Curcumin inhibits AP-2γ-induced apoptosis in the human malignant testicular germ cells in vitro  

作　　者：Chang Zhou Xiao-meng Zhao Xiao-feng Li Cheng Wang Xiao-ting Zhang Xi-zhi Liu Xiao-feng Ding Shuang-lin Xiang Jian Zhang 

机构地区：[1]Department of Biochemistry and Molecular Biology, Key Laboratory of Protein Chemistry and Developmental Biology, Ministry of Education of China, College of Life Science, Hunan Normal University, Changsha 410081, China

出　　处：《Acta Pharmacologica Sinica》2013年第9期1192-1200,共9页中国药理学报（英文版）

基　　金：This work was supported by the National Natural Science Foundation of China （Grant Nos 81071696 and 31071150）; Project of Hunan Provincial Masters Innovation Fund （CX2012B227） and the Project of Chang Sha Science and Tech- nology Plan （K1109006-31, K1207014-31）.

摘　　要：Aim： To investigate the effects of curcumin on proliferation and apoptosis in testicular cancer cells in vitro and to investigate its molecular mechanisms of action.Methods： NTera-2 human malignant testicular germ cell line and F9 mouse teratocarcinoma stem cell line were used. The anti-proliferative effect was examined using MTT and colony formation assays. Hoechst 33258 staining, TUNEL and Annexin V-FITC/PI staining assays were used to analyze cell apoptosis. Protein expression was examined with Western blot analysis and immunocytochemical staining.Results： Curcumin （5, 10 and 15 μmol/L） inhibited the viability of NTera-2 cells in dose- and time-dependent manners. Curcumin significantly inhibited the colony formation in both NTera-2 and F9 cells. Curcumin dose-dependently induced apoptosis of NTera-2 cells by reducing FasL expression and Bcl-2-to-Bax ratio, and activating caspase-9, -8 and -3. Furthermore, curcumin dose-dependently reduced the expression of AP transcription factor AP-2γ in NTera-2 cells, whereas the pretreatment with the proteasome inhibitor MG132 blocked both the curcumin-induced reduction of AP-2γ and antiproliferative effect. Curcumin inhibited ErbB2 expression, and decreased the phosphorylation of Akt and ERK in NTera-2 cells.Conclusion： Curcumin induces apoptosis and inhibits proliferation in NTera-2 cells via the inhibition of AP-2γ-mediated downstream cell survival signaling pathways.Aim： To investigate the effects of curcumin on proliferation and apoptosis in testicular cancer cells in vitro and to investigate its molecular mechanisms of action.Methods： NTera-2 human malignant testicular germ cell line and F9 mouse teratocarcinoma stem cell line were used. The anti-proliferative effect was examined using MTT and colony formation assays. Hoechst 33258 staining, TUNEL and Annexin V-FITC/PI staining assays were used to analyze cell apoptosis. Protein expression was examined with Western blot analysis and immunocytochemical staining.Results： Curcumin （5, 10 and 15 μmol/L） inhibited the viability of NTera-2 cells in dose- and time-dependent manners. Curcumin significantly inhibited the colony formation in both NTera-2 and F9 cells. Curcumin dose-dependently induced apoptosis of NTera-2 cells by reducing FasL expression and Bcl-2-to-Bax ratio, and activating caspase-9, -8 and -3. Furthermore, curcumin dose-dependently reduced the expression of AP transcription factor AP-2γ in NTera-2 cells, whereas the pretreatment with the proteasome inhibitor MG132 blocked both the curcumin-induced reduction of AP-2γ and antiproliferative effect. Curcumin inhibited ErbB2 expression, and decreased the phosphorylation of Akt and ERK in NTera-2 cells.Conclusion： Curcumin induces apoptosis and inhibits proliferation in NTera-2 cells via the inhibition of AP-2γ-mediated downstream cell survival signaling pathways.

关 键 词：anticancer drugs CURCUMIN testicular germ cell tumors teratocarcinoma apoptosis AP-2γ MG132 ERBB2 Akt ERK 

分 类 号：Q255[生物学—细胞生物学] Q813