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Different effects of H2O2 treatment on cervical squamous carcinoma cells and adenocarcinoma cells.

Zhang P, Yin H, Wang S, Wei Y, Peng N, Bi W, Wang X - Arch Med Sci (2015)

Bottom Line: Reactive oxygen species levels and GSSG levels were significantly increased (p < 0.01), whereas GSH levels were significantly decreased (p < 0.05 or 0.01) in both cells after H2O2 treatment.Thus the ratio of GSH/GSSG was significantly decreased by H2O2 treatment in both cells (p < 0.01).In addition, H2O2 treatment significantly increased activities of SOD, CAT, and GPx in both cells (p < 0.05 or 0.01).

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Qilu Hospital, Shandong University, Jinan, Shandong, China.

ABSTRACT

Introduction: This study aims to compare the antioxidant abilities of cervical squamous carcinoma cells and cervical adenocarcinoma cells and to study the related mechanisms.

Material and methods: Cervical squamous carcinoma and adenocarcinoma cells were treated with H2O2. Cell proliferation was determined with the MTT assay. The reactive oxygen species (ROS) level was detected by the 2',7'-dichlorofluorescein-diacetate (DCFH-DA) method. The 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) method was performed to measure intracellular concentrations of reduced glutathione (GSH) and oxidized glutathione (GSSG). The nitrite formation method, the molybdate colorimetric method, and the DTNB colorimetric method were used to determine activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), respectively.

Results: Compared with untreated control cells, cell proliferation of cervical squamous carcinoma cells and cervical adenocarcinoma cells was significantly inhibited by H2O2 treatment (p < 0.05). Reactive oxygen species levels and GSSG levels were significantly increased (p < 0.01), whereas GSH levels were significantly decreased (p < 0.05 or 0.01) in both cells after H2O2 treatment. Thus the ratio of GSH/GSSG was significantly decreased by H2O2 treatment in both cells (p < 0.01). In addition, H2O2 treatment significantly increased activities of SOD, CAT, and GPx in both cells (p < 0.05 or 0.01). Furthermore, the above-mentioned changes induced by H2O2 treatment were more dramatic in cervical squamous carcinoma cells.

Conclusions: The antioxidant ability of cervical squamous carcinoma cells is lower than that of cervical adenocarcinoma cells, which may be related to the increased ROS levels in cervical squamous carcinoma cells induced by H2O2 treatments.

No MeSH data available.


Related in: MedlinePlus

Determination of the ROS, GSH, GSSG levels, as well as the GSH/GSSG ratios, in cells treated with or without H2O2. The cells were treated with or without H2O2 (250 μM) for 24 h. The ROS levels of those cells were measured with the DCFH-DA method. The DTNB method was applied for detection of intracellular GSH and GSSG content. A – ROS levels in cells. B – GSH levels in cells. C – GSSG levels in cells. D – GSH/GSSG ratio in cells. Compared with HeLa and GH329 cells treated with medium only, *p < 0.05, **p < 0.01. Compared with Siha and SW756 cells treated with medium only, #p < 0.05, ##p < 0.01
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Figure 0002: Determination of the ROS, GSH, GSSG levels, as well as the GSH/GSSG ratios, in cells treated with or without H2O2. The cells were treated with or without H2O2 (250 μM) for 24 h. The ROS levels of those cells were measured with the DCFH-DA method. The DTNB method was applied for detection of intracellular GSH and GSSG content. A – ROS levels in cells. B – GSH levels in cells. C – GSSG levels in cells. D – GSH/GSSG ratio in cells. Compared with HeLa and GH329 cells treated with medium only, *p < 0.05, **p < 0.01. Compared with Siha and SW756 cells treated with medium only, #p < 0.05, ##p < 0.01

Mentions: To investigate whether H2O2 affects the ROS levels of cells, the cells were treated with or without H2O2 (250 µM) for 24 h. The ROS levels were measured with the DCFH-DA method. As shown in Figure 2 A, ROS levels of all of these 4 types of cells were significantly increased by H2O2 treatments when compared with the untreated cells (p < 0.05 or 0.01). However, the ROS levels in Siha and SW756 cells were increased much more significantly, up to 1.9 and 2.3 times the levels in their control groups, respectively (p < 0.01). These results indicated that H2O2 treatments increase the ROS levels more effectively in the Siha and SW756 cells than in the HeLa and GH329 cells.


Different effects of H2O2 treatment on cervical squamous carcinoma cells and adenocarcinoma cells.

Zhang P, Yin H, Wang S, Wei Y, Peng N, Bi W, Wang X - Arch Med Sci (2015)

Determination of the ROS, GSH, GSSG levels, as well as the GSH/GSSG ratios, in cells treated with or without H2O2. The cells were treated with or without H2O2 (250 μM) for 24 h. The ROS levels of those cells were measured with the DCFH-DA method. The DTNB method was applied for detection of intracellular GSH and GSSG content. A – ROS levels in cells. B – GSH levels in cells. C – GSSG levels in cells. D – GSH/GSSG ratio in cells. Compared with HeLa and GH329 cells treated with medium only, *p < 0.05, **p < 0.01. Compared with Siha and SW756 cells treated with medium only, #p < 0.05, ##p < 0.01
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4697061&req=5

Figure 0002: Determination of the ROS, GSH, GSSG levels, as well as the GSH/GSSG ratios, in cells treated with or without H2O2. The cells were treated with or without H2O2 (250 μM) for 24 h. The ROS levels of those cells were measured with the DCFH-DA method. The DTNB method was applied for detection of intracellular GSH and GSSG content. A – ROS levels in cells. B – GSH levels in cells. C – GSSG levels in cells. D – GSH/GSSG ratio in cells. Compared with HeLa and GH329 cells treated with medium only, *p < 0.05, **p < 0.01. Compared with Siha and SW756 cells treated with medium only, #p < 0.05, ##p < 0.01
Mentions: To investigate whether H2O2 affects the ROS levels of cells, the cells were treated with or without H2O2 (250 µM) for 24 h. The ROS levels were measured with the DCFH-DA method. As shown in Figure 2 A, ROS levels of all of these 4 types of cells were significantly increased by H2O2 treatments when compared with the untreated cells (p < 0.05 or 0.01). However, the ROS levels in Siha and SW756 cells were increased much more significantly, up to 1.9 and 2.3 times the levels in their control groups, respectively (p < 0.01). These results indicated that H2O2 treatments increase the ROS levels more effectively in the Siha and SW756 cells than in the HeLa and GH329 cells.

Bottom Line: Reactive oxygen species levels and GSSG levels were significantly increased (p < 0.01), whereas GSH levels were significantly decreased (p < 0.05 or 0.01) in both cells after H2O2 treatment.Thus the ratio of GSH/GSSG was significantly decreased by H2O2 treatment in both cells (p < 0.01).In addition, H2O2 treatment significantly increased activities of SOD, CAT, and GPx in both cells (p < 0.05 or 0.01).

View Article: PubMed Central - PubMed

Affiliation: Department of Obstetrics and Gynecology, Qilu Hospital, Shandong University, Jinan, Shandong, China.

ABSTRACT

Introduction: This study aims to compare the antioxidant abilities of cervical squamous carcinoma cells and cervical adenocarcinoma cells and to study the related mechanisms.

Material and methods: Cervical squamous carcinoma and adenocarcinoma cells were treated with H2O2. Cell proliferation was determined with the MTT assay. The reactive oxygen species (ROS) level was detected by the 2',7'-dichlorofluorescein-diacetate (DCFH-DA) method. The 5,5'-dithiobis-2-nitrobenzoic acid (DTNB) method was performed to measure intracellular concentrations of reduced glutathione (GSH) and oxidized glutathione (GSSG). The nitrite formation method, the molybdate colorimetric method, and the DTNB colorimetric method were used to determine activities of superoxide dismutase (SOD), catalase (CAT), and glutathione peroxidase (GPx), respectively.

Results: Compared with untreated control cells, cell proliferation of cervical squamous carcinoma cells and cervical adenocarcinoma cells was significantly inhibited by H2O2 treatment (p < 0.05). Reactive oxygen species levels and GSSG levels were significantly increased (p < 0.01), whereas GSH levels were significantly decreased (p < 0.05 or 0.01) in both cells after H2O2 treatment. Thus the ratio of GSH/GSSG was significantly decreased by H2O2 treatment in both cells (p < 0.01). In addition, H2O2 treatment significantly increased activities of SOD, CAT, and GPx in both cells (p < 0.05 or 0.01). Furthermore, the above-mentioned changes induced by H2O2 treatment were more dramatic in cervical squamous carcinoma cells.

Conclusions: The antioxidant ability of cervical squamous carcinoma cells is lower than that of cervical adenocarcinoma cells, which may be related to the increased ROS levels in cervical squamous carcinoma cells induced by H2O2 treatments.

No MeSH data available.


Related in: MedlinePlus