Anandamide induced autophagy and apoptosis in human hepatoma cells

[Speaker] Chun Chien Yang:1
[Co-author] Chia Ming Chang:1, Yun Hong Wong:1, Meng Hsuan Cheng:2, Yu Chiao Yang:1, Hsiao Mei Kuo:3, Hun Lung Yuan:4, Hui Fen Chiu:1
1:Pharmacology, Kaohsiung Medical University, Taiwan, 2:Department of Internal Medicine, Chung-Ho Memorial Hospital, Kaohsiung Medical University, Taiwan, 3:Department of Neuroscience, Marine Biotechnology and Resources, National Sun Yat-sen University, Taiwan, 4:Deparment of Chemistry, R. O. C. Military Academy, Kaohsiung, Taiwan

Backgrounds: Anandamide (AEA) is the one of endogenous cannabinoids (Endocannabinoids) from porcine brain. Recently, AEA have attracted more attention due to its effective anticancer properties in various cancer types. Autophagy is an evolutionarily conserved catabolic process involving lysosomal degradation of intracellular components and damage of organelles. However, the exact role of autophagy in cancer cells remains unclear. Our previous studies have investigate the anti-tumor activity of AEA was regulated by cell cycle G1 phase arrest in human hepatocellular carcinoma (HCC) cells. The current study, we further evaluated the role of autophagy in tumor suppressive effect of AEA in HCC.

Materials and Methods: The anti-proliferative effect of AEA was determined by methylene blue and XTT assay. Detection of autophagic vacuoles (acidic vesicular organelle) was conducted by immunofluorescent (IF) staining. Annexin V and propidium iodide (PI) double staining was used for apoptotic cell quantification. The autophagic and apoptotic related protein signal expression was detected by Western blotting analysis.

Results: Our study has observed that AEA processed cytotoxic activity and morphologic changes in a dose dependent manner in HCC cell line. AVO formation was observed in AEA-treated HCC cells. Flow cytometry data indicated that AEA treatment for 48 h significantly induced apoptotic cell death. Further Western blotting analysis revealed that AEA upregulated microtubule-associated protein light chain3 II (LC3-II), SQSTM1/p62, Bax and cleaved PARP protein expression.

Conclusion: In summary, Our results have indicated that AEA effectively inhibited HCC cell growth via autophagy and apoptosis induction.

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