J Korean Assoc Oral Maxillofac Surg.  2004 Aug;30(4):323-330.

Hsp27 contributes to estrogen regulation of osteoblast apoptosis

• Jang HS
• Eune JJ
• Rim JS
• Kwon JJ
• Choi CM

Abstract

Estrogen may promote osteoblast/osteocyte viability by limiting apoptotic cell death. We hypothesize that hsp27 is an estrogen- regulated protein that can promote osteoblast viability by increasing osteoblast resistance to apoptosis. The purpose of this study was to determine the effect of estrogen treatment and heat shock on TNF alpha- induced apoptosis in the MC3T3-E1 cell line. Cells were treated with 0 - 100 nM 17betaestradiol (or ICI 182780) for 0 - 24 hours before heat shock. After recovery, apoptosis was induced by treatment with 0 - 10 ng/ml TNF alpha. Hsp levels were evaluated by Northern and Western analysis using hsp27, hsp47, hsp70c and hsp70i - specific reagents. Apoptosis was revealed by in situ labeling with Terminal Deoxyribonucleotide Transferase (TUNEL). A 5 - fold increase in hsp27 protein and mRNA was noted after 5 hours of treatment with 10 - 20 nM 17beta estradiol prior to heat shock. Increased abundance of hsp47, hsp70c or hsp70i was not observed. TUNEL indicated that estrogen treatment also reduced (50%) MC3T3-E1 cell susceptibility to TNF alpha-induced apoptosis. Treatment with hsp27-specific antisense oligonucleotides prevented hsp27 protein expression and abolished the protective effects of heat shock and estrogen treatment on TNF alpha-induced apoptosis. Hsp27 is a determinant of osteoblast apoptosis, and estrogen treatment increases hsp27 levels in cultured osteoblastic cells. Hsp27 contributes to the control of osteoblast apoptosis and may be manipulated by estrogenic or alternative pathways for the improvement of bone mass.

Keyword

Heat Shock; Bone; Oligonucleotide; TNF-alpha; TUNEL

MeSH Terms

Apoptosis*
Cell Death
Cell Line
Estradiol
Estrogens*
Hot Temperature
HSP27 Heat-Shock Proteins
In Situ Nick-End Labeling
Indicators and Reagents
Oligonucleotides, Antisense
Osteoblasts*
RNA, Messenger
Shock
Transferases
Tumor Necrosis Factor-alpha
Estradiol
Estrogens
HSP27 Heat-Shock Proteins
Indicators and Reagents
Oligonucleotides, Antisense
RNA, Messenger
Transferases
Tumor Necrosis Factor-alpha