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Inorganic Chemistry Communications 73 (2016) 138141

Contents lists available at ScienceDirect

Inorganic Chemistry Communications

journal homepage: www.elsevier.com/locate/inoche

Short communication

Potential anti-cancer activity of a novel Bi(III) containing


thiosemicarbazone derivative
Ruizhuo Ouyang a,,1, Yang Yang a,1, Xiao Tong b, Yaoqin Yang b, Huihong Tao b, Tianyu Zong a, Kai Feng a,
Pengpeng Jia a, Penghui Cao a, Ning Guo a, Haizhou Chang a, Shuang Zhou b,, Yuqing Miao a,
a
University of Shanghai for Science and Technology, Shanghai 200093,China
b
Cancer Institute, Tongji University School of Medicine, Shanghai 200092, China

a r t i c l e i n f o a b s t r a c t

Article history: By mixing 5-bromo-2-furaldehyde, phenylthiosemicarbazide and Bi(CH3COO)3, a new amorphous bismuth-con-
Received 20 July 2016 taining complex, BiL2Cl3, was synthesized under aqueous conditions via a one-pot method and characterized
Received in revised form 3 October 2016 with some analytical techniques, where L was produced by dehydration between 5-bromo-2-furaldehyde,
Accepted 7 October 2016
phenylthiosemicarbazide. The obtained complex effectively inhibited the proliferation and migration of A549
Available online 11 October 2016
and H460 lung cancer cells, induced cell apoptosis and displayed fairly low cytotoxicity to normal human lung
Keywords:
broblast (HLF). Compared with cisplatin, BiL2Cl3 displayed better anti-cancer activity against tumor cells and
Bi(III) complex lower toxicity to normal cell. Therefore, BiL2Cl3 with potent anti-cancer activity may be a viable drug candidate
Thiosemicarbazone derivative in anti-cancer therapies.
Anti-cancer ability 2016 Elsevier B.V. All rights reserved.
Lung cancer
A549 and H460

1. Introduction signicant cytotoxic activity against various cancer cell lines [18]. As
their biological activities mostly depend on the difdent groups at N-4 po-
Due to the unusually low toxicity [1], bismuth compounds are pref- sition, more and more substituted thiosemicarbazones have been devel-
erably selected to replace the traditional triple therapy with bismuth- oped with improved biological activities [19]. Since nitrogen containing
containing quadruple rescue therapy for the treatment of Helicobacter heterocyclic molecules constitute the largest portion of chemical entities,
pylori-infected gastropathy [25]. Their insolubility in neutral aqueous the vital part of many natural biologically active pharmaceuticals to
solutions like biological uids results in the remarkably low toxicity. enhance the survivability [20], a large number of metal-tridentate
However, low pH environment increases the solubility of bismuth com- thiosemicarbazones complexes derived from -N-heterocyclic
pounds. As we know, pH value of tumor site is relatively lower than that carboxaldehydes were reported to exhibit signicant biological activities
of normal tissues. Accordingly, bismuth (both Bi(III) and Bi(V)) com- [7,21,22]. The coordination of organic compounds with metal ions can
plexes as cancer drugs have been developed and used for tumor treat- cause drastic change in the biological properties of the ligand and the
ment [611], which likely accumulate in tumor site, but show low metal ion moieties [23]. A new Bi(III) complex was accordingly synthe-
content in blood [12,13]. Their bioactivities in the treatment of various sized using 5-bromo-2-furaldehyde and phenylthiosemicarbazide as li-
gastrointestinal disorders, antitumor, antimicrobial, and antibacterial gands via a one-pot method. The aim of this work was to evaluate the
activities have been reported [14,15]. anti-cancer effect of this novel Bi(III) containing thiosemicarbazone deriv-
Thiosemicarbazones show good versatility as ligands because of the p- ative through a series of in vitro bioassays.
delocalisation of charge and the congurationally exibility of their mo- By mixing Bi(CH3COO)3, 5-bromo-2-furaldehyde and 4-
lecular chain, demonstrating high antitumor potential [16,17]. 3- phenylthiosemicarbazide, Bi(III) coordinated with 5-bromo-2-
Aminopyridine-2-carboxaldehyde-thiosemicarbazone is a well-known furaldehyde followed by dehydration between NH2 of 4-
representative compound as a promising anti-cancer agent due to its phenylthiosemicarbazide and CHO of 5-bromo-2-furaldehyde,
forming thiosemicarbazone (Fig. S1). And the subsequent coordination
of Bi(III) with thiosemicarbazone further accelerated the dehydration
Corresponding authors. between 4-phenylthiosemicarbazide and 5-bromo-2-furaldehyde, ulti-
E-mail addresses: ouyangrz@usst.edu.cn (R. Ouyang), shuangzhou@tongji.edu.cn
(S. Zhou), yqmiao@usst.edu.cn (Y. Miao).
mately producing as many complex as possible. The obtained Bi com-
1
R. Ouyang and Y. Yang contributed equally to this work and should be considered as plex was characterized by energy dispersive spectroscopy (EDS), 1H
co-rst authors. NMR and X-ray diffraction (XRD) (Figs. S2S4). A549 and H460, highly

http://dx.doi.org/10.1016/j.inoche.2016.10.020
1387-7003/ 2016 Elsevier B.V. All rights reserved.
R. Ouyang et al. / Inorganic Chemistry Communications 73 (2016) 138141 139

Table 1
Half maximum inhibitory concentration (IC50) of A549, H460 and HLF with BiL2Cl3 obtain-
ed via the MTT assay.
a
IC50 (M)

Cancer cell lines A549 H460 HLF


Bi 114.62 4.33 133.87 5.30 143.75 6.72
BiL2Cl3 16.41 0.93 20.04 1.28 117.16 5.96
Cisplatin 39.78 2.23 46.21 2.72 97.16 4.31
a
Data were shown as the mean SD (standard deviation) of ve independent
experiments.

invasive lung cancer cell lines, and normal human lung broblast (HLF)
were chosen as the model cell lines. The antiproliferative effects of
BiL2Cl3 on these cells were evaluated using the MTT assay (Table 1).
The results showed that BiL2Cl3 exhibited potent anti-cancer activity
against cancer cell lines and fairly low toxicity to normal cell, while
Bi(CH3COO)3 alone was inactive, which indicated that the coordination
of Bi(III) with ligands 4-phenylthiosemicarbazide and 5-bromo-2-
furaldehyde exerted a pronounced effect to the anti-cancer activity of
BiL2Cl3 against cells A549 and H460, and would likely present a good
therapeutic prole. Compared with cisplatin, BiL2Cl3 displayed better
anti-cancer activity against tumor cell lines and lower toxicity to normal
cell. In addition, both dose and time dependences of BiL2Cl3 were ob-
served for the inhibition of A549 and H460 cell growth (Fig. 1). With
more BiL2Cl3 used, obvious inhibition was found at rst beginning of
cancer cells treatment. However, the cancer cells A549 and H460 appar-
ently kept growing after being treated over 20 h when less than 20 M
BiL2Cl3 was used. The stable and excellent inhibition rate of BiL2Cl3
against two cancer cells was obtained at the concentration higher than
20 M, but bio-toxicity would inevitably occur when the concentration
is too high. Accordingly, 20 M of BiL2Cl3 was used in vitro as the opti-
mum dosage for the lung cancer cells treatment.
Colony formation assay can measure the cell proliferation capacity in
the living environment. The smaller the colony formation rate is, the
stronger the drugs prevent the cell proliferation. According to the in
Fig. 2. The effects of BiL2Cl3 on A549 and H460 single cell proliferation shown by colony
vitro cytotoxicity assay, 20 M BiL2Cl3 was used through the whole col- formation assay. Data were shown as the mean SD of ve independent experiments.
ony formation assay with Bi(CH3COO)3 at the same concentration for
comparison (Fig. 2A and B). Compared with control, colony formation
rate was found to be higher than 50% for both A549 and H460 after group. On the contrary, after being treated with BiL2Cl3, the wound
being treated with Bi(CH3COO)3, while no obvious colony formation size did not obviously change smaller for both cells at any treatment pe-
(less than 10%) occurred for both cancer cells after BiL2Cl3 treatment. riod (Fig. 3C and D), demonstrating that BiL2Cl3 possessed excellent
The results exhibit that the prepared BiL2Cl3 displayed excellent anti- anti-cell migration activity. Therefore, the prepared BiL2Cl3 signicantly
cell-proliferation activity in lung cancer cells. blocked the migration of A549 and H460 cells.
Cell migration plays an important role in tumor metastasis [24], Abrogation of apoptotic pathways is frequently found in malignant
wound-healing assays were thus performed to determine the effects cells, while most of the readily available cytotoxic chemical-based
of BiL2Cl3 on migration of A549 and H460 cells. Fig. 3A and B showed drugs mediate their effects by inducing apoptosis in cancer cells. Here,
that both cells clearly migrated to the wound area in both control BiL2Cl3 was found to be able to kill A549 and H460 cells predominantly
(DMSO) and Bi(CH3COO)3-treated experiment groups with treatment through the induction of apoptosis. To further analyze the mechanism
time, and the wound size greatly decreased after 24 h, showing a cell of BiL2Cl3-induced cell death, the involvement of apoptosis in cell
migration rate of higher than 70% for H549 and 90% for H460 in each death was investigated using Annexin-VFITC/PI double staining assays.

Fig. 1. Dose and time dependent drug efcacy studies of BiL2Cl3 on lung cancer cell lines A) A549 and B) H460. Data were shown as the mean SD of ve independent experiments.
140 R. Ouyang et al. / Inorganic Chemistry Communications 73 (2016) 138141

Fig. 3. The effects of BiL2Cl3 on A) A549 and B) H460 cell motility conducted with wound healing assay. The change in migration rate of BiL2Cl3 with time for C) A549 and D) H460 cell lines,
using contorl and Bi(CH3COO)3 as comparision. Data were shown as the mean SD of ve independent experiments.

Compared with control and Bi(CH3COO)3 treatment, exposure of either Hoechst staining (Fig. S5), the majority of the cells displayed condensed
A549 or H460 to BiL2Cl3 dramatically triggered apoptotic cell death and highly uorescent nuclei, a characteristic morphology of cells un-
(Fig. 4A and B). The population of necrotic cells (Annexin-VFITC-nega- dergoing apoptosis. Therefore, the cytotoxic effect of BiL2Cl3 on both
tive and PI-positive cells) was always as low as that of the control. As A549 and H460 cancer cells were attributed to the apoptosis. The results
seen in Fig. 4C and D, BiL2Cl3 induced 17.60 3.64% (A549) and of the ow cytometric analysis were consistent with those of the MTT
23.78 4.34% (H460) cell apoptosis, respectively, much higher than tests.
0.62 0.14% (A549) and 1.78 0.14% (H460) in the control group. Summary, a new amorphism thiosemicarbazone derivative, BiL2Cl3,
When A549 and H460 cells were incubated with BiL2Cl3 followed by was synthesized through a one-pot combinatorial metal three-

Fig. 4. Flow cytometry dot plots of A) A549 and B) H460: untreated cells (control), cells treated with Bi(CH3COO)3 and BiL2Cl3; the populations of C) A549 and D) H460 exposed to different
complexes with the same concentration. In all cases, the cells were exposed to the drug for 24 h at 37 C. FL1: the green channel of Annexin-VFITC; FL2: the red channel of PI. (For
interpretation of the references to color in this gure legend, the reader is referred to the web version of this article.)
R. Ouyang et al. / Inorganic Chemistry Communications 73 (2016) 138141 141

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