Journal: Biomedicines

Article Title: Structure of Some Green Tea Catechins and the Availability of Intracellular Copper Influence Their Ability to Cause Selective Oxidative DNA Damage in Malignant Cells

doi: 10.3390/biomedicines10030664

Figure Lengend Snippet: Elevated mRNA transcript levels of copper transporters Ctr1 and ATP7A in MCF-10A-Cu cells, relative to the parental MCF-10A cells, and the effect of EGCG. Total RNA was isolated by using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. A real-time PCR was used to quantify Ctr1 and ATP7A mRNA expression as described in the Materials and Methods section. Only MCF-10A-Cu (normal MCF-10A cells cultured in a medium containing 25 µM CuCl 2 ), with elevated mRNA expression of copper transporters, was subjected to treatment with EGCG (50 µM) to assess the effect of EGCG on mRNA expression. Values reported are ±S.E. of three independent experiments. * p value < 0.01 when compared to control.

Article Snippet: Sequences of primers for Ctr1 (forward: 5′-GCT GGA AGA AGG CAG TGG TA-3′; reverse: 5′-AAA GAG GAG CAA GAA GGG ATG-3′), ATP7A (forward: 5′-ACG AAT GAG CCG TTG GTA GTA-3′; reverse: 5′-CCT CCT TGT CTT GAA CTG GTG-3′) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (forward: 5′-TGG GTG TGA ACC ATG AGA AGT-3′; reverse: 5′-TGA GTC CTT CCA CGA TAC CAA-3′) were the same as reported earlier [ , ], and the amount of RNA was normalized for GAPDH expression. siRNA (small interfering RNA) transfection: siRNA transfections were performed, as described previously [ ]. siRNA specific to ctr1 was purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, Dallas, TX, USA).

Techniques: Isolation, Real-time Polymerase Chain Reaction, Expressing, Cell Culture, Control

Journal: Biomedicines

Article Title: Structure of Some Green Tea Catechins and the Availability of Intracellular Copper Influence Their Ability to Cause Selective Oxidative DNA Damage in Malignant Cells

doi: 10.3390/biomedicines10030664

Figure Lengend Snippet: Elevated mRNA transcript levels of copper transporters Ctr1 and ATP7A in MCF-10A-Cu cells, relative to the parental MCF-10A cells, and the effect of EGCG. Total RNA was isolated by using TRIzol reagent (Invitrogen, Carlsbad, CA, USA) according to the manufacturer’s instructions. Real-time PCR was used to quantify Ctr1 and ATP7A mRNA expression as described in the Materials and Methods section. Only MCF-10A-Cu (normal MCF-10A cells cultured in a medium containing 25 µM CuCl 2 ), with elevated mRNA expression of copper transporters, was subjected to treatment with EGCG (50 µM) to assess the effect of EGCG on mRNA expression. Values reported are ±S.E. of three independent experiments. * p value < 0.01 when compared to respective control.

Article Snippet: Sequences of primers for Ctr1 (forward: 5′-GCT GGA AGA AGG CAG TGG TA-3′; reverse: 5′-AAA GAG GAG CAA GAA GGG ATG-3′), ATP7A (forward: 5′-ACG AAT GAG CCG TTG GTA GTA-3′; reverse: 5′-CCT CCT TGT CTT GAA CTG GTG-3′) and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (forward: 5′-TGG GTG TGA ACC ATG AGA AGT-3′; reverse: 5′-TGA GTC CTT CCA CGA TAC CAA-3′) were the same as reported earlier [ , ], and the amount of RNA was normalized for GAPDH expression. siRNA (small interfering RNA) transfection: siRNA transfections were performed, as described previously [ ]. siRNA specific to ctr1 was purchased from Santa Cruz Biotechnology, Inc (Santa Cruz, Dallas, TX, USA).

Techniques: Isolation, Real-time Polymerase Chain Reaction, Expressing, Cell Culture, Control

Journal: Oncotarget

Article Title: Dextran-Catechin: An anticancer chemically-modified natural compound targeting copper that attenuates neuroblastoma growth

doi: 10.18632/oncotarget.10201

Figure Lengend Snippet: ( A ) Representative Western blot for CTR1 protein on whole-cell extracts from IMR-32, IMR-32-CisRes, BE(2)-C, and MRC-5 cells. GAPDH expression was used as a protein loading control. ( B ) Densitometry graph of Western blots showing higher expression of CTR1 in IMR-32 and BE(2)-C cells compared to IMR-32-CisRes and normal MRC-5 cells. Values are normalized to GAPDH protein expression and shown relative to CTR1 expression in IMR-32 cells (100%). ( C ) Intracellular copper levels are higher in IMR-32 and BE(2)-C cells compared to IMR-32-CisRes and MRC-5 cells. Columns , means of at least three independent experiments; Bars , SEM (* P < 0.05, ** P < 0.01).

Article Snippet: Twelve hours post-transfection with two different CTR1 specific siRNAs (siRNA A and siRNA B Origene, Rockville, MD, USA) or scrambled non-silencing siRNA, cells were treated with Dextran-Catechin for 24 hours.

Techniques: Western Blot, Expressing, Control

Journal: Oncotarget

Article Title: Dextran-Catechin: An anticancer chemically-modified natural compound targeting copper that attenuates neuroblastoma growth

doi: 10.18632/oncotarget.10201

Figure Lengend Snippet: ( A ) Cell death in IMR-32 and ( B ) BE(2)-C cells after knockdown of CTR1 and subsequent treatment with Dextran-Catechin for 24 hours. Columns , means of at least three independent experiments; Bars , SEM (*** p < 0.001, **** p < 0.0001).

Article Snippet: Twelve hours post-transfection with two different CTR1 specific siRNAs (siRNA A and siRNA B Origene, Rockville, MD, USA) or scrambled non-silencing siRNA, cells were treated with Dextran-Catechin for 24 hours.

Techniques: Knockdown