自噬是真核细胞中高度进化保守的生命过程，与细胞存活密切相关，将其作为抗肿瘤药物作用靶点是近年来肿瘤治疗热点研究方向之一。从生物毒素中提取抗肿瘤有效成分是药物研发的重要手段，是中医以毒攻毒理论治疗肿瘤的具体实践。多年来，我们对蜂毒素抗肝癌作用系列研究表明，其能多通路、多靶点抑制肝癌细胞增殖、诱导凋亡，近期又观察到蜂毒素能明显诱导人肝癌细胞自噬体形成，诱导自噬标记蛋白Beclin 1、LC3-Ⅱ高表达，初步证实自噬也是蜂毒素抑癌作用的靶点之一。本项目拟在前期研究的基础上，深入探讨蜂毒素诱导人肝癌细胞株HepG2、SMMC-7721自噬的效应和作用机制，基于Beclin 1复合物及自噬相关信号通路PI3K/Akt与RAS/MERK，明确蜂毒素诱导肝癌细胞株HepG2、SMMC-7721自噬的作用靶点，阐明诱导自噬在人肝癌细胞死亡中的作用，为蜂毒素的应用打下基础。

Autophagy is highly conservative process of life evolution, and it generally exists in eukaryotic cells. Autophagy is closely related to cell survival. Anticancer medicines which could target autophagy may become the new ones for therapeutic of cancer strategic treatment. It is a important drug development means to derive antitumor active ingredient from biological toxin for screening of natural medicine. This is also one of the ways of practice the theory of traditional Chinese medicine on cancer treatment. A systematic and depth study on melittin anti-liver cancer function was made in our department for many years. Our studies，both in vivo and in vitro，demonstrated that melittin could multi-channeling、multi-targeting inhibit HCC cell line on proliferation and induce apoptosis. By electron microscopy and confocal laser immunofluorescence on our re-preliminary experiments, we had observed that melittin could induce autophagy in human HCC cell line, and it could also induce autophagy marker protein Beclin 1, LC3-Ⅱoverexpression. All the initially results confirmed that autophagy is also one of the tumor suppressor multi-targets of melittin. Thus,in this project,we intends to observe melittin-induced autophagyin human hepatoma cell HepG2 and SMMC-7721 and analyse the mechanisms of autophagy in cell death, and we also focus on the autophagy-related signaling pathway AMPK, PI3K / Akt and RAS / MERK to explore targets melittin on human hepatocellular carcinoma HepG2, SMMC-7721 autophagic cell death. All the above experimental researches will help us do development and utilization of melittin.