Curcumin induces the expression of NF-κB and Bcl-2/Bax in human renal cell carcinoma cell line ACHN

Abstract

Objective To explore the in vitro effects of curcumin on the proliferation and apoptosis of the human renal cell carcinoma cell line ACHN, and to investigate its mechanisms of action. Methods The human renal cell carcinoma cell line ACHN was treated with different concentrations of curcumin for 24 h. The MTT assay was used to evaluate the cytotoxic effects of curcumin and flow cytometry was utilized to observe and detect the apoptosis of ACHN cells induced by curcumin. The expression levels of Bcl-2, Bax and NF-κBP65 mRNA were evaluated by Reverse Transcription-Polymerase Chain Reaction (RT-PCR), while the expression of Bcl-2, Bax, NF-κBP65 and IκB proteins was evaluated by Western blot. Results The concentrations of curcumin used significantly inhibited the proliferation of ACHN human renal cell carcinoma cells in vitro in a dose and time-dependent manner ( F time = 5.55, P < 0.05; F dose=110.05, P < 0.05). Obvious apoptosis of cells treated with different concentrations of curcumin could be observed by FCM. Compared with the control group, the apoptosis rates of curcumin-treated cells were markedly increased ( F=96.35, P < 0.05). Lower dose of curcumin significantly induced the apoptosis of ACHN cells. With intervention of different concentrations of curcumin (0, 10, 20 and 40 μmol/L) for 24 h, the expression levels of Bcl-2 and NF-κBP65 mRNA in ACHN cells were decreased while the expression level of Bax mRNA was increased ( P < 0.05), and Bcl-2, and NF-κBP65 protein decreased, while Bax and IκB protein increased compared with those in the untreated group. Conclusion Curcumin inhibited proliferation and increased apoptosis of the human renal cell carcinoma cell line ACHN. These curcumin effects appear to involve up-regulating IκB, down-regulating NF-κB, and regulating the expression of the apoptosis genes Bcl-2/Bax.