| Purpose: To explore whether cervical cancer cells co-express RANKL and RANK, and whether the RANKL/RANK system regulates the proliferation and apoptosis of cervical cancer cells and the possible mechanism as well as the relationship with IL-8.Methods: We evaluated the expression of RANKL/RANK in cervical cancer tissues and paracancerous cells by immunohistochemistry. Flow cytometry was used to detect the expression of RANKL and RANK in He La and Si Ha cells, CXCR1 and CXCR2(the receptors of IL-8) expression in the He La and Si Ha cells after treatment with anti-RANKL neutralizing(α-RANKL), along with the expression of proliferation- and apoptosis-related molecules in cervical cancer cells after treatment with α-RANKL or recombinant human IL-8protein(rh IL-8). We also used Enzyme-linked immunosorbent assay to test the secretion of soluble RANKL(s RANKL) from the He La and Si Ha cells and the secretion of IL-8 in the He La and Si Ha cells after treatment with α-RANKL. Besides, Brd U cell proliferation assay and Annexinv/PI apoptosis assay were performed to investigate the effects of the RANKL/RANK system and IL-8 on the abilities of the He La and Si Ha cells to proliferate and undergo apoptosis.Results: The cervical cancer tissues exhibited strong positive staining for RANKL and RANK compared to paracancerous cells. He La and Si Ha cells co-expressed member RANKL(m RANKL) and its receptor RANK, and also secrete s RANKL. Blocking RANKL/RANK interaction with α-RANKL or rh OPG led to a decrease of cell proliferation and a increase of apoptosis in He La and Si Ha cells. Besides, the decrease in cell proliferation induced by blocking RANKL/RANK with α-RANKL was more apparent in the group with high cell density. Further more, α-RANKL and rh OPG markedly downregulated Ki-67 and Bcl-2expression and upregulated Fas and Fas L expression in the He La and Si Ha cells. RANKL stimulated IL-8 secretion but decreased CXCR1 and CXCR2 expression in the He La and Si Ha cells. In addition, rh IL-8 reversed the effect of α-RANKL on proliferation- and apoptosis-related molecules, proliferation and apoptosis of He La and Si Ha cells.Conclusions: Cervical cancer cells highly co-express RANKL and RANK. Member RANKL/RANK interaction promotes the growth of cervical cancer cells by strengthening the crosstalk between cervical cancer cells and regulation of IL-8 secretion, and then stimulates the development of cervical cancer. |
