Abstract
The cytotoxic activity of curcumin towards CCRF-CEM human T-cell leukemia cells was measured by the MTT assay. Tumor cells were more sensitive to the cytotoxic activity of curcumin or curcumin-Cu (II) compared to normal cells, and the IC50 of curcumin towards CCRF-CEM cells was 8.68 µM, and that of curcumin-Cu (II) was 8.14 µM. The cell cycle distribution of curcumin-treated CCRF-CEM cells was analyzed by flow cytometry. DNA damage induced by oxidants such as curcumin-Cu (II) ions is considered as one of the main causes of cell inactivation. Therefore, we analyzed the effect of curcumin on DNA damage by agarose gel electrophoresis and atomic force microscopy (AFM). Gel electrophoresis analyses showed that curcumin or Cu (II) alone failed to cause DNA damage in pBR322 plasmid DNA as compared with the normal plasmid. However, DNA plasmids were mostly damaged after treatment with curcumin of different concentrations in the presence of Cu (II). Two forms were observed by means of AFM: closed circular plasmids and linear plasmids. DNA damage induced by a combination of curcumin and Cu (II) was also found by agarose gel electrophoresis, which was applied as control method to verify the results obtained by AFM.
Highlights
Curcumin is a natural pigment derived from turmeric (Curcuma longa L.), which has been known for its numerous pharmacological properties for many years [1,2,3]
The effects of curcumin and curcumin-Cu (II) on the viability of CCRF-CEM and Vero cells were determined by the MTT assay
The induction of DNA strand breakage by curcumin was assessed by measuring the conversion of supercoiled pBR322 plasmid DNA to open circular and linear forms or to further fragmentation by gel electrophoresis [27]. pBR322 DNA was treated with curcumin and/or Cu (II) in 10 mM Tris-HCl buffer at pH 7.4 and 37 oC for 1 h
Summary
Curcumin is a natural pigment derived from turmeric (Curcuma longa L.), which has been known for its numerous pharmacological properties for many years [1,2,3] It has the advantage of being nontoxic towards normal cells [4]. Curcumin has been considered as one of the most promising chemopreventive agents against a variety of human cancers [5] It may inhibit proliferation of cancer cells by cell cycle arrest and by inducing apoptosis [6,7]. Curcumin-induced DNA double-strand breaks have been determined by gel electrophoresis in previous study [12]. Our study showed that DNA double-strand breaks were induced after exposure to curcumin by applying AFM as novel detection technique for DNA damage.
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