Journal: International Journal of Molecular Sciences

Article Title: Theaflavin-3,3′-Digallate Enhances the Inhibitory Effect of Cisplatin by Regulating the Copper Transporter 1 and Glutathione in Human Ovarian Cancer Cells

doi: 10.3390/ijms19010117

Figure Lengend Snippet: Treatment with TF3 potentiated inhibitory effect of cisplatin against ovarian cancer A2780/CP70 and OVCAR3 cells via upregulating CTR1 protein expression. ( A ) The effect of TF3 at the designated concentrations on the protein levels of MRP2, ATP7A, ATP7B and CTR1 in ovarian cancer cells; ( B ) treatment with 7.5 μM TF3 upregulated CTR1 protein levels in ovarian cancer cells pretreated with 7.5 μM cisplatin; ( C ) transfection with CTR1 siRNA decreased CTR1 protein levels in ovarian cancer cells; ( D ) transfection with CTR1 siRNA enhanced the resistance of ovarian cancer cells to 7.5 μM cisplatin. Results are expressed as mean ± SD from three independent experiments. Significant differences among different treatments are marked with different letters ( p < 0.05), * ( p < 0.05) and ** ( p < 0.01).

Article Snippet: CTR1 siRNA and control siRNA were purchased from Santa Cruz Biotechnology (Danvers, MA, USA).

Techniques: Expressing, Transfection

Journal: PLoS ONE

Article Title: EGCG Enhances Cisplatin Sensitivity by Regulating Expression of the Copper and Cisplatin Influx Transporter CTR1 in Ovary Cancer

doi: 10.1371/journal.pone.0125402

Figure Lengend Snippet: (A) Effect of EGCG on mRNA expression of CTR1, CTR2, ATP7A and ATP7B. After OVCAR3 cells were treated with indicated concentrations of EGCG for 24h, RT-PCR and qPCR were carried out to measure the CTR1 mRNA expression. (B and C) Effect of EGCG on CTR1 protein expression in OVCAR3 and SKOV3 cells. The cells were treated with 10 μM EGCG for 24 h, then followed by western blot analysis. The bands were quantified with Image J software. (* P <0.05, ** P <0.01)

Article Snippet: Human CTR1 siRNA or siRNA control (RiboBio, Guangzhou, China) were transfected into OVCAR3 and HEK-293T cells in a 96-well plate using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA), following the manufacturer’s instructions.Si-h-CTR1 sequences are listed below: si-h-CTR1 001: 5’ GAUCAAUACAGCUGGAGAA dTdT 3’ (forward) 3’dTdT CUAGUUAUGUCGACCUCUU 5’ (reverse); si-h-CTR1 002: 5’GGAAGAAGGCAGUGGUAGU dTdT 3’ (forward) 3’ dTdT CCUUCUUCCGUCACCAUCA 5’ (reverse); si-h-CTR1 003: 5’ CUACUUUGGCUUUAAGAAU dTdT 3’ (forward) 3’dTdT GAUGAAACCGAAAUUCUUA 5’ (reverse).

Techniques: Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Software

Journal: PLoS ONE

Article Title: EGCG Enhances Cisplatin Sensitivity by Regulating Expression of the Copper and Cisplatin Influx Transporter CTR1 in Ovary Cancer

doi: 10.1371/journal.pone.0125402

Figure Lengend Snippet: (A) Effect of knock-down of CTR1 on the sensitivity of ovary cancer cells to cDDP. OVCAR3 cells were transfected with three company sythetic si-RNAs or siRNA control and the western blost analysis showed si-RNA 3 exhibiting the best effect. After transfected with si-RNA3 or siRNA control, OVCAR3 and SKOV3 cells were treated with cDDP at various doses for 48 h and the cell survival fraction was detected by MTT assay. (B) Embryonic kidney HEK-293 cells were tranfected with human CTR1 si-RNA3 or siRNA control. Then the cells were exposed by indicated doses of cDDP for 48 h and the cell survival fraction was detected by MTT assay. (* P <0.05).

Article Snippet: Human CTR1 siRNA or siRNA control (RiboBio, Guangzhou, China) were transfected into OVCAR3 and HEK-293T cells in a 96-well plate using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA), following the manufacturer’s instructions.Si-h-CTR1 sequences are listed below: si-h-CTR1 001: 5’ GAUCAAUACAGCUGGAGAA dTdT 3’ (forward) 3’dTdT CUAGUUAUGUCGACCUCUU 5’ (reverse); si-h-CTR1 002: 5’GGAAGAAGGCAGUGGUAGU dTdT 3’ (forward) 3’ dTdT CCUUCUUCCGUCACCAUCA 5’ (reverse); si-h-CTR1 003: 5’ CUACUUUGGCUUUAAGAAU dTdT 3’ (forward) 3’dTdT GAUGAAACCGAAAUUCUUA 5’ (reverse).

Techniques: Knockdown, Transfection, Control, Western Blot, MTT Assay

Journal: PLoS ONE

Article Title: EGCG Enhances Cisplatin Sensitivity by Regulating Expression of the Copper and Cisplatin Influx Transporter CTR1 in Ovary Cancer

doi: 10.1371/journal.pone.0125402

Figure Lengend Snippet: (A, B) The effect of cDDP on the expression of CTR1 in OVCAR3 and SKOV3 cells. The cells were treated with 10 μM cDDP for the indicated time and CTR1 protein expression were detected by western blot analysis. (C)The effect of MG132 on the degradation of CTR1. After OVCAR3 cells were pretreated with 5 μM MG132 for 10h, the cells were incubation with 10 μM cDDP for 14 h. Then followed by western blot analysis. (D, E) The effect of EGCG on cDDP-trigged decrease of CTR1. The OVCAR3 and SKOV3 cells were treated with/without 10μM EGCG in the presence/absence of 10μMcDDP for 24 h. CTR1 protein expression was detected. The bands were quantified with Image J software. (* P <0.05, ** P <0.01)

Article Snippet: Human CTR1 siRNA or siRNA control (RiboBio, Guangzhou, China) were transfected into OVCAR3 and HEK-293T cells in a 96-well plate using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA), following the manufacturer’s instructions.Si-h-CTR1 sequences are listed below: si-h-CTR1 001: 5’ GAUCAAUACAGCUGGAGAA dTdT 3’ (forward) 3’dTdT CUAGUUAUGUCGACCUCUU 5’ (reverse); si-h-CTR1 002: 5’GGAAGAAGGCAGUGGUAGU dTdT 3’ (forward) 3’ dTdT CCUUCUUCCGUCACCAUCA 5’ (reverse); si-h-CTR1 003: 5’ CUACUUUGGCUUUAAGAAU dTdT 3’ (forward) 3’dTdT GAUGAAACCGAAAUUCUUA 5’ (reverse).

Techniques: Expressing, Western Blot, Incubation, Software

Journal: PLoS ONE

Article Title: EGCG Enhances Cisplatin Sensitivity by Regulating Expression of the Copper and Cisplatin Influx Transporter CTR1 in Ovary Cancer

doi: 10.1371/journal.pone.0125402

Figure Lengend Snippet: Four groups (control, EGCG, cDDP and EGCG+cDDP) were set up. Except there were 6 mice in control group, there were 8 mice for each of the other groups. The body weight (A) and the tumor size (A) were measured twice a week. (B) The mRNA expression of the CTR1 in tumor tissues was measured by RT-PCR and real qPCR. (C) The expression of CTR1 in tumor tissue was assessed by western blotting. (D) The expression of CTR1 in kidney tissue was measured by western blotting. The bands were quantified by Image J software. (*P<0.05, **P<0.01)

Article Snippet: Human CTR1 siRNA or siRNA control (RiboBio, Guangzhou, China) were transfected into OVCAR3 and HEK-293T cells in a 96-well plate using Lipofectamine 2000 reagent (Invitrogen, Carlsbad, CA, USA), following the manufacturer’s instructions.Si-h-CTR1 sequences are listed below: si-h-CTR1 001: 5’ GAUCAAUACAGCUGGAGAA dTdT 3’ (forward) 3’dTdT CUAGUUAUGUCGACCUCUU 5’ (reverse); si-h-CTR1 002: 5’GGAAGAAGGCAGUGGUAGU dTdT 3’ (forward) 3’ dTdT CCUUCUUCCGUCACCAUCA 5’ (reverse); si-h-CTR1 003: 5’ CUACUUUGGCUUUAAGAAU dTdT 3’ (forward) 3’dTdT GAUGAAACCGAAAUUCUUA 5’ (reverse).

Techniques: Control, Expressing, Reverse Transcription Polymerase Chain Reaction, Western Blot, Software