Raloxifene Increases Proliferation and Up-regulates Telomerase Activity in Human Umbilical Vein Endothelial Cells

Masakazu Doshida,Masahide Ohmichi,Seiji Tsutsumi,Jun Kawagoe,Toshifumi Takahashi,Botao Du,Akiko Mori-Abe,Tsuyoshi Ohta,Maki Saitoh-Sekiguchi,Kazuhiro Takahashi,Hirohisa Kurachi

doi:10.1074/jbc.m513251200

Masakazu Doshida, Masahide Ohmichi + Show 9 more

Open Access

https://doi.org/10.1074/jbc.m513251200

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Abstract

Vascular endothelial senescence is involved in human atherosclerosis. Telomerase activity is known to be critical in cellular senescence and its level is modulated by regulation of telomerase catalytic subunit (telomerase reverse transcriptase (TERT)) at both the transcriptional and post-transcriptional levels. Since the cardioprotective effect of estrogen itself has not been ruled out, we examined that of raloxifene, which has been classified as a selective estrogen receptor modulator, on the proliferation and telomerase activity of human umbilical vein endothelial cells (HUVECs). Raloxifene, like estrogen, clearly induced the telomerase activity and human TERT (hTERT) expression via estrogen receptor (ER) alpha and ERbeta. Treatment with raloxifene for 5 days significantly induced cell growth, and either cotreatment with a telomerase inhibitor, 3'-azido-3'-deoxythymidine, or transfection with hTERT-specific small interfering RNA significantly attenuated the raloxifene-induced cell growth. Raloxifene also induced the phosphorylation of Akt, and pretreatment with a phosphatidylinositol 3-kinase inhibitor, LY294002, significantly attenuated the raloxifene-induced telomerase activity. In addition, raloxifene induced both the phosphorylation of hTERT and IkappaB. Moreover, cotreatment with an IkappaBalpha phosphorylation inhibitor, BAY-11-7082, or a specific NFkappaB nuclear translocation inhibitor, SN50, significantly attenuated the raloxifene-induced telomerase activity and the association of NFkappaB with hTERT. These results show that raloxifene induced the up-regulation of telomerase activity not only by the transcriptional regulation of hTERT but also by post-translational regulation of the phosphorylation of Akt and hTERT and the association of hTERT with NFkappaB in HUVECs. Thus, the up-regulation of telomerase activity in vascular endothelial cells might be one mechanism contributing to the potential atheroprotective effect of raloxifene.

Highlights

The risk of cardiovascular disease steeply increases after menopause
Since the cardioprotective effect of estrogen itself has not been ruled out, we examined that of raloxifene, which has been classified as a selective estrogen receptor modulator, on the proliferation and telomerase activity of human umbilical vein endothelial cells (HUVECs)
Significant differences are indicated by asterisks. **, p Ͻ 0.01

Summary

Introduction

The risk of cardiovascular disease steeply increases after menopause. Many epidemiological and basic studies have shown that estrogen has the significant function in the vasculature of preventing the primary development of cardiovascular disease in women [1,2]. B, cells treated with or without (0 h) vehicle, 10 nM raloxifene (Ral ), or, 10 nM estrogen (E2) for 24 h were harvested, and the telomerase activity in each preparation was detected by the TRAP assay.

Results

Conclusion