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: Applied Biosystems Life Technologies (Foster city, CA, USA) supplied the primers [1X; Assay IDs: Hs00819517_mH ( TRF1 ); Hs01554305_g1 ( TIN2 ); Hs00194619_m1 ( TRF2 ); Hs00368526_g1 ( TPP1 ); Hs00430292_m1 ( TRF2IP ); Hs00209984_m1 ( POT1 ); and Hs99999903_m1 ( β-actin )] and probe mixture, and AbGene Ltd. (Surrey, UK) supplied the ABsolute qPCR mix (1X) used to produce the 20-μl PCR reaction mixture.

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

Journal: BioMed Research International

Article Title: Time-Dependent Effects of POT1 Knockdown on Proliferation, Tumorigenicity, and HDACi Response of SK-OV3 Ovarian Cancer Cells

doi: 10.1155/2018/7184253

Figure Lengend Snippet: POT1 mRNA expression as determined by qRT-PCR (a) and POT1 protein expression as determined by western blot analysis (b) in shRNA-infected human ovarian cancer SK-OV3 cells. POT1-KD significantly decreased cell proliferation (c) and colony formation (d) in the early POT1-KD SK-OV3 cells. “Negative control” indicates negative control SK-OV3 cells that express a nonspecific shRNA, and “POT1-KD” represents POT1-knockdown SK-OV3 cells that express POT1-specific shRNA. (c) shows the cPDs of the SK-OV3 cells. The cPDs of the negative control cells are depicted with black dots and lines, while the cPDs of the immediate effect POT1-KD cells are depicted with black squares and lines. (d) shows images from the soft agar colony formation assay that used SK-OV3 cells. The dark dots represent the colonies. Images of the negative control cells and the immediate effect POT1-KD cells were captured via digital phase-contrast microscopy at 4x magnification; scale bar: 1000 μ m. ∗∗ P < 0.01, compared with negative control cells.

Article Snippet: We infected SK-OV3 cells with POT1 lenti-shRNA or nonspecific shRNA (Santa Cruz Biotechnology, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, shRNA, Infection, Negative Control, Knockdown, Soft Agar Assay, Microscopy

Journal: BioMed Research International

Article Title: Time-Dependent Effects of POT1 Knockdown on Proliferation, Tumorigenicity, and HDACi Response of SK-OV3 Ovarian Cancer Cells

doi: 10.1155/2018/7184253

Figure Lengend Snippet: Decreased c-Myc expression resulted in hTERT inhibition and reductions in telomerase activity and telomere length in the immediate effect POT1-KD cells. The mRNA expression levels of c-Myc and hTERT in the immediate effect POT1-KD cells and negative control cells (a); the relative telomerase activity levels in the immediate effect POT1-KD cells and negative control cells (b); and the relative telomere lengths in the immediate effect POT1-KD cells and negative control cells (c). ∗∗ P < 0.01, compared with negative control cells.

Article Snippet: We infected SK-OV3 cells with POT1 lenti-shRNA or nonspecific shRNA (Santa Cruz Biotechnology, USA).

Techniques: Expressing, Inhibition, Activity Assay, Negative Control

Journal: BioMed Research International

Article Title: Time-Dependent Effects of POT1 Knockdown on Proliferation, Tumorigenicity, and HDACi Response of SK-OV3 Ovarian Cancer Cells

doi: 10.1155/2018/7184253

Figure Lengend Snippet: Medium-term effect POT1-KD SK-OV3 cells displayed increased cell proliferation and more severe tumorigenicity than negative control cells. As shown in the cPD experiments, the cPDs of the negative control cells are depicted with black dots and lines, while the cPDs of the medium-term effect POT1-KD cells are depicted with black squares and lines (a). Images of the negative control and the medium-term effect POT1-KD cells were captured via digital phase-contrast microscopy at 4x magnification; scale bar: 1000 μ m. The dark dots represent the colonies in soft agar (b). ∗ P < 0.05, compared with negative control cells.

Article Snippet: We infected SK-OV3 cells with POT1 lenti-shRNA or nonspecific shRNA (Santa Cruz Biotechnology, USA).

Techniques: Negative Control, Microscopy

Journal: BioMed Research International

Article Title: Time-Dependent Effects of POT1 Knockdown on Proliferation, Tumorigenicity, and HDACi Response of SK-OV3 Ovarian Cancer Cells

doi: 10.1155/2018/7184253

Figure Lengend Snippet: POT1 mRNA expression was reduced, whereas c-Myc mRNA expression was increased in the medium-term effect POT1-KD cells, and as a result of the increase in c-Myc, hTERT mRNA expression was upregulated, telomerase activity was increased, and telomeres were elongated in the medium-term effect POT1-KD cells. The mRNA expression levels of POT1, c-Myc, and hTERT in the medium-term effect POT1-KD cells and negative control cells (a); the relative telomerase activity levels in the medium-term effect POT1-KD cells and negative control cells (b); and the relative telomere lengths in the medium-term effect POT1-KD cells and negative control cells (c). ∗∗ P < 0.01, compared with negative control cells.

Article Snippet: We infected SK-OV3 cells with POT1 lenti-shRNA or nonspecific shRNA (Santa Cruz Biotechnology, USA).

Techniques: Expressing, Activity Assay, Negative Control

Journal: BioMed Research International

Article Title: Time-Dependent Effects of POT1 Knockdown on Proliferation, Tumorigenicity, and HDACi Response of SK-OV3 Ovarian Cancer Cells

doi: 10.1155/2018/7184253

Figure Lengend Snippet: Knockdown of the POT1 gene induced a transient inhibitory effect on the JNJ-26481585 response of SK-OV3 cells, and c-Myc played an important role in the sensitization of SK-OV3 cells to the antitumor agent JNJ-26481585. The relative cell viability of the immediate effect POT1-KD cells and negative control cells (a) and the relative cell viability of the medium-term effect POT1-KD cells and negative control cells (b); the IC50 values of the immediate effect POT1-KD cells and negative control cells (c) and the IC50 values of the medium-term effect POT1-KD cells and negative control cells (d); the c-Myc mRNA expression levels in the immediate effect POT1-KD cells and negative control cells treated with JNJ-26481585 (e); the c-Myc mRNA expression levels in the medium-term effect POT1-KD cells and negative control cells treated with JNJ-26481585 (f); the c-Myc mRNA expression levels in the immediate effect POT1-KD cells and negative control cells before JNJ-26481585 treatment (g); the c-Myc mRNA expression levels in the medium-term effect POT1-KD cells and negative control cells before JNJ-26481585 treatment (h). ∗ P < 0.05, ∗∗ P < 0.01, and ∗∗∗ P > 0.05, compared with negative control cells.

Article Snippet: We infected SK-OV3 cells with POT1 lenti-shRNA or nonspecific shRNA (Santa Cruz Biotechnology, USA).

Techniques: Knockdown, Negative Control, Expressing