Role of autophagy induced by anthocyanins in protection of DNA damages caused by UVB irradiation in HepG2 cells

Document Type : Novel Research Articles

Authors

1 Zoology dept., faculty of science, Assiut univ., Assiut, Egypt

2 Zoology Department, Faculty of Science, Assiut University, Egypt

3 Zoology dept, faculty of science, Assiut univ., Assiut, Egypt

4 Zoology dept, faculty of science, Assiut Univ., Assiut, Egypt

Abstract

DNA-protein cross-links (DPCs) and Thymidine dimer (Td) are special types of DNA damages that formed when DNA exposed to many inducers as UV irradiations. Anthocyanins are members of the flavonoid group of phytochemicals found in many plants including Hibiscus sabdariffa. It was reported that anthocyanins protect DNA from UV-induced damages. Here we investigate the role of autophagy induced by anthocyanins in protection of UVB-induced DPCs and Td in HepG2 cells. Cells were treated either with anthocyanins (anth), Rapamycin (Rap) or combination of anth and Bafilomycin A1 (Baf A1) or anth and 3-methyladenine (3Ma) for 24 h and then subjected to 0.36 J/cm2 UVB irradiation. Then DNA extraction for DPCs and Td investigation was carried out or cells were treated with Acridine orange (AO) for examination of autophagy. The results reveled that anth induces autophagy formation as detected by AO pH dependent green to red (G/R) color shifting. To investigate the role of anth-induced autophagy on DPCs and Td formation after UVB exposure, electrophoresis of protein covalently linked to DNA or Td dot plot analysis of DNA were carried out. It was found that anth treated cells have reduced DPCs and Td compared with the anth untreated cells. Treatment of cells with autophagy inhibitors Baf A1 or 3Ma together with anth significantly increased the DPCs and Td formation compared with anth treated cells. Also, Rap treatment alone protects cells from DPCs and Td formation which confirm the role of autophagy in protection of cells from UVB-induced DPCs and Td formation.

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