In vitro effects of 2-methoxyestradiol on cell numbers, morphology, cell cycle progression, and apoptosis induction in oesophageal carcinoma cells

Abstract

The influence of 2-methoxyestradiol (2-ME) was investigated on cell numbers, morphology, cell cycle progression, and apoptosis induction in an oesophageal carcinoma cell line (WHCO3). Dose-dependent studies (1 x 10(-9)M-1 x 10(-6)M) revealed that 2-ME significantly reduced cell numbers to 60% in WHCO3 after 72 h of exposure at a concentration of 1 x 10(-6)M compared to vehicle-treated cells. Morphological studies entailing light-, fluorescent-, as well as transmission electron microscopy (TEM) confirmed 2-ME's antimitotic effects. These results indicated hallmarks of apoptosis including cell shrinkage, hypercondensation of chromatin, cell membrane blebbing, and apoptotic bodies in treated cells. Flow cytometric analyses demonstrated an increase in the G(2)/M-phase after 2-ME exposure; thus preventing cells from proceeding through the cell cycle. beta-tubulin immunofluorescence revealed that 2-ME caused spindle disruption. In addition, increased expression of death receptor 5 protein was observed further supporting the proposed mechanism of apoptosis induction via the extrinsic pathway in 2-ME-exposed oesophageal carcinoma cells.