In Vitro Modulation of Telomerase Activity, Telomere Length and Cell Cycle in MKN45 Cells by Verbascoside

Abstract

Screening of natural products with anti-tumor activity as telomerase inhibitor is a new subject in the field of tumor therapy. Using telomerase PCR ELISA, telomere DNA hybridization and flow cytometry analysis, the effects of verbascoside, a phenylpropanoid glucoside extracted from Pedicularis striata Pall, on telomerase activity, telomere length and cell cycle of human gastric carcinoma cells MKN45 was examined in vitro. After being treated with a 50 % inhibition concentration of verbascoside (17.8 μg/ml), telomerase activity in the cells was significantly inhibited but not in the cellular supernatant, the average telomere length became remarkably short, and the sub-G0 /G1 peak and G2/M arrest were also displayed when compared to the control cells. These results suggest that verbascoside mediated-cell differentiation and apoptosis may be affected by telomere-telomerase-cell cycle dependent modulation. Thus, the antitumor mechanism of verbascoside is demonstrated once more by its inhibiting effect on telomerase activity in tumor cells, and the telomerase assay may provide a valuable screening method for antitumor activity of natural products.

Abbreviations

PCR:polymerase chain reaction

ELISA:enzyme-linked immunosorbent assay

DDW:disinfected distillation water

TRAP:telomere repeat amplification protocol

PPG:phenylpropanoid glycoside

RT:room temperature

OD:optical density

^telIC₅₀:50 % inhibition of the telomerase activity

TRF:telomeric restriction fragments

FCM:flow cytometry

NC:nitrocellulose

Abstract

Screening of natural products with anti-tumor activity as telomerase inhibitor is a new subject in the field of tumor therapy. Using telomerase PCR ELISA, telomere DNA hybridization and flow cytometry analysis, the effects of verbascoside, a phenylpropanoid glucoside extracted from Pedicularis striata Pall, on telomerase activity, telomere length and cell cycle of human gastric carcinoma cells MKN45 was examined in vitro. After being treated with a 50 % inhibition concentration of verbascoside (17.8 μg/ml), telomerase activity in the cells was significantly inhibited but not in the cellular supernatant, the average telomere length became remarkably short, and the sub-G0 /G1 peak and G2/M arrest were also displayed when compared to the control cells. These results suggest that verbascoside mediated-cell differentiation and apoptosis may be affected by telomere-telomerase-cell cycle dependent modulation. Thus, the antitumor mechanism of verbascoside is demonstrated once more by its inhibiting effect on telomerase activity in tumor cells, and the telomerase assay may provide a valuable screening method for antitumor activity of natural products.

Abbreviations

PCR:polymerase chain reaction

ELISA:enzyme-linked immunosorbent assay

DDW:disinfected distillation water

TRAP:telomere repeat amplification protocol

PPG:phenylpropanoid glycoside

RT:room temperature

OD:optical density

^telIC₅₀:50 % inhibition of the telomerase activity

TRF:telomeric restriction fragments

FCM:flow cytometry

NC:nitrocellulose

Key words

Verbascoside - Pedicularis striata - Scrophulariaceae - telomere - telomerase - cell cycle - human gastric carcinoma cell MKN45

Key words

Verbascoside - Pedicularis striata - Scrophulariaceae - telomere - telomerase - cell cycle - human gastric carcinoma cell MKN45

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