The effect of endotoxin on functional parameters of mammary CID-9 cells

B. Safieh-Garabedian, Ghassan Mouneimne, W. El-Jouni, M. Khattar, R. Talhouk

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

The effect of endotoxin on mammary CID-9 cells, which differentiate in culture and express β-casein, was investigated. Cells in culture supplemented with lactogenic hormones and dripped with EMS-Matrix (EMS-drip), were treated daily with endotoxin (0.5-500 μg/ml). Endotoxin at concentrations of less or equal to 10 μg/ml did not affect cell growth and viability up to 5 days post endotoxin treatment. Endotoxin (0.01-10 μg/ml) was added to the culture medium, upon confluence, and functional parameters were examined within 48 h post endotoxin treatment. Nuclear factor-κB (NF-κB) (p52) increased in nuclear extracts from endotoxin-stimulated cells within 1 h of treatment, while β-casein mRNA and protein expression decreased in a concentration-dependent manner at 24 and 48 h post treatment. Zymography showed that the 72 and 92 kDa gelatinase activity increased in cells at 24 and 48 h post endotoxin treatment at 10 and 50 μg/ml. At the latter concentration, the active form of 72 kDa gelatinase was induced at 48 h. Interleukin-6 and tumor necrosis factor-α levels increased at 1-3 h post endotoxin treatment and peaked at 6 h in cells on plastic and EHS-drip. Nerve growth factor (NGF) levels increased in control and endotoxin-treated cells in a time-dependent manner, and endotoxin increased NGF levels in culture at 6 and 9 h post endotoxin treatment. This study shows that endotoxin activated NF-κB, suppressed β-casein expression and upregulated gelatinases, cytokines and NGF. This model could be used to investigate the role of mammary cells in initiating and propagating inflammation and to test candidate molecules for potential anti-inflammatory properties.

Original languageEnglish (US)
Pages (from-to)397-406
Number of pages10
JournalReproduction
Volume127
Issue number3
StatePublished - Mar 2004
Externally publishedYes

Fingerprint

Endotoxins
Breast
Nerve Growth Factor
Caseins
Matrix Metalloproteinase 2
Gelatinases
Matrix Metalloproteinase 9
Plastics
Culture Media
Interleukin-6
Cell Survival
Anti-Inflammatory Agents
Cell Culture Techniques
Tumor Necrosis Factor-alpha
Hormones
Cytokines
Inflammation

ASJC Scopus subject areas

  • Obstetrics and Gynecology
  • Cell Biology
  • Endocrinology
  • Embryology

Cite this

Safieh-Garabedian, B., Mouneimne, G., El-Jouni, W., Khattar, M., & Talhouk, R. (2004). The effect of endotoxin on functional parameters of mammary CID-9 cells. Reproduction, 127(3), 397-406.

The effect of endotoxin on functional parameters of mammary CID-9 cells. / Safieh-Garabedian, B.; Mouneimne, Ghassan; El-Jouni, W.; Khattar, M.; Talhouk, R.

In: Reproduction, Vol. 127, No. 3, 03.2004, p. 397-406.

Research output: Contribution to journalArticle

Safieh-Garabedian, B, Mouneimne, G, El-Jouni, W, Khattar, M & Talhouk, R 2004, 'The effect of endotoxin on functional parameters of mammary CID-9 cells', Reproduction, vol. 127, no. 3, pp. 397-406.
Safieh-Garabedian B, Mouneimne G, El-Jouni W, Khattar M, Talhouk R. The effect of endotoxin on functional parameters of mammary CID-9 cells. Reproduction. 2004 Mar;127(3):397-406.
Safieh-Garabedian, B. ; Mouneimne, Ghassan ; El-Jouni, W. ; Khattar, M. ; Talhouk, R. / The effect of endotoxin on functional parameters of mammary CID-9 cells. In: Reproduction. 2004 ; Vol. 127, No. 3. pp. 397-406.
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