Expression of NF-κB in hRPE cells stimulated by PDTC and IL-1β

Abstract

The constitutive expression of nuclear-factor-κB (NF-κB) in human pigment epithelial (hRPE) cells cultivatedin vitro and the possible changes when incubated with PDTC and IL-1β were investigated. The synchronized hRPE cellsin vitro were divided into two groups. In non-PDTC group, hRPE cells were exposed respectively to IL-1βand NS (for detecting the constitutive expressions of NF-κB in hRPE cells); In PDTC group, PDTC-pretreated hRPE cells were exposed respectively to IL-1βand NS (for detecting the constitutive expression of NF-κB in PDTC-pretreated hRPE cells). The expression of NF-κB in hRPE cells in two groups was detected by immunofluorescence stain and flow cytometry. The results showed that the constitutive expression of NF-κB in hRPE cellsin vitro was 8.05%, and increased to 30.26% by IL-1β. After PDTC pretreatment, the constitutive expression of NF-κB in hRPE cells was decreased to 3.74%, and 3.66% by IL-1βrespectively. It was concluded that the expressions of NF-κB in hRPE cells could be increased significantly by IL-1βand depressed effectively by PDTC. Also, PDTC could significantly inhibit the activation of NF-κB induced by IL-1β.

Keywords

pigment epithelial cell / nuclear-factor-κB / expression

References

[1] ZhengJ L, JinX M, ZhengH L, et al.. The cultane and evaluation of human RPE cells. Chinese J Practical Ophthalmology, 1999, 17(7): 407-407

[2] SenR, BaltimoreD. Multiple nuclear factors interact with the immunoglobulin enhancer sequences. Cell, 1986, 46(5): 705-705

[3] ZhaoJ, WuJ A. Expression of nudem transcription faton NF-kB in human Corntal fibroblasts. Chin J Ophthal mol, 1999, 35(1): 13-13

[4] WangX C, JobinC, AllenJ B, et al.. Suppression of NF-κB-dependent proinflammatory gene expression in human RPE cells by a proteasome inhibitor. Invest Ophthalmol Vis Sci, 1999, 40: 477-477

[5] SchreckR, RieberP, BaeuerleP A. Reactive oxygen intermediates as apparently widely used messengers in the activation of the NF-κB transcription factor and HIV-1. EMBO J, 1991, 10: 2247-2247

[6] BrennanP, O’NeillL A. 2-mercaptoethanol restores the ability of nuclear factor kappa B (NF kappa B) to bind DNA in nuclear extracts from interleukin 1-treated cells incubated with pyrollidine dithiocarbamate (PDTC). Evidence for oxidation of glutathione in the mechanism of inhibition of NF kappa B by PDTC. Biochem J, 1996, 320: 975-975

[7] ChenZ J, ParentL, ManiatisT. Site-specific phosphorylation of IkappaBalpha by a novel ubiquitination-dependent protein kinase activity. Cell, 1996, 84(6): 853-853

[8] TakadaY, AggarwalB B. Flavopiridol inhibits NF-kappaB activation induced by various carcinogens and inflammatory agents through inhibition of IkappaBalpha kinase and p65 phosphorylation: abrogation of cyclin D1, cyclooxygenase-2, and matrix metalloprotease-9. J Biol Chem, 2004, 279(6): 4750-4750