Abstract
Lifeguard (LFG) is an anti-apoptotic protein that inhibits Fas-mediated death in tumour cells. However, the molecular function of human LFG in the carcinogenesis of human breast cells is uncertain. We studied the expression and function of endogenous LFG in four breast cancer cell lines (MCF-7, MDA-MB-231, T-47D and HS 578T), a human breast epithelial cell line (HS 578Bst), and in healthy and cancerous breast tissues. Molecular (Western blot and RT-PCR) and immunohistochemical techniques were used to investigate the LFG expression. To investigate the breast cancer cell proliferation in the presence of Fas, we performed fluorescent cell viability assays. The possible association of Fas with LFG was analyzed by immunofluorescence microscopy. In this paper, we provide convincing evidence that LFG is overexpressed in several human breast cancer cell lines. More importantly, we found that the LFG expression correlates with high tumour grades in primary breast tumours. Finally, we demonstrated that Fas sensitivity is reduced in breast cancer cell lines expressing LFG. Our results indicated that LFG is strongly expressed in breast cancer epithelial cells. Moreover, the overexpression of LFG correlated with tumour grade and reduced Fas sensitivity. Our findings support the idea that LFG may have a role in the downregulation of apoptosis in breast cancer cells.
Key words: Apoptosis, Lifeguard, Fas, Breast cancer
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Abbreviations used
- BI-1
Bax inhibitor-1
- DAPI
4’, 6’-diamidino-2-phenylindole
- EDTA
ethylenediaminetetraacetic acid
- EGF
epidermal growth factor
- FCS
fetal calf serum
- LFG
Lifeguard
- RT-PCR
reverse transcribed polymerase chain reaction
- PMSF
phenylmethyl sulfonyl fluoride
- PVDF
polyvinylidene fluoride
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