DOI: 10.1039/C5CC02149A
Page 3 of 4
Journal Name
ChemComm
COMMUNICATION
the same treatment (Fig. S20, ESI†). To further verify the function of anticancer drug CPT, and restore the fluorescence of CPT as well. In
DT-diaphorase on prodrug, A549 cells were pretreated with a DT- this way, the prodrug can fluorescently detect DT-diaphorase and
diaphorase inhibitor dicoumarol (20μM) for 30 min, then incubated realize real-time monitoring of drug release. The prodrug shows
with the prodrug (10 μM) for 2 h. From Fig. 2, we can see the cells much higher cytotoxicity towards DT-diaphorase overexpressed
exhibited negligible fluorescent emission upon pretreatment by cancer cells. This strategy may offer a approach for the development
dicoumarol. The results indicate that the fluorescent CPT can be of enzyme-catalyzed theranostic anticancer therapeutics.
released from the prodrug in DT-diaphorase overexpressed cancer
cells, which was further confirmed by flow cytometry analysis (Fig.
S21, ESI†). Moreover, we also investigated the fluorescent response
of the control drug to the intracellular DT-diaphorase in A549 cells
in the presence or absence of dicoumarol (Fig. S22, ESI†). The
control drug shows constant fluorescence in the cells, regardless of
the presence or absence of the DT-diaphorase inhibitor.
We acknowledge the financial support by NSFC (21474031,
21174040 and 21025415), and the National Key Basic
Research Program of China (Project No. 2013CB834702).
Notes and references
College of Materials Science & Engineering, State Key Lab of Luminescent
Materials & Devices,South China University of Technology, Guangzhou
510640, P. R. China. Fax: +86 20 22236363; Tel: +86 20 22236262; E-mail:
Additionally, cell viability experiment was carried out to
investigate the prodrug’s cytotoxicity towards two cell lines using
MTT assays (Fig. 3I). The prodrug shows strong cytotoxicity
towards DT-diaphorase overexpressed A549 cells, and the IC50
(concentration inhibiting cell growth to 50% of control) value is
1.179 μM. However, the cytotoxicity is much lower towards the
non-DT-diaphorase overexpressed cell L929 (IC50> 80 μM, ~90%
cell viability at 1.179 μM). Furthermore, the Annexin V-
FITC/propidium iodide (PI) method was performed to investigate
the apoptosis for two cell lines treated with the prodrug or CPT (Fig.
3II). The cells were first incubated with the prodrug or free CPT and
then stained with Annexin V-FITC/PI. The subsequent flow
cytometry analysis shows that the apoptotic percentage (including
the early and late apoptosis, Q2 and Q4 in Fig. 3 II) for A549 cells is
51.31% and 58.70% upon exposure to prodrug and CPT,
respectively. However, for the L929 cells, at the drug concentration
of as high as 20 μM, the percentage for apoptotic cells is only 7.58%
upon prodrug treatment and 37.32% upon CPT treatment. The
results also demonstrate the prodrug’s high selectivity to damage the
DT-diaphorase overexpressed cancer cells. Also, MTT assays for the
† Electronic Supplementary Information (ESI) available: [1H NMR
spectra, mass spectra, absorption spectra]. See DOI: 10.1039/b000000x/
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Fig. 3 (I) Cell viability profiles for A549 and L929 cell lines treated with the
prodrug of varied concentrations. (II) Representative percent distribution of
A549 and L929 cells analyzed by flow cytometry using annexin V-FITC/PI staining.
A549 cells were treated by the prodrug (5 μM) (A) and CPT (5 μM) (B) for 24 h,
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In conclusion, a DT-diaphorase-activatable theranostic prodrug
has been developed. In the presence of DT-diaphorase, the prodrug
undergoes cyclization and 1,6-elimination to release active
9
O. Redy and D. Shabat, J. Control. Release, 2012, 164, 276.
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