亚毒性浓度HNPG体外对X线照射人骨肉瘤Saos-2细胞增殖抑制的增敏作用及机制

doi:10.3969/j.issn.2095-9400.2020.02.005

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Abstract Objective: To investigate the sensitization effect of sub-toxic concentration of 5-hydroxy- 4’-nitro-7-propionyloxy-genistein (HNPG) on radiotherapy for human osteosarcoma Saos-2 cells and its possible molecular mechanism. Methods: Saos-2 cells were cultured in vitro, and the inhibiting effect of sub-toxic concentration of HNPG and the sub-toxic dose of X-ray on Saos-2 cells were selected using MTT assay. The sensitization effect of HNPG on X-ray induced apoptosis were observed using flow cytometry with annexin V-FITC (AV) and propidium iodide (PI) staining. The sensitization effect of HNPG on X-ray regulating reactive oxygen species (ROS) were measured in flow cytometry with 2’, 7’‑dichlorodihydrofluorescein diacetate (DCFHDA) staining. The sensitization effect of HNPG on X ray regulating Mitochondrial Membrane Potential (MMP) were measured in flow cytometry with lipophilic cationic dye 2 (6 Amin 3 imino 3H xanthen 9 yl) benzoic acid methyl ester (Rh123) staining. The bioactivity of superoxide dismutase (SOD) and catalase (CAT) and the content of glutathione (GSH) and malondialdehyde (MDA) were detected by Enzyme-linked immunosorbent assay (Elisa). The related apoptotic proteins, such as bcl-2, bax, cyt-c and cleaved-caspase-3 were assessed using Western blot. Results: The sub-toxic concentration of HNPG could dramatically enhance the anti-tumor activity of the sub-toxic dose of X-ray against Saos-2 cells in vitro, inhibited the cells proliferation, induced the cells apoptosis, accompanied by the bioactivity of SOD, declined CAT, reduced content of GSH, and enhanced content of MDA. Simultaneously the content of ROS was increased and the level of MMP was cut down, accompanied by the up-regulated proteins of bax, cyt-c and cleaved-caspase-3 and down-regulated bcl-2 protein. Conclusion: The sub-toxic concentration of HNPG could sensitize the inhibiting proliferative effect of the sub-toxic dose of X ray in Saos-2 cells in vitro, which might be related to the decreased bioactivity of SOD and CAT, reduced content of GSH, accumulated ROS in cells, damaged integrity of mitochondrial membrane and induced cells apoptosis.

Key words： osteosarcoma 5-hydroxy-4’-nitro-7-propionyloxy-genistein reactive oxygen species cell proliferation cell apoptosis

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	ZHAO Xuezheng
	Gao Wei
	TANG Baoyong
	ZHENG Sihua
	LI Lijun

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https://xb.wmu.edu.cn/EN/10.3969/j.issn.2095-9400.2020.02.005 OR https://xb.wmu.edu.cn/EN/Y2020/V50/I2/108

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