G-quadruplex forming region within WT1 promoter is selectively targeted by daunorubicin and mitoxantrone: A possible mechanism for anti-leukemic effect of drugs.

Wilms tumor 1 (WT1) has long been overexpressed in acute myeloid leukemia and has a prognostic value in its diagnosis. Lately, the formation of G-quadruplexes in oncogenic promoters like (WT1) has been widely investigated since stabilization of these structures leads to transcriptional inhibition of the oncogene. Daunorubicin and mitoxantrone considered as crucial components of almost all standard acute myeloid leukemia induction regimens. Herein we have proposed a probable molecular mechanism of action through which the drugs may stabilize (WT1) promoter G-quadruplexes. Differential pulse voltammetry, circular dichroism, and polyacrylamide gel electrophoresis, electrophoretic mobility shifts assay, polymerase chain reaction (PCR) stop assays, and quantitative RT-PCR were performed in order to better understanding the nature of interactions between the drugs and G-quadruplexes. Data revealed that both drugs had potential to stabilize G-quadruplexes and down-regulate WT1 transcription but daunorubicin exposed more silencing impact. The results illustrated the therapeutic association of these two commercial FDA-approved drugs in (WT1) transcriptional down-regulation. Since (WT1) has known as a transcriptional regulator of at least 137 target genes, so the new data are significant for the development of new approaches to regulating WT1 and other target genes by employing special drugs in cancer treatment.

SiO2 nanoparticles modified CPE as a biosensor for determination of i-motif DNA/Tamoxifen interaction.

Cytosine-rich DNA sequences can form a highly ordered structure known as i-motif in slightly acidic solutions. The stability of the folded i-motif structure is a good strategy to inhibit the telomerase reaction in cancer cells. The electrochemical biosensor was prepared by modifying carbon paste electrode with SiO2 nanoparticles to investigate drugs which can stabilize this structure. Tamoxifen (Tam), an antiestrogen hormonal agent for treatment of breast cancer, was chosen as the model ligand and its interaction with i-motif structure was examined. The interaction between i-motif DNA and Tam was studied in PBS buffer and [Fe(CN)6](3-) through the cyclic voltammetry and square wave voltammetry methods. The oxidation peak of Tam, due to the i-motif DNA/Tam interaction, was observed after i-motif immobilized on the surface of the electrode. The i-motif formation was investigated by circular dichroism spectroscopy and the results showed that this structure can certainly be made with pH around 4.5, but its stability reduced by going to the more alkaline pH. The selectivity which was studied in the presence of complementary strand demonstrated that i-motif structure could be stabilized in acidic pH even in the presence of its complementary strand.

Voltammetric characterization of human telomeric G-quadruplex: A label free method for anticancer drug detection.

Human telomeric DNA typically consists of many tandem repeats of the guanine-rich sequences d (TTAGGG) termed an intermolecular Gquadruplex structure. This structure plays an important role in the protection, stabilization and replication of chromosome ends and so is an active target for therapeutic purposes. Recently ligands that are able to stabilize Gquadruplex structure, have received great attention because quadruplex-binding ligands have potential applications in cancer therapy. The screen-printed graphite electrode (SPE) was modified with synthesized SBA-N-propylpipyrazine-N-(2-mercaptopropane-1-one) (SBA-NPPNSH) mesoporous structure to investigate the Gquadruplex DNA (G4DNA) formation and stabilization. Differential pulse voltammetry was used to examine the stability and formation of G4DNA in various K(+) concentrations, under different pH conditions and also in the presence of positive and negative G4DNA-binding ligands. The stability effect of TMPyP4 as a positive G4DNA-binding ligand was examined in the presence of complementary G4DNA strands. This studying revealed that after adding K(+) or positive G4DNA-binding ligand a new peak observed in higher potential due to oxidation of guanine residuals in the Gquadruplex structure.

A switchable Gquadruplex device with the potential of a nanomachine for anticancer drug detection.

The unique ability of living systems to translate biochemical reactions into mechanical work has inspired the design of synthetic DNA motors which generate nanoscale motion via controllable conformational change. It is believable that G-quadruplex structures in certain regions of the genome may play a role in the poor maintenance of genomic stability, which is a characteristic of many types of cancers. In this regards, formation and stabilization of the quadruplex structures at the telomeric repeats is an effective way to hamper the telomere extension and blocking the elongation step. Here, we report a DNA machine for selective Gquadruplex-binding ligand recognition, based on a conformational change; the forces exerted by the precise DNA machine for Gquadruplex conformational change were probed via an electrical signal transducer electrochemically by differential pulse voltammetry and cyclic voltammetry. The proposed machine was prepared by modifying the screen-printed graphite electrode (SPE) with the synthesized SBA-N-propylpipyrazine-N-(2-mercaptopropane-1-one) (SBA@NPPNSH) mesoporous structures and Au nanoparticles (AuNPs). The thiolated functionalized groups of SBA@NPPNSH structures can help for preconcentration of the synthesize AuNPs on the surface. Then SH-G4DNA was linked to the modified electrode by an AuNPsS bond. The morphology of constructed machine was characterized by the Field emission scanning electron microscope (FESEM).

A human telomeric G-quadruplex-based electronic nanoswitch for the detection of anticancer drugs.

An electronic nanoswitch is described based on the conformational change of the DNA sequence in the presence of stabilizing ligands. The new electrochemical biosensor was prepared by modifying a screen-printed graphite electrode (SPE) with functionalized SiO2 nanoparticles [(SiO2-N-propylpiperazine-N-(2-mercaptopropane-1-one) (SiO2@NPPNSH)] and Au nanoparticles (AuNPs). These nanoparticles are able to immobilize thiolated G-quadruplex DNA structures (SH-G4DNA). The SH groups on the SiO2@NPPNSH nanoparticles provide a good platform for stabilizing AuNPs on the surface of the electrode. This is due to the fact that AuNPs are able to bind to the organic SH groups on the SiO2@NPPNSH. The SH-G4DNA binds to the modified electrode by a AuNPs-S bond. The structure of SiO2@NPPNSH was characterized by scanning electron microscopy (SEM), thermo-gravimetric analysis (TGA) and infrared (IR) spectroscopy. The morphology of the modified electrode was characterized by SEM. The interaction between G4DNA and the anticancer drug, Tamoxifen (Tam), was studied in Tris-HCl buffer and [Fe(CN)6](3-) using cyclic (CV) and square wave voltammetry (SWV). The G-quadruplex formation and the interaction mechanism were identified by circular dichroism (CD) measurements. The CV current was seen to decrease with increasing concentration of Tam due to interaction between G4DNA and Tam. This biosensor is a simple and useful tool for selecting G-quadruplex-binding ligands.