eur J Cancer 2005,
41(2):323–333.
4. Kuete V, Mbaveng AT, Tsaffack M, Beng VP, Etoa FX, Nkengfack AE, Meyer JJ,
Lall N: Antitumor, antioxidant and antimicrobial activities of Bersama
engleriana (Melianthaceae). J Ethnopharmacol 2008, 115(3):494–501.
5. Kuete V, Krusche B, Youns M, Voukeng I, Fankam AG, Tankeo S, Lacmata S,
Efferth T: Cytotoxicity of some Cameroonian spices and selected
medicinal plant extracts. J Ethnopharmacol 2011, 134(3):803–812.
6. Kuete V, Sandjo L, Nantchouang Ouete J, Fouotsa H, Wiench B, Efferth T:
Cytotoxicity and modes of action of three naturally occuring xanthones
(8-hydroxycudraxanthone G, morusignin I and cudraxanthone I) against
sensitive and multidrug-resistant cancer cell lines. Phytomedicine 2013,
21(3):315–322.
7. Kuete V, Sandjo LP, Kwamou GM, Wiench B, Nkengfack AE, Efferth T:
Activity of three cytotoxic isoflavonoids from Erythrina excelsa and
Erythrina senegalensis (neobavaisoflavone, sigmoidin H and
isoneorautenol) toward multi-factorial drug resistant cancer cells.
Phytomedicine 2014, 21(5):682–688.
8. Goldstein LJ, Galski H, Fojo A, Willingham M, Lai S-L, Gazdar A, Pirker R,
Green A, Crist W, Brodeur GM, Lieber M, Cossman J, Gottesman MM, Pastan
I: Expression of multidrug resistance gene in human cancers. J Natl
Cancer Inst 1989, 81(2):116–124.
9. Kuwazuru Y, Yoshimura A, Hanada S, Ichikawa M, Saito T, Uozumi K,
Utsunomiya A, Arima T, Akiyama S-I: Expression of the multidrug transporter, P-glycoprotein, in chronic myelogenous leukaemia cells in blast
crisis. Brit J Haematol 1990, 74(1):24–29.
10. Tane P, Tatsimo S, Ayimele G, Connolly J: Bioactive metabolites from
Aframomum species. In 11th NAPRECA Symposium Book of Proceedings.
Antananarivo, Madagascar: NAPRECA; 2005:214–223.
11. Dzoyem JP, Guru SK, Pieme CA, Kuete V, Sharma A, Khan IA, Saxena AK,
Vishwakarma RA: Cytotoxic and antimicrobial activity of selected
Cameroonian edible plants. BMC Complement Altern Med 2013, 13:78.
12. Djeussi DE, Noumedem JA, Seukep JA, Fankam AG, Voukeng IK, Tankeo SB,
Nkuete AH, Kuete V: Antibacterial activities of selected edible plants
extracts against multidrug-resistant Gram-negative bacteria. BMC
Complement Altern Med 2013, 13(1):164.
13. Seukep JA, Fankam AG, Djeussi DE, Voukeng IK, Tankeo SB, Noumdem JA,
Kuete AH, Kuete V: Antibacterial activities of the methanol extracts of
seven Cameroonian dietary plants against bacteria expressing MDR
phenotypes. Springerplus 2013, 2:363.
14. Thomas D, Thomas J, Bromley W, Mbenkum F: Korup ethnobotany survey,
final report to: The World Wide Fund for Nature. In Weyside Park,
Godalming, Surrey, UK: Penda House; 1989.
15. Ndukwu B, Ben-Nwadibia N: Ethnomedicinal aspects of plants used as spices
and condiments in the Niger delta area of Nigeria. Port Harcourt, Nigeria:
University of Port Harcourt PMB; 2010.
16. Iwu M: Handbook of African Medicinal Plants. Boca Raton, FL: CRC Press;
1993.
17. Duker-Eshun G, Jaroszewski JW, Asomaning WA, Oppong-Boachie F, Olsen
CE, Christensen SB: Antiplasmodial activity of labdanes from Aframomum
latifolium and Aframomum sceptrum. Planta Med 2002, 68(7):642–644.
18. O’Brien J, Wilson I, Orton T, Pognan F: Investigation of the Alamar Blue
(resazurin) fluorescent dye for the assessment of mammalian cell
cytotoxicity. Eur J Biochem 2000, 267(17):5421–5426.
Kuete et al. BMC Complementary and Alternative Medicine 2014, 14:340
http://www.biomedcentral.com/1472-6882/14/340
19. Kuete V, Tchakam PD, Wiench B, Ngameni B, Wabo HK, Tala MF, Moungang ML,
Ngadjui BT, Murayama T, Efferth T: Cytotoxicity and modes of action of four
naturally occuring benzophenones: 2,2′,5,6′-tetrahydroxybenzophenone,
guttiferone E, isogarcinol and isoxanthochymol. Phytomedicine 2013,
20(6):528–536.
20. Kimmig A, Gekeler V, Neumann M, Frese G, Handgretinger R, Kardos G,
Diddens H, Niethammer D: Susceptibility of multidrug-resistant human
leukemia cell lines to human interleukin 2-activated killer cells. Cancer
Res 1990, 50(21):6793–6799.
21. Efferth T, Sauerbrey A, Olbrich A, Gebhart E, Rauch P, Weber HO, Hengstler
JG, Halatsch ME, Volm M, Tew KD, Ross DD, Funk JO: Molecular modes of
action of artesunate in tumor cell lines. Mol Pharmacol 2003,
64(2):382–394.
22. Gillet J, Efferth T, Steinbach D, Hamels J, de Longueville F, Bertholet V,
Remacle J: Microarray-based detection of multidrug resistance in human
tumor cells by expression profiling of ATP-binding cassette transporter
genes. Cancer Res 2004, 64(24):8987–8993.
23. Doyle LA, Yang W, Abruzzo LV, Krogmann T, Gao Y, Rishi AK, Ross DD: A
multidrug resistance transporter from human MCF-7 breast cancer cells.
Proc Natl Acad Sci U S A 1998, 95(26):15665–15670.
24. Kuete V, Sandjo L, Wiench B, Efferth T: Cytotoxicity and modes of action of
four Cameroonian dietary spices ethno-medically used to treat Cancers:
Echinops giganteus, Xylopia aethiopica, Imperata cylindrica and Piper
capense. J Ethnopharmacol 2013, 149(1):245–253.
25. Kuete V, Fankam AG, Wiench B, Efferth T: Cytotoxicity and modes of action
of the methanol extracts of six Cameroonian medicinal plants against
multidrug-resistant tumor cells. Evid Based Complement Alternat Med 2013,
2013:285903.
26. Kuete V, Tankeo SB, Saeed ME, Wiench B, Tane P, Efferth T: Cytotoxicity and
modes of action of five Cameroonian medicinal plants against
multi-factorial drug resistance of tumor cells. J Ethnopharmacol 2014,
153(1):207–219.
27. Kamdem Wabo H, Tane P, Connolly J: Diterpenoids and sesquiterpenoids
from Aframomum arundinaceum. Biochem Syst Ecol 2006, 34:603–605.
28. Morita H, Itokawa H: Cytotoxic and antifungal diterpenes from the seeds
of Alpinia galanga. Planta Med 1988, 54(2):117–120.
29. Miyoshi N, Nakamura Y, Ueda Y, Abe M, Ozawa Y, Uchida K, Osawa T:
Dietary ginger constituents, galanals A and B, are potent apoptosis
inducers in Human T lymphoma Jurkat cells. Cancer Lett 2003,
199(2):113–119.
30. Bousetla A, Konuklugil B, Bouacida S, Zellagui A, Rhouati S, Akkal S:
Phytochemical study of Algerian Foeniculum vulgare Mill (Apiaceae).
Der Pharmacia Lettre 2013, 5(6):9–11.
31. Rogerson FSS, Azevedo Z, Fortunato N, de Freitas VAP: 1,3Dimethoxybenzene, a newly identified component of port wine.
J Sci Food Agric 2002, 82(11):1287–1292.
32. Nilupa R, Lalith J, Noriyuki H, Yoshinori F: Chemical constituents of the
fruits of Artocarpus altilis. Biochem Syst Ecol 2007, 36:323–325.
33. Pizzolatti M, Verdi L, Brighente I, Neiva T, Schripsema J, Braz Filho R:
Anticoagulant effect and constituents of Baccharis illinita. Nat Prod
Commun 2006, 1(1):37–42.
34. Suffness M, Pezzuto J: Assays related to cancer drug discovery. London:
Academic Press; 1990.
35. Boik J: Natural compounds in cancer therapy. Minnesota USA: Oregon
Medical Press; 2001.
36. Brahemi G, Kona FR, Fiasella A, Buac D, Soukupova J, Brancale A, Burger AM,
Westwell AD: Exploring the structural requirements for inhibition of the
ubiquitin E3 ligase breast cancer associated protein 2 (BCA2) as a
treatment for breast cancer. J Med Chem 2010,
53(7):2757–2765.
37. Efferth T: The human ATP-binding cassette transporter genes: from the
bench to the bedside. Curr Mol Med 2001, 1(1):45–65.
38. Gottesman MM, Ling V: The molecular basis of multidrug resistance in
cancer: the early years of P-glycoprotein research. FEBS Lett 2006,
580(4):998–1009.
39. Gillet JP, Efferth T, Remacle J: Chemotherapy-induced resistance by
ATP-binding cassette transporter genes. Biochim Biophys Acta 2007,
1775(2):237–262.
40. Chen YC, Shen SC, Lin HY: Rutinoside at C7 attenuates the apoptosisinducing activity of flavonoids. Biochem Pharmacol 2003,
66(7):1139–1150.
Page 7 of 7
41. Kanno S, Shouji A, Asou K, Ishikawa M: Effects of naringin on hydrogen
peroxide-induced cytotoxicity and apoptosis in P388 cells. J Pharmacol
Sci 2003, 92(2):166–170.
42. Wang BD, Yang ZY, Wang Q, Cai TK, Crewdson P: Synthesis,
characterization, cytotoxic activities, and DNA-binding properties of the
La(III) complex with Naringenin Schiff-base. Bioorg Med Chem 2006,
14(6):1880–1888.
doi:10.1186/1472-6882-14-340
Cite this article as: Kuete et al.: Cytotoxicity of four Aframomum species
(A. arundinaceum, A. alboviolaceum, A. kayserianum and A. polyanthum)
towards multi-factorial drug resistant cancer cell lines. BMC Complementary
and Alternative Medicine 2014 14:340.
Submit your next manuscript to BioMed Central
and take full advantage of:
• Convenient online submission
• Thorough peer review
• No space constraints or color figure charges
• Immediate publication on acceptance
• Inclusion in PubMed, CAS, Scopus and Google Scholar
• Research which is freely available for redistribution
Submit your manuscript at
www.biomedcentral.com/submit