Role Of Autophagic Death In The Radiosensititly-induced By Aged Black Garlic Extract On Liver Cancer Huh7

Purpose: To analyze the role of autophagic death in the radiosensititly-induced by aged black garlic extract Huh7 from liver cancer and its possible mechanism. Methods: Choose Huh7 cell line as research object. Conventional culture of liver cancer Huh7 cells and classify them into four groups: (1) control group, (2) O.lg/ml, 0.3g/ml, 0.5g/ml, 0.7g/ml, 1.0g/ml concentration of aged black garlic extract group, (3)0Gy,2Gy,4Gy,6Gy,8Gy 60Coyray as radiation group,(4) High and Low concentration of aged black garlic extract plus radiation group. Inhibition rate of liver cancer was tested by MTT. Effect of colony formation was checked by flat plate colony forming. Flow Cytometry was used to checked changes in the cell cycle and apoptosis. Electron microscopy was used to detect autophagy. Beclinl and LC3,which was related to autophagy and the relationship between the concentration of aged black garlic extract were checked by Western blot. Results: 1.Certain concentration aged black garlic extract could had significantly inhibitory on proliferation of liver cancer Huh7, and relationshiped with concentration and acting time. Increase concentration of aged black garlic extract or acting time would reduce cell active. 2. Radiation group and aged black garlic extract plus radiation group cloning efficiency were 100%, 82.95%, 31.17%, 9.71%, 3.16%and 100%, 86.17%, 44.15%, 16.35%, 5.92%. Showing aged black garlic extract enhancing the sensitivity of radiotherapy to Huh7 cells.3. PI tested apoptosis as follow: 5.26%, 17.53%, 22.49%,29.32%. Such, certain concentration aged black garlic extract could promote Huh7 cells induce G2/M block and apoptosis. 4. Electron microscope showed increase concentration of aged black garlic extract can prominent formation of autophagic, lamellar structures, residual digested material.5. Up regulation expression of Beclinl and LC3, relationshiped with concentration of aged black garlic extract. Inhibitory rate of high, low concentration of aged black garlic extract plus radiation group and radiation group were 67.2%, 58.4%, 51.5%. Conclusion: 1. Aged black garlic could inhibit proliferation of liver cancer Huh7 cells, and sensitize the liver cancer Huh7 cells to ionizing irradiation. 2. Aged black garlic induce apoptosis and G2/M block. These maybe mechanism of ionizing irradiation. 3. The mechanism of sensitization may related to up regulating the expression of Beclinland LC3.