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. 2016 Mar 7:11:873-83.
doi: 10.2147/IJN.S100903. eCollection 2016.

Apoptosis inducing ability of silver decorated highly reduced graphene oxide nanocomposites in A549 lung cancer

Merajuddin Khan  1 , Mujeeb Khan  1 , Abdulhadi H Al-Marri  1 , Abdulrahman Al-Warthan  1 , Hamad Z Alkhathlan  1 , Mohammed Rafiq H Siddiqui  1 , Vadithe Lakshma Nayak  2 , Ahmed Kamal  2 , Syed F Adil  1
Affiliations

Apoptosis inducing ability of silver decorated highly reduced graphene oxide nanocomposites in A549 lung cancer

Merajuddin Khan et al. Int J Nanomedicine. .

Abstract

Recently, graphene and graphene-based materials have been increasingly used for various biological applications due to their extraordinary physicochemical properties. Here, we demonstrate the anticancer properties and apoptosis-inducing ability of silver doped highly reduced graphene oxide nanocomposites synthesized by employing green approach. These nano composites (PGE-HRG-Ag) were synthesized by using Pulicaria glutinosa extract (PGE) as a reducing agent and were evaluated for their anticancer properties against various human cancer cell lines with tamoxifen as the reference drug. A correlation between the amount of Ag nanoparticles on the surface of highly reduced graphene oxide (HRG) and the anticancer activity of nanocomposite was observed, wherein an increase in the concentration of Ag nanoparticles on the surface of HRG led to the enhanced anticancer activity of the nanocomposite. The nanocomposite PGE-HRG-Ag-2 exhibited more potent cytotoxicity than standard drug in A549 cells, a human lung cancer cell line. A detailed investigation was undertaken and Fluorescence activated cell sorting (FACS) analysis demonstrated that the nanocomposite PGE-HRG-Ag-2 showed G0/G1 phase cell cycle arrest and induced apoptosis in A549 cells. Studies such as, measurement of mitochondrial membrane potential, generation of reactive oxygen species (ROS) and Annexin V-FITC staining assay suggested that this compound induced apoptosis in human lung cancer cells.

Keywords: anticancer; apoptosis; graphene/silver nanocomposites; plant extract.

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Figures

Figure 1
Figure 1
Platinum (Pt)-containing anticancer drugs.
Figure 2
Figure 2
Graphical representation of the green synthesis of graphene/silver (PGE-HRG-Ag) nanocomposites using Pulicaria glutinosa plant extract and their anticancer activities against various human cancer cell lines. Abbreviations: T, temperature, t, time.
Figure 3
Figure 3
(A) UV-Vis absorption spectra of GRO, PGE-HRG, and PGE-HRG-Ag; (B) XRD spectra of GRO, PGE-HRG, and PGE-HRG-Ag nanocomposites. Abbreviation: GRO, graphene oxide.
Figure 4
Figure 4
Raman spectra of graphene/silver nanocomposites with 50 wt% (PGE-HRG-Ag-1, pink line) and 100 wt% (PGE-HRG-Ag-2, green line) of Ag NPs with respect to graphene and PGE-HRG without Ag NPs (blue line). Notes: Increasing the concentration of Ag NPs, the intensities of the Raman signals also increase, which not only confirms the binding of the Ag NPs on the surface of graphene but also demonstrates the increased density of the Ag NPs. Abbreviation: NPs, nanoparticles.
Figure 5
Figure 5
High resolution TEM images of (A) PGE-HRG-Ag-1 and (B) PGE-HRG-Ag-2 prepared using 0.5 and 1 mmol of AgNO3, respectively. Abbreviation: TEM, transmission electron microscopy.
Figure 6
Figure 6
FACS analysis in A549 human lung cancer cell line after treatment with PGE-HRG-Ag-2. Notes: (A) Untreated control cells (A549), (B) PGE-HRG-Ag-2 (5 μg), and (C) PGE-HRG-Ag-2 (10 μg). Abbreviation: FACS, fluorescence-activated cell sorting.
Figure 7
Figure 7
Drops in membrane potential (ΔΨm) were assessed by JC-1 staining of MCF-7 cells treated with compound PGE-HRG-Ag-2 and samples were then subjected to flow cytometry analysis on a FACScan (Becton Dickinson). Notes: (A) Untreated control cells (A549), (B) PGE-HRG-Ag-2 (5 μg), and (C) PGE-HRG-Ag-2 (10 μg). Abbreviations: FACS, fluorescence-activated cell sorting; ΔΨm, mitochondrial membrane potential; Q1-UL, quadrant 1-upper left; Q1-UR, quadrant 1-upper right; Q1-LL, quadrant 1-lower left; Q1-LR, quadrant 1-lower right.
Figure 8
Figure 8
The effect of PGE-HRG-Ag-2 on the ROS production in human lung cancer (A549). Notes: (A) Untreated control cells (A549), (B) PGE-HRG-Ag-2 (5 μg), and (C) PGE-HRG-Ag-2 (10 μg). Abbreviation: ROS, reactive oxygen species.
Figure 9
Figure 9
Annexin V-FITC staining. Notes: (A) Untreated control cells (A549), (B) PGE-HRG-Ag-2 (5 μg), and (C) PGE-HRG-Ag-2 (10 μg). Abbreviations: Q1-UL, quadrant 1-upper Left; Q1-UR, quadrant 1-upper right; Q1-LL, quadrant 1-lower left; Q1-LR, quadrant 1-lower right.
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