Journal: Oncology Letters

Article Title: Trastuzumab increases the sensitivity of HER2-amplified human gastric cancer cells to oxaliplatin and cisplatin by affecting the expression of telomere-associated proteins

doi: 10.3892/ol.2014.2793

Figure Lengend Snippet: Effect of trastuzumab administration on the expression of telomere-associated genes and proteins in NCI-N87 cells. (A) Reverse transcription-quantitative polymerase chain reaction was used to determine the levels of TPP1 , POT1 , TRF1 , TRF2 , TERF2IP and TINF2 mRNA expression in NCI-N87 cells treated with trastuzumab and cisplatin (alone or in combination). Error bars indicate the standard deviation from the mean. * P<0.05, vs. control cells. (B) Western blot analysis determined that the protein expression levels of TPP1, POT1 and TRF2 in NCI-N87 cells were significantly inhibited by various concentrations of trastuzumab alone and low-dose trastuzumab in combination with cisplatin. TPP1 (formerly known as TINT1, PTOP and PIP); POT1, protection of telomere 1; TRF2, telomeric repeat binding factor 2; Control, untreated NCI-N87 cells.

Article Snippet: Following determination of protein concentration using a bicinchoninic acid protein assay kit (Pierce Biotechnology, Inc.), an aliquot of lysate containing 50 μg of each protein was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to polyvinylidene fluoride membranes, blocked with blocking buffer (PBS Tween-20 containing 5% non-fat milk) for 2 h at room temperature and incubated overnight at 4°C with the following specific primary antibodies: monoclonal mouse anti-human TPP1 (ACD; cat. no. TA504406), monoclonal rabbit anti-human POT1 (cat. no. TA310771), polyclonal rabbit anti-human TRF1 (cat. no. TA322887), rabbit anti-human monoclonal TRF2 (cat. no. TA307200), rabbit anti-human polyclonal TIN2 (cat. no. TA315321), rabbit anti-human polyclonal TRF2IP (cat. no. TA324532 ) and rabbit anti-human polyclonal GAPDH (cat. no. TA308884) (1:10,000, all primary antibodies; OriGene Technologies, Inc., Rockville, MD, USA).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Standard Deviation, Control, Western Blot, Binding Assay

Journal: Oncology Letters

Article Title: Trastuzumab increases the sensitivity of HER2-amplified human gastric cancer cells to oxaliplatin and cisplatin by affecting the expression of telomere-associated proteins

doi: 10.3892/ol.2014.2793

Figure Lengend Snippet: Effect of trastuzumab administration on the expression of telomere-associated genes and proteins in NCI-N87 cells. (A) Reverse transcription-quantitative polymerase chain reaction was used to determine the levels of TPP1 , POT1 , TRF1 , TRF2 , TERF2IP and TINF2 mRNA expression in NCI-N87 cells treated with trastuzumab and cisplatin (alone or in combination). Error bars indicate the standard deviation from the mean. * P<0.05, vs. control cells. (B) Western blot analysis determined that the protein expression levels of TPP1, POT1 and TRF2 in NCI-N87 cells were significantly inhibited by various concentrations of trastuzumab alone and low-dose trastuzumab in combination with cisplatin. TPP1 (formerly known as TINT1, PTOP and PIP); POT1, protection of telomere 1; TRF2, telomeric repeat binding factor 2; Control, untreated NCI-N87 cells.

Article Snippet: Following determination of protein concentration using a bicinchoninic acid protein assay kit (Pierce Biotechnology, Inc.), an aliquot of lysate containing 50 μg of each protein was subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis, transferred to polyvinylidene fluoride membranes, blocked with blocking buffer (PBS Tween-20 containing 5% non-fat milk) for 2 h at room temperature and incubated overnight at 4°C with the following specific primary antibodies: monoclonal mouse anti-human TPP1 (ACD; cat. no. TA504406), monoclonal rabbit anti-human POT1 (cat. no. TA310771), polyclonal rabbit anti-human TRF1 (cat. no. TA322887), rabbit anti-human monoclonal TRF2 (cat. no. TA307200), rabbit anti-human polyclonal TIN2 (cat. no. TA315321), rabbit anti-human polyclonal TRF2IP (cat. no. TA324532 ) and rabbit anti-human polyclonal GAPDH (cat. no. TA308884) (1:10,000, all primary antibodies; OriGene Technologies, Inc., Rockville, MD, USA).

Techniques: Expressing, Reverse Transcription, Real-time Polymerase Chain Reaction, Standard Deviation, Control, Western Blot, Binding Assay