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Low Doses of Cisplatin Induce Gene Alterations, Cell Cycle Arrest, and Apoptosis in Human Promyelocytic Leukemia Cells

View Article: PubMed Central - PubMed

ABSTRACT

Cisplatin is a known antitumor drug, but its mechanisms of action are not fully elucidated. In this research, we studied the anticancer potential of cisplatin at doses of 1, 2, or 3 µM using HL-60 cells as a test model. We investigated cisplatin effects at the molecular level using RNA sequencing, cell cycle analysis, and apoptotic assay after 24, 48, 72, and 96 hours of treatment. The results show that many genes responsible for molecular and cellular functions were significantly altered. Cisplatin treatment also caused the cells to be arrested at the DNA synthesis phase, and as the time increases, the cells gradually accumulated at the sub-G1 phase. Also, as the dose increases, a significant number of cells entered into the apoptotic and necrotic stages. Altogether, the data show that low doses of cisplatin significantly impact the viability of HL-60 cells, through modulation of gene expression, cell cycle, and apoptosis.

No MeSH data available.


Percentages of apoptosis and necrosis in HL-60 cells exposed to cisplatin. HL-60 cells were kept untreated (control) or treated with 1, 2, or 3 µM of cisplatin for 24, 48, 72, or 96 hours, incubated with Annexin V and PI, and analyzed using Cellometer Vision as described in the “Methods” section. The results are presented in bar graphs. Each graph represents the data for apoptosis/necrosis for 24-hour (A), 48-hour (B), 72-hour (C), and 96-hour (D) treatments. P < 0.05 was considered as statistically significant.
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f3-bmi-11-2016-113: Percentages of apoptosis and necrosis in HL-60 cells exposed to cisplatin. HL-60 cells were kept untreated (control) or treated with 1, 2, or 3 µM of cisplatin for 24, 48, 72, or 96 hours, incubated with Annexin V and PI, and analyzed using Cellometer Vision as described in the “Methods” section. The results are presented in bar graphs. Each graph represents the data for apoptosis/necrosis for 24-hour (A), 48-hour (B), 72-hour (C), and 96-hour (D) treatments. P < 0.05 was considered as statistically significant.

Mentions: To further analyze the cells for apoptotic/necrosis properties, the cisplatin-treated cells were stained with PI and Annexin V. Annexin V specifically binds phosphatidylserine in cell membrane in which the cells entered into apoptotic phase, whereas PI stains DNA in necrotic cells where the cell membrane disintegrates. The results show that cisplatin induced apoptosis/necrosis in HL-60 cells in a time- and dose-dependent manner. In 24-hour treatment, at doses of 1, 2, or 3 µM, apoptosis was induced by approximately 7%, 10%, or 12% compared to the control cells, respectively (Fig. 2). As the incubation period increases (48, 72, or 96 hours), apoptotic/necrotic cells increased proportionally with the doses. At the end of 96-hour treatment, apoptotic/necrotic cells were significantly increased in all the tested doses compared to the controls (Fig. 3). These results support the evidence that the accumulation of cisplatin-treated cells in sub-G1 phase of cell cycle is due to the cells entering into apoptotic or necrotic phases.


Low Doses of Cisplatin Induce Gene Alterations, Cell Cycle Arrest, and Apoptosis in Human Promyelocytic Leukemia Cells
Percentages of apoptosis and necrosis in HL-60 cells exposed to cisplatin. HL-60 cells were kept untreated (control) or treated with 1, 2, or 3 µM of cisplatin for 24, 48, 72, or 96 hours, incubated with Annexin V and PI, and analyzed using Cellometer Vision as described in the “Methods” section. The results are presented in bar graphs. Each graph represents the data for apoptosis/necrosis for 24-hour (A), 48-hour (B), 72-hour (C), and 96-hour (D) treatments. P < 0.05 was considered as statistically significant.
© Copyright Policy - open-access
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC4998075&req=5

f3-bmi-11-2016-113: Percentages of apoptosis and necrosis in HL-60 cells exposed to cisplatin. HL-60 cells were kept untreated (control) or treated with 1, 2, or 3 µM of cisplatin for 24, 48, 72, or 96 hours, incubated with Annexin V and PI, and analyzed using Cellometer Vision as described in the “Methods” section. The results are presented in bar graphs. Each graph represents the data for apoptosis/necrosis for 24-hour (A), 48-hour (B), 72-hour (C), and 96-hour (D) treatments. P < 0.05 was considered as statistically significant.
Mentions: To further analyze the cells for apoptotic/necrosis properties, the cisplatin-treated cells were stained with PI and Annexin V. Annexin V specifically binds phosphatidylserine in cell membrane in which the cells entered into apoptotic phase, whereas PI stains DNA in necrotic cells where the cell membrane disintegrates. The results show that cisplatin induced apoptosis/necrosis in HL-60 cells in a time- and dose-dependent manner. In 24-hour treatment, at doses of 1, 2, or 3 µM, apoptosis was induced by approximately 7%, 10%, or 12% compared to the control cells, respectively (Fig. 2). As the incubation period increases (48, 72, or 96 hours), apoptotic/necrotic cells increased proportionally with the doses. At the end of 96-hour treatment, apoptotic/necrotic cells were significantly increased in all the tested doses compared to the controls (Fig. 3). These results support the evidence that the accumulation of cisplatin-treated cells in sub-G1 phase of cell cycle is due to the cells entering into apoptotic or necrotic phases.

View Article: PubMed Central - PubMed

ABSTRACT

Cisplatin is a known antitumor drug, but its mechanisms of action are not fully elucidated. In this research, we studied the anticancer potential of cisplatin at doses of 1, 2, or 3 &micro;M using HL-60 cells as a test model. We investigated cisplatin effects at the molecular level using RNA sequencing, cell cycle analysis, and apoptotic assay after 24, 48, 72, and 96 hours of treatment. The results show that many genes responsible for molecular and cellular functions were significantly altered. Cisplatin treatment also caused the cells to be arrested at the DNA synthesis phase, and as the time increases, the cells gradually accumulated at the sub-G1 phase. Also, as the dose increases, a significant number of cells entered into the apoptotic and necrotic stages. Altogether, the data show that low doses of cisplatin significantly impact the viability of HL-60 cells, through modulation of gene expression, cell cycle, and apoptosis.

No MeSH data available.