J Gynecol Oncol.  2010 Jun;21(2):119-124. 10.3802/jgo.2010.21.2.119.

Follicular stimulating hormone enhances Notch 1 expression in SK-OV-3 ovarian cancer cells

• Park YH
• Kim SJ
• Jeong BH
• Herzog TJ
• Wright J
• Kitajewski J
• Rhim CC
• Jang PR
• Kang JB
• Kim SJ

• KMID: 2009582
• DOI: http://doi.org/10.3802/jgo.2010.21.2.119

Abstract

OBJECTIVE
Notch is known as a transmembranous receptor family with four homologous forms - Notch 1, Notch 2, Notch 3, and Notch 4 and related to cell fate regulation and angiogenesis. The purpose is to investigate the effect of follicular stimulating hormone (FSH) on the Notch 1 expression and proliferation in ovarian cancer cells.
METHODS
Human ovarian cancer cell line, SK-OV-3 and FSH were used. XTT cell proliferation and cell migration assay were carried out with FSH 100 mIU/mL and Notch 1 siRNA. Western blots and reverse transcriptase-polymerase chain reactions (RT-PCR) were carried out to determine the expression level of the Notch 1 protein and mRNA with FSH treatment in 0, 1, 5, 10, 100, 200, 300 mIU/mL concentrations. Immunofluorescent (IF) stains were performed in SK-OV-3 cell cultures with FSH 100 mIU/mL. Student-t tests were used in statistical analyses.
RESULTS
The SK-OV-3 have Notch 1 receptors in their natural status. FSH stimulated SK-OV-3 cells in XTT cell proliferation and cell migration assays and notch 1 siRNA inhibited. The expression level of Notch 1 protein and mRNA were increased in a dose dependent pattern according to FSH concentrations compared to untreated cells. IF stains also showed brighter Notch1 expressions in the FSH treated cells compared to the control cells.
CONCLUSION
FSH enhances proliferation & migration and Notch 1 signaling in SK-OV-3 cells. The Notch signaling probably supports one of the cell proliferating mechanisms of FSH in ovarian cancer cells.

Keyword

Follicular stimulating hormone; Notch 1; Ovarian cancer

MeSH Terms

Blotting, Western
Cell Culture Techniques
Cell Line
Cell Migration Assays
Cell Proliferation
Coloring Agents
Humans
Ovarian Neoplasms
RNA, Messenger
RNA, Small Interfering
Coloring Agents
RNA, Messenger
RNA, Small Interfering

Figure

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  Fig. 1 The XTT assays of ovarian cancer cell line, SK-OV-3. (A) The cell proliferation effect of follicular stimulating hormone (FSH, 100 mIU/mL) in SK-OV-3 is significantly higher than the control (FSH 0 mIU/mL), 103 vs. 100 (p=0.0001, N=12). (B) Cell proliferations were inhibited significantly by Notch 1 siRNA in FSH 100 mIU/mL treated conditions 104.83 to 100.37 (p=0.009, N=8). *,†statistically significant, p<0.01.

• 

  Fig. 2 Cell migration assay by slit tests shows that follicular stimulating hormone (FSH) 100 mIU/ mL made more distant movement of cells dependent on time than the control (FSH 0 mIU/mL).

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  Fig. 3 The Notch 1 protein was examined by western blotting. HeLa and SK-OV-3 cells produce Notch 1 protein (300 KD) spontaneously without any drug treatment.

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  Fig. 4 RT-PCRs of Notch 1 mRNA in SK-OV-3 cells showing that follicular stimulating hormone (FSH) increases the gene expressions of Notch 1 in a dose dependent pattern. The lanes mean the concentration of the FSH 0, 1, 5, 10, 50, 100, 200, and 300 mIU/mL respectively. *statistically significant, p<0.05, †,‡statistically significant, p<0.01.

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  Fig. 5 The western blotting of Notch 1 protein (300 KD) in SK-OV-3 shows follicular stimulating hormonal (FSH) stimulatory effect. According to the FSH concentrations, the protein expressions are increasing. *statistically significant, p<0.05.

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  Fig. 6 The Notch1 immunofluorescent staining of SK-OV-3 cells with follicular stimulating hormone (100 mIU/ml). There are marked differences in fluorescence brightness. Negative control shows no signal (A), cells of right box (C) are much brighter than the cells of left one (B) (×200).

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