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Influence of P53 on the radiotherapy response of hepatocellular carcinoma.

Gomes AR, Abrantes AM, Brito AF, Laranjo M, Casalta-Lopes JE, Gonçalves AC, Sarmento-Ribeiro AB, Botelho MF, Tralhão JG - Clin Mol Hepatol (2015)

Bottom Line: Flow cytometry was carried out to examine the effects of iodine-131 on cell death, oxidative stress, reduced intracellular glutathione expression, the mitochondrial membrane potential, and the cell cycle.Irradiation induced a decrease in cell survival and led to a decrease in cell viability in all of the cell lines studied, accompanied by cell death via late apoptosis/necrosis and necrosis.Irradiation with 131-iodine induced mostly cell-cycle arrest in the G0/G1 phase.

View Article: PubMed Central - PubMed

Affiliation: Biophysics Unit, Faculty of Medicine of University of Coimbra, Coimbra, Portugal.

ABSTRACT

Background/aims: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, and it has a poor prognosis and few therapeutic options. Radiotherapy is one of the most effective forms of cancer treatment, and P53 protein is one of the key molecules determining how a cell responds to radiotherapy. The aim of this study was to determine the therapeutic efficacy of iodine-131 in three human HCC cell lines.

Methods: Western blotting was used to measure P53 expression. The effects of radiotherapy with iodine-131 were assessed by using the clonogenic assay to evaluate cell survival. Flow cytometry was carried out to examine the effects of iodine-131 on cell death, oxidative stress, reduced intracellular glutathione expression, the mitochondrial membrane potential, and the cell cycle.

Results: The P53 protein was not expressed in Hep3B2.1-7 cells, was expressed at normal levels in HepG2 cells, and was overexpressed in HuH7 cells. P53 expression in the HuH7 and HepG2 cell lines increased after internal and external irradiation with iodine-131. Irradiation induced a decrease in cell survival and led to a decrease in cell viability in all of the cell lines studied, accompanied by cell death via late apoptosis/necrosis and necrosis. Irradiation with 131-iodine induced mostly cell-cycle arrest in the G0/G1 phase.

Conclusions: These results suggest that P53 plays a key role in the radiotherapy response of HCC.

No MeSH data available.


Related in: MedlinePlus

Cell viability analysis using flow cytometry with AnV/IP double staining. The results are percentages of cells that are viable, in apoptosis, in late apoptosis/necrosis, and in necrosis at 24 hours after IR or ER with iodine-131. Data are mean and SE values (n=8). Asterisks indicate statistically significant differences compared to controls (P<0.05).
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Figure 5: Cell viability analysis using flow cytometry with AnV/IP double staining. The results are percentages of cells that are viable, in apoptosis, in late apoptosis/necrosis, and in necrosis at 24 hours after IR or ER with iodine-131. Data are mean and SE values (n=8). Asterisks indicate statistically significant differences compared to controls (P<0.05).

Mentions: Using internal irradiation, HepG2 cells showed 49% of viability (P=0.031), and a tendency to activate cell death by necrosis (P=0.03) at 20 Gy (Fig. 5). The HuH7 cells had 59% of viability after 1 Gy of irradiation, and 20% of cells died by apoptosis (P=0.02). The Hep3B2.1-7 cells had 49% of viability, and 24% died by apoptosis (P=0.003), after internal irradiation with 20 Gy. After 1 Gy external irradiation, the HepG2 cells had 40% of viability (P=0.013), and 28% of viability at 20 Gy (P=0.001). Regarding cell death, 32% of Hep3B2.1-7 cells died by necrosis with 1 Gy irradiation dose (P=0.017), and at 20 Gy, 25% died by late apoptosis/necrosis (P=0.016), and 35% by necrosis (P=0.031) (Fig. 5). The HuH7 cells showed 17% of death by late apoptosis/necrosis at 1 Gy and 20% at 20 Gy. The Hep3B2.1-7 cells had 49% of viability (P=0.036), 19% dying by late apoptosis/necrosis (P=0.024) and 20% by necrosis (P=0.046) at 20 Gy.


Influence of P53 on the radiotherapy response of hepatocellular carcinoma.

Gomes AR, Abrantes AM, Brito AF, Laranjo M, Casalta-Lopes JE, Gonçalves AC, Sarmento-Ribeiro AB, Botelho MF, Tralhão JG - Clin Mol Hepatol (2015)

Cell viability analysis using flow cytometry with AnV/IP double staining. The results are percentages of cells that are viable, in apoptosis, in late apoptosis/necrosis, and in necrosis at 24 hours after IR or ER with iodine-131. Data are mean and SE values (n=8). Asterisks indicate statistically significant differences compared to controls (P<0.05).
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4612287&req=5

Figure 5: Cell viability analysis using flow cytometry with AnV/IP double staining. The results are percentages of cells that are viable, in apoptosis, in late apoptosis/necrosis, and in necrosis at 24 hours after IR or ER with iodine-131. Data are mean and SE values (n=8). Asterisks indicate statistically significant differences compared to controls (P<0.05).
Mentions: Using internal irradiation, HepG2 cells showed 49% of viability (P=0.031), and a tendency to activate cell death by necrosis (P=0.03) at 20 Gy (Fig. 5). The HuH7 cells had 59% of viability after 1 Gy of irradiation, and 20% of cells died by apoptosis (P=0.02). The Hep3B2.1-7 cells had 49% of viability, and 24% died by apoptosis (P=0.003), after internal irradiation with 20 Gy. After 1 Gy external irradiation, the HepG2 cells had 40% of viability (P=0.013), and 28% of viability at 20 Gy (P=0.001). Regarding cell death, 32% of Hep3B2.1-7 cells died by necrosis with 1 Gy irradiation dose (P=0.017), and at 20 Gy, 25% died by late apoptosis/necrosis (P=0.016), and 35% by necrosis (P=0.031) (Fig. 5). The HuH7 cells showed 17% of death by late apoptosis/necrosis at 1 Gy and 20% at 20 Gy. The Hep3B2.1-7 cells had 49% of viability (P=0.036), 19% dying by late apoptosis/necrosis (P=0.024) and 20% by necrosis (P=0.046) at 20 Gy.

Bottom Line: Flow cytometry was carried out to examine the effects of iodine-131 on cell death, oxidative stress, reduced intracellular glutathione expression, the mitochondrial membrane potential, and the cell cycle.Irradiation induced a decrease in cell survival and led to a decrease in cell viability in all of the cell lines studied, accompanied by cell death via late apoptosis/necrosis and necrosis.Irradiation with 131-iodine induced mostly cell-cycle arrest in the G0/G1 phase.

View Article: PubMed Central - PubMed

Affiliation: Biophysics Unit, Faculty of Medicine of University of Coimbra, Coimbra, Portugal.

ABSTRACT

Background/aims: Hepatocellular carcinoma (HCC) is one of the most common cancers worldwide, and it has a poor prognosis and few therapeutic options. Radiotherapy is one of the most effective forms of cancer treatment, and P53 protein is one of the key molecules determining how a cell responds to radiotherapy. The aim of this study was to determine the therapeutic efficacy of iodine-131 in three human HCC cell lines.

Methods: Western blotting was used to measure P53 expression. The effects of radiotherapy with iodine-131 were assessed by using the clonogenic assay to evaluate cell survival. Flow cytometry was carried out to examine the effects of iodine-131 on cell death, oxidative stress, reduced intracellular glutathione expression, the mitochondrial membrane potential, and the cell cycle.

Results: The P53 protein was not expressed in Hep3B2.1-7 cells, was expressed at normal levels in HepG2 cells, and was overexpressed in HuH7 cells. P53 expression in the HuH7 and HepG2 cell lines increased after internal and external irradiation with iodine-131. Irradiation induced a decrease in cell survival and led to a decrease in cell viability in all of the cell lines studied, accompanied by cell death via late apoptosis/necrosis and necrosis. Irradiation with 131-iodine induced mostly cell-cycle arrest in the G0/G1 phase.

Conclusions: These results suggest that P53 plays a key role in the radiotherapy response of HCC.

No MeSH data available.


Related in: MedlinePlus