[Effects of PI3K/Akt/NF-κB signal pathway on FSH facilitation on cell proliferation and invasion by human epithelial ovarian cancer]

Chun-lin Xu, Xiao-lin Lu, Xiao-nan Yan, Hui-lan Wang, Su-qin Chen
Zhonghua Fu Chan Ke za Zhi 2012, 47 (2): 134-8

OBJECTIVE: To explore the effects of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt)/nuclear factor-kappa B (NF-κB) signal pathway on the process of follicle-stimulating hormone (FSH) facilitating cell proliferation and invasion in human epithelial ovarian cancer.

METHODS: Ovarian cancer cell lines SKOV3 and 3AO were cultured to exponential phase, then assigned to control group, FSH group, LY294002 group and FSH + LY294002 group, respectively. Cells were treated with different concentration of FSH and LY294002, respectively. The effects of FSH on cell proliferation were observed by methylthiazolyl tetrazolium (MTT). Morphological changes were observed by phase contrast microscope. The ability of cell invasion was investigated by transwell invasion assay. The expression of FSH receptor (FSHR), Akt1/2, phosphorylated-Akt (p-Akt) and NF-κB p65 protein were detected by western blot.

RESULTS: (1) FSH could promote the proliferation of SKOV3 and 3AO cells. When the cells were treated with 40 U/L FSH for 48 hours (SKOV3) and 24 hours (3AO), compared with those in control groups, they reached the highest proliferation rate (P < 0.05), respectively. (2) The morphology of SKOV3 and 3AO cells in four groups:in control group, SKOV3 cells were short spindle and 3AO cells were long spindle, the nuclei of them were both roundness or oval, the cytoplasm were bright. In FSH group, the cells changed to slightly longer or polygonal, they were full in shape, meanwhile, the cell intensity were higher than control group. In LY294002 group, some cells changed from spindle to round, and began to shrink. The cell intensity diminished. The morphology of FSH + LY294002 group was similar with control group, but the cell intensity was lower than that in FSH group. (3) The number of SKOV3 cell that passed through the membrane in control group, FSH group, LY294002 group and FSH + LY294002 group was (26 ± 6), (118 ± 19), (18 ± 5) and (38 ± 7), respectively. The number of 3AO cell was (19 ± 4), (134 ± 20), (12 ± 3) and (58 ± 11), respectively. The results showed that the number of cells in FSH group was significantly higher than that in control group (P < 0.05), while the number of cell in FSH + LY294002 group was significantly fewer than that in FSH group (P < 0.05). (4) There was no significant difference in the expression of FSHR and Akt1/2 between FSH group and control group (P > 0.05), but FSH increased the expression of p-Akt and the ratio of NF-κB p65 in the nucleus versus cytoplasm in SKOV3 and 3AO cells, there were significant differences compared with control group (P < 0.05). LY294002 reversed the effects of FSH on increasing the expression of p-Akt and the ratio of NF-κB p65 in the nucleus versus cytoplasm, there were significant differences among LY294002 group, FSH + LY294002 group and FSH group (P < 0.05).

CONCLUSION: The effects of FSH on proliferation and invasion of ovarian cancer cell lines SKOV3 and 3AO may be realized by regulating the activity of NF-κB in PI3K/Akt signal pathway.

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