Effect Of Curcumin Analogue EF24on The Autophagy And Apoptosis In A549Cells And Its Toxicological Study

Curcumin is a traditional Chinese medicine, has a lot of pharmacologically active in vivo, in vitro, including anti-inflammatory, antioxidant, anti-aging, anti-virus, anti-tumor, and molecular targets at the intracellular.Curcumin that in organisms used efficiency is relatively low, it is difficult to dissolve in water and its easily oxidized characteristics, so in order to better improve its anti-tumor biological pharmacological activity of curcumin.The maternalappropriate transformation has become one of the direction of a new anticancer drug development.The biphenyl fluoroketone EF24[3,5-bis (2-flurobenzylidene) piperdin-4-one] is the transformation of the curcumin.Research display EF24is mainly concentrated in the reported anti-tumor promoting apoptosis, and its findings on the molecular target of the anti-tumor, but did not find EF24can cause tumor cell autophagy.The aim of our study is analyzed from the perspective of autophagy and apoptosis in the curcumin analogue EF24killing mechanism of tumor cell line A549(human lung adenocarcinoma cells), while selection of normal cell HEK293, used Caenorhabditis elegans as a model organism for preliminary evaluation of its toxicity.We choose different concentrations of EF24treatment of A549cells12h,24h,48h, MTT assay of cell viability, PI staining, GFP-LC3. Plasmid transfection, acridine orange staining, the protein of apoptosis, autophagy and AMPK/mTOR/S6K signaling pathway.Normal cells HEK293cell inhibitory rate of experimental and Caenorhabditis elegans movement-related indicators (head swinging frequency, bending frequency of the body), body length, reproductive indicators (number of offspring in the level of individual cell level and Caenorhabditis elegans,generation time) preliminary evaluation of the toxic effects of the EF24.Our study results showed that the EF24is able to inhibit the growth of A549cells, the better inhibitory effect of curcumin, and its inhibitory effect of showing the relationship of dose and time dependent.Autophagy and apoptosis detection in conditions of low concentration (4μmol/L8μmol/L), the cell can produce autophagy.Conditions of high concentration (EF24deal with A549cells24h16μmol/L) cells to apoptosis.EF24in A549cells by inhibition at the G2/M phase, and thus inhibit tumor cell growth.Since the autophagy inhibitor wortmannin join in. by acridine orange staining, the apoptotic protein caspase-3of from autophagy marker protein LC3detection and cell survival autophagy play in the process of apoptosis facilitating role.In the detection of pathway proteins, As EF24increase of the concentration, the increased level of AMPK phosphorylation and mTOR of S6K phosphorylation levels decreased.May be of AMPK-mTOR-S6K pathway to inhibit the growth of A549cells.In normal HEK293cells, the toxic effects of EF24is greater than curcumin, and less than cisplatin. And as EF24compounds concentration increased, in HEK293survival rate gradually decreased, its ability to kill normal cells than curcumin and less than cisplatin. Worm test, can be drawn below64μmol/L concentrations, did not produce toxic side effects, and256μmol/L, EF24nematode movement and reproductive impact and the EF24as analogues of curcumin than turmeric the toxicity of the factors to be rather less than cisplatin.Comprehensive evaluation EF24pharmacodynamic effect than curcumin side effects than cisplatin, applied to the clinical setting is a potential anticancer drugs.