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

摘要
图/表
参考文献
相关文章 (15)

全文: PDF (1626 KB) HTML (1 KB)
输出: BibTeX | EndNote (RIS)

摘要目的：检测亚毒性浓度5-羟基-4’-硝基-7-丙酰氧基金雀异黄素（HNPG）体外X线照射对骨肉瘤Saos-2细胞增殖抑制的增敏作用及其可能分子生物学机制。方法：体外培养骨肉瘤Saos-2细胞，用MTT法检测亚毒性浓度HNPG体外对X线抑制Saos-2细胞增殖的增敏作用；用FCM检测亚毒性浓度HNPG体外对X线诱导Saos-2细胞凋亡、活性氧表达、膜电位变化的增敏作用；用ELISA检测亚毒性浓度HNPG体外对X线调节Saos-2细胞SOD、CAT、GSH和MDA的增敏作用；用Western blot检测亚毒性浓度HNPG体外对X线调节Saos-2细胞bcl-2、bax、cyt-c和cleaved-caspase-3蛋白的增敏作用。结果：亚毒性浓度HNPG体外可增强X线抑制Saos-2细胞增殖作用，提高X线诱导Saos-2细胞凋亡率，同时增强X线对Saos-2细胞SOD、CAT、GSH下调，促进MDA、ROS上调，增强线粒体膜电下降，与NS组、HNPG组和X线组比较组间差异有统计学意义（P＜0.05）；伴随着bax、cyt-c和cleaved-caspase-3蛋白表达上调，bcl-2蛋白表达降低，与NS组、HNPG组和X线组比较组间差异有统计学意义（P＜0.05）。结论：亚毒性浓度HNPG在体外能增敏X线对骨肉瘤Saos-2细胞的增殖抑制作用，其机制可能与其促进X线调节细胞内SOD、CAT和GSH表达，上调活性氧含量，损伤线粒体内膜，诱导细胞线粒体途径凋亡相关。

	服务
	把本文推荐给朋友
	加入我的书架
	加入引用管理器
	E-mail Alert
	RSS
	作者相关文章
	赵学正
	高伟
	唐保永
	郑思化
	李利军

关键词 ：骨肉瘤, 5-羟基-4&rsquo, -硝基-7-丙酰氧基金雀异黄素, 活性氧, 细胞增殖, 细胞凋亡

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

通讯作者: 郑思化，副主任医师，Email：zhengsh2020@163.com。

作者简介: 赵学正（1983-），男，山东泰安人，主治医师，硕士。

链接本文:

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

[1] DOMOXOUDIS S, KOUKOURAKIS I M, GIAKZIDIS
A G, et al. Hypofractionated accelerated chemo-radiotherapy (Chemo-HypoAR) with Cisplatin and Liposomal doxorubicin for the treatment of patients with Uterine Sarcomas[J]. In Vivo, 2019, 33(5): 1621-1624.
[2] BAI J, LUO X. 5-Hydroxy-4'-Nitro-7-Propionyloxy-Genistein inhibited invasion and metastasis via inactivating Wnt/β-catenin signal pathway in human endometrial carcinoma Ji endometrial cells[J]. Med Sci Monit, 2018, 24: 3230-3243.
[3] BAI J, YANG B J, LUO X. Effects of 5-hydroxy-4'-nitro-7-propionyloxy-genistein on inhibiting proliferation and invasion via activating reactive oxygen species in human ovarian cancer A2780/DDP cells[J]. Oncol Lett, 2018, 15(4): 5227-5235.
[4] CLÉMENT-ZHAO A, TANGUY M L, COTTU P, et al. Toxicity of locoregional radiotherapy in combination with bevacizumab in patients with non-metastatic breast cancer (TOLERAB): Final long-term evaluation[J]. PLoS One, 2019, 14(8): e0221816.
[5] HART K, PATEL S, KOVOOR J. Spontaneous coronary artery dissection associated with anal cancer management with fluorouracil and radiotherapy[J]. Cureus, 2019, 11(6): e4979.
[6] OH E, ANDREWS K J, MCMULLEN L M, et al. Tolerance to stress conditions associated with food safety in Campylobacter jejuni strains isolated from retail raw chicken[J]. Sci Rep, 2019, 9(1): 11915-11928.
[7] MAJEWSKI M, OGNIK K, JUŚKIEWICZ J. The interaction between resveratrol and two forms of copper as carbonate and nanoparticles on antioxidant mechanisms and vascular function in Wistar rats[J]. Pharmacol Rep, 2019, 71(5): 862-869.
[8] CHAVES-FILHO A B, PINTO I F D, DANTAS L S, et al. Alterations in lipid metabolism of spinal cord linked to amyotrophic lateral sclerosis[J]. Sci Rep, 2019, 9(1): 11642.
[9] NOUAIRI I, JALALI K, ESSID S, et al. Alleviation of cadmium-induced genotoxicity and cytotoxicity by calcium chloride in faba bean (Vicia faba L. var. minor) roots[J]. Physiol Mol Biol Plants, 2019, 25(4): 921-931.
[10] LIM S W, SHIN Y J, LUO K, et al. Ginseng increases Klotho expression by FoxO3-mediated manganese superoxide dismutase in a mouse model of tacrolimus-induced renal injury[J]. Aging (Albany NY), 2019, 11(15): 5548-5569.
[11] LIU S, HE Y, SHI J, et al. Allicin attenuates myocardial ischemia reperfusion injury in rats by inhibition of inflammation and oxidative stress[J]. Transplant Proc, 2019, 51(6): 2060-2065.
[12] ARAVINDAN S, NATARAJAN M, HERMAN T S, et al. Molecular basis of 'hypoxic' breast cancer cell radio-sensitization: phytochemicals converge on radiation induced Rel signaling[J]. Radiat Oncol, 2013, 8(46): 1-12.
[13] LI J F, XIE L J, QIN L P, et al. Apoptosis gene reprograming of human peripheral blood mononuclear cells induced by radioiodine-131 (131I) irradiation[J]. Indian J Med Res, 2019, 149(5): 627-632.
[14] MA X, DENG J, CUI X, et al. Berberine exhibits antioxidative effects and reduces apoptosis of the vaginal epithelium in bacterial vaginosis[J]. Exp Ther Med, 2019, 18(3): 2122-2130.
[15] XIE Y, YAO FL, LI X. MicroRNA-361 regulates apoptosis of cardiomyocytes after ischemic-reperfusion injury[J]. Eur Rev Med Pharmacol Sci, 2019, 23(12): 5413-5421.
[16] MAURYA V, KUMAR P, CHAKRABORTI S, et al. Zolmitriptan attenuates hepatocellular carcinoma via activation of caspase mediated apoptosis[J]. Chem Biol Interact, 2019, 308: 120-129.
[17] SAHIN I, BILIR B, ALI S, et al. Soy Isoflavones in integrative oncology: increased efficacy and decreased toxicity of cancer therapy[J]. Integr Cancer Ther, 2019, 1(18): 1-11.