摘要
目的研究姜黄素素诱导人宫颈癌细胞自噬的机制及对细胞功能的影响。方法采用10,20,和30μmol/L姜黄素处理宫颈癌SiHa细胞12h,随后采用Western-blot检测PI3K/Akt/mTOR信号通路中Akt和mTOR的磷酸化水平。同时采用自噬抑制剂3-MA处理细胞,采用MTT法观察处理前后SiHa细胞活性变化情况;ELISA检测IFN-γ的分泌水平,Western blot检测凋亡相关基因p53,p21和p27表达变化。结果姜黄素能以剂量依赖性方式抑制Akt和mTOR磷酸化。同时,姜黄素也能抑制SiHa细胞增殖并诱导其分泌IFN-γ。当同时给予3-MA处理后,可明显逆转姜黄素对SiHa生长增殖的抑制作用,同时也能消除姜黄素诱导其分泌IFN-γ。此外,姜黄素也能上调细胞内p53,p21和p27的表达,抑制自噬后,p53,p21和p27表达明显减少。结论姜黄素可通过抑制PI3K/Akt/mTOR活性诱导SiHa细胞自噬,最终抑制其增殖并诱导其分泌IFN-γ及表达p53,p21和p27。
Objective This study was designate to investigate the mechanism of Curcumin in the induction of authphagy and cellular function in human cervical cancer cells. Methods The human cervical cancer cell line SiHa was cultured and stimulated with 10,20,and 30 μmol/L of Curcumin for 12 h,the phosphorylated Akt and mTOR was detected by Western blot. Then cells were incubated with the autophagy inhibitor 3-MA,the cell activity was measured by MTT, the secretion of IFN-γ was detected by ELISA,and expression of the apoptosis associated gene p53,p21 and p27 were dectected by western blot. Results Curcumin could inhibit SiHa cells phosphorylation of Akt and roTOR. In addition, eurcumin could also inhibit SiHa cells proliferation and secretion of IFN-γ. Pretrement of 3-MA could significantly abrogate Curcumin-induced growth inhibition and IFN-7 production. Furthermore,Curcumin could also up-regulate Si- Ha cells expression of p53,p21 and p27,and this function could he reversed by 3-MA. Conclusion Curcumin could inhibit PI3K/Akt/mTOR activation to induce SiHa cells autophagy,and eventually decrease the growth activity and pro- mote the secretion of INF-γand expression of p53, p21 and p27.
出处
《中国实验诊断学》
2017年第5期897-900,共4页
Chinese Journal of Laboratory Diagnosis
基金
2015年深圳市科技研发资金基础研究资助项目(JCYJ20150331091010278)
