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ChemComm
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that the probe and BRACO19 compete the same binding target.
Moreover, the in vitro competition titrations demonstrated that
the probe bound to pu27 G4-DNA was substituted by BRACO19
DOI: 10.1039/D0CC06525K
(Figure S19) and thus it caused a significant fluorescence signal
reduction. These results may further support that the probe
binds to G4-DNA in live cells.
Cytotoxicity of BZT-Indolium against a number of cell lines
including cancer cells U87, HeLa, MCF7 and normal cells 16HBE
and HK2 were evaluated with MTT assays (Figure S20). From the
IC50 values (Table S5), the probe showed higher toxicity against
cancer cells U87, HeLa and MCF7 than the normal cells 16HBE
and HK2. The result is reasonable because c-MYC gene is
generally overexpressed in many cancer cells and thus the
probe may inhibit the expression of c-MYC gene in cells via the
stabilization of G4-structures.14-16 In addition, the less toxicity of
the probe against normal cells is an advantage for bio-
application in live cell experiments, particularly for those
require long-time video tracking of the bio-function and/or
dynamics of c-MYC promoter G4-DNA in live cells. More
importantly, BZT-Indolium bearing a small-sized and simple
molecular scaffold may give meaningful insights for the design
of cancer drugs targeting G4-structures of c-MYC oncogenes.
In conclusion, a small-sized G4-fluorescent probe was
demonstrated to be target-specific towards c-MYC promoter
G4-DNA with high photostability. The NMR titration and G4-
sequence mutation study showed that the probe may probably
interact with the loops of the G4-structure. The intracellular
competition and colocalization experiments in live cells showed
that the binding target of the probe in live cells presumably is a
G4-structure. The probe was found able to reveal the c-MYC G4-
DNA formation in live cells, stabilize the structures, inhibit the
amplification of the G4-sequence and down-regulate oncogene
c-MYC transcription human cancer cells.
Figure 5. Intracellular competition experiments in live U87 cells
and images were taken after incubated for 15 min: (A) 5 μM
probe; (B) 5 μM probe and then 5 μM BRACO19; (C) 5 μM probe
and then 10 μM BRACO19.
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This work was supported by the National Nature Science
Foundation of China (81473082, 22077020), Nature Science
Foundation of Guangdong Province, China (No.:
2017A030313078; 2017A030313071; 2019A1515011799), the
Department of Agriculture and Rural Affairs of Guangdong
Province, China (2018LM2175), and Jiangmen Program for
Innovative Research Team (No. 2018630100180019806).
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Conflicts of interest
There are no conflicts to declare.
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