Suppression of human 8-oxoguanine DNA glycosylase (OGG1) augments ultrasound-induced apoptosis in cervical cancer cells

Autor: Yongli Nie, Jiao Bai, Hongxia Jing, Xiongfei Cheng, Bo Yang, Tao Xu, Jun Zhang, Linjun Li, Guang-mei Zheng, Jiao Jiao, Jianyun Yu
Rok vydání: 2016
Předmět:
0301 basic medicine
Acoustics and Ultrasonics
Cell Survival
Ultrasonic Therapy
Blotting
Western
Down-Regulation
Uterine Cervical Neoplasms
Apoptosis
DNA Glycosylases
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
Bcl-2-associated X protein
Downregulation and upregulation
Annexin
Cell Line
Tumor
Humans
Viability assay
Propidium iodide
Annexin A5
RNA
Small Interfering
Cell Proliferation
bcl-2-Associated X Protein
Aniline Compounds
biology
Chemistry
Caspase 3
NF-kappa B
Fluoresceins
Molecular biology
Oxidative Stress
030104 developmental biology
Intrinsic apoptotic signaling pathway
Proto-Oncogene Proteins c-bcl-2
Xanthenes
030220 oncology & carcinogenesis
Immunology
biology.protein
Calcium
Female
Reactive Oxygen Species
Fluorescein-5-isothiocyanate
Signal Transduction
Zdroj: Ultrasonics. 72
ISSN: 1874-9968
Popis: Purpose Human 8-oxoguanine DNA glycosylase (OGG1) is a major base excision repair enzyme, and it was reported to suppress the activation of intrinsic apoptotic signaling pathway in response to oxidative stress. In this study, our aim was to investigate the effects of OGG1 downregulation on ultrasound-induced apoptosis in cervical cancer cells. Methods OGG1 expression was silenced by shRNA in the cervical cancer SW756 and CaSki cells. Cell viability was evaluated by MTT assay after OGG1 knockdown following ultrasound treatment. Ultrasound-induced apoptosis was measured by Annexin V-FITC/propidium iodide. Intracellular reactive oxygen species (ROS) production and Ca2+ concentration were detected using a fluorescent probe, 2′,7′-dichlorofluorescin diacetate (DCFH-DA) and a green fluorescent dye fluo-4AM, respectively. Western blotting was used to analyze the expression of Bcl-2, Bax, cleaved caspase-3, and nuclear factor-κB p65 (NF-κB p65). Results The results indicated that OGG1 knockdown did not suppress cell proliferation, but significantly augmented ultrasound-induced inhibitory effects on the cell viability, and increased ultrasound-induced early apoptosis and late apoptosis and necrosis in the SW756 and CaSki cells when exposure to ultrasound (1 MHz) at 1.5 W/cm2 for 30 and 60 s. OGG1 knockdown significantly increased intracellular ROS production and Ca2+ concentration after incubation of 6, 24, and 48 h post-ultrasound treatment. The downregulation of Bcl-2 protein and the upregulation of Bax, cleaved caspase-3, and NF-κB p65 protein levels were observed in the shRNA-OGG1 cells and mock-shRNA cells, but no significant change of these protein levels was found between of them. Conclusions These results indicate that downregulation of OGG1 expression can augment ultrasound-induced apoptosis in cervical cancer cells, which suggests that OGG1 suppression might provide a new insight for ultrasound-induced therapeutic effects on cervical cancer treatment.
Databáze: OpenAIRE
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