DNA Damage of Glioblastoma Multiform Cells Induced by Beta Radiation of Iodine-131 in The Presence or Absence of Topotecan: A Picogreen and Colonogenic Assay
Objective: Glioblastoma multiforme (GBM), one of the most common and aggressive malignant brain tumors, is highly resistant to radiotherapy. Numerous approaches have been pursued to find new radiosensitizers. We used a picogreen and colonogenic assay to appraise the DNA damage and cell death in a...
| Main Authors: | , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Royan Institute (ACECR), Tehran
2015-04-01
|
| Series: | Cell Journal |
| Subjects: | |
| Online Access: | http://celljournal.org/library/upload/article/af_3355325282433372232258327453726446555387Cell-J-17-99.pdf |
| _version_ | 1819064191058706432 |
|---|---|
| author | Nazila Eyvazzadeh Ali Neshasteh-Riz Seyed Rabee Mahdavi |
| author_facet | Nazila Eyvazzadeh Ali Neshasteh-Riz Seyed Rabee Mahdavi |
| author_sort | Nazila Eyvazzadeh |
| collection | DOAJ |
| description | Objective: Glioblastoma multiforme (GBM), one of the most common and aggressive
malignant brain tumors, is highly resistant to radiotherapy. Numerous approaches have
been pursued to find new radiosensitizers. We used a picogreen and colonogenic assay
to appraise the DNA damage and cell death in a spheroid culture of GBM cells caused by
iodine-131 (I-131) beta radiation in the presence of topotecan (TPT).
Materials and Methods: U87MG cells were cultured as spheroids with approximate
diameters of 300 μm. Cells were treated with beta radiation of I-131 (at a dose of 2 Gy)
and/ or TPT (1 μg/ml for 2 hours). The numbers of cells that survived were compared with
untreated cells using a colonogenic assay. In addition, we evaluated possible DNA damages
by the picogreen method. The relation between DNA damage and cell death was
assessed in the experimental study of groups.
Results: The findings showed that survival fraction (SF) in the I-131+TPT group
(39%) was considerably less than the I-131 group (58.92%; p<0.05). The number of
single strand breaks (SSB) and double strand breaks (DSB), in the DNA of U87MG
cells treated with beta radiation of I-131 and TPT (I-131+TPT) significantly increased
compared to cells treated with only I-131 or TPT (p<0.05). The amount of SSB repair
was more than DSB repair (p<0.05). The relationship between cell death and
DNA damage was close (r≥0.6) and significant (p<0.05) in the irradiated and treated
groups. Also the maximum rate of DNA repair occurred 24 hours after the treatments.
A significant difference was not observed on other days of the restoration.
Conclusion: The findings in the present study indicated that TPT can sensitize
U87MG cells to radiation and increase DNA damages. Potentially, TPT can cause
an increase in damage from DSB and SSB by its inhibitory effects on topoisomerase
enzyme and the cell cycle. The increased complex damages following the use of a
genotoxic agent and beta I-131 radiation, causes a significant increase the cell death
because of the difficult repair process. By assessing the relationship between DNA
damage and cell death, the picogreen method can be useful in predicting colonogenic
assay. Consequently, it is suggested that co-treatment with I-131 beta radiation and
TPT can improve GBM treatment. |
| first_indexed | 2024-12-21T15:26:38Z |
| format | Article |
| id | doaj.art-c85063adb4fd410e888cd0b55e72145e |
| institution | Directory Open Access Journal |
| issn | 2228-5806 2228-5814 |
| language | English |
| last_indexed | 2024-12-21T15:26:38Z |
| publishDate | 2015-04-01 |
| publisher | Royan Institute (ACECR), Tehran |
| record_format | Article |
| series | Cell Journal |
| spelling | doaj.art-c85063adb4fd410e888cd0b55e72145e2022-12-21T18:58:53ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142015-04-0117199110DNA Damage of Glioblastoma Multiform Cells Induced by Beta Radiation of Iodine-131 in The Presence or Absence of Topotecan: A Picogreen and Colonogenic AssayNazila Eyvazzadeh0Ali Neshasteh-Riz1Seyed Rabee Mahdavi2Radiation Research Center, Faculty of Paramedicine, AJA University of Medical Sciences, Tehran, IranDepartment of Radiology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, IranDepartment of Medical Physics, Faculty of Medicine, Iran University of Medical Sciences, Tehran, IranObjective: Glioblastoma multiforme (GBM), one of the most common and aggressive malignant brain tumors, is highly resistant to radiotherapy. Numerous approaches have been pursued to find new radiosensitizers. We used a picogreen and colonogenic assay to appraise the DNA damage and cell death in a spheroid culture of GBM cells caused by iodine-131 (I-131) beta radiation in the presence of topotecan (TPT). Materials and Methods: U87MG cells were cultured as spheroids with approximate diameters of 300 μm. Cells were treated with beta radiation of I-131 (at a dose of 2 Gy) and/ or TPT (1 μg/ml for 2 hours). The numbers of cells that survived were compared with untreated cells using a colonogenic assay. In addition, we evaluated possible DNA damages by the picogreen method. The relation between DNA damage and cell death was assessed in the experimental study of groups. Results: The findings showed that survival fraction (SF) in the I-131+TPT group (39%) was considerably less than the I-131 group (58.92%; p<0.05). The number of single strand breaks (SSB) and double strand breaks (DSB), in the DNA of U87MG cells treated with beta radiation of I-131 and TPT (I-131+TPT) significantly increased compared to cells treated with only I-131 or TPT (p<0.05). The amount of SSB repair was more than DSB repair (p<0.05). The relationship between cell death and DNA damage was close (r≥0.6) and significant (p<0.05) in the irradiated and treated groups. Also the maximum rate of DNA repair occurred 24 hours after the treatments. A significant difference was not observed on other days of the restoration. Conclusion: The findings in the present study indicated that TPT can sensitize U87MG cells to radiation and increase DNA damages. Potentially, TPT can cause an increase in damage from DSB and SSB by its inhibitory effects on topoisomerase enzyme and the cell cycle. The increased complex damages following the use of a genotoxic agent and beta I-131 radiation, causes a significant increase the cell death because of the difficult repair process. By assessing the relationship between DNA damage and cell death, the picogreen method can be useful in predicting colonogenic assay. Consequently, it is suggested that co-treatment with I-131 beta radiation and TPT can improve GBM treatment.http://celljournal.org/library/upload/article/af_3355325282433372232258327453726446555387Cell-J-17-99.pdfDNA DamageTopotecanGlioblastomaCell Death |
| spellingShingle | Nazila Eyvazzadeh Ali Neshasteh-Riz Seyed Rabee Mahdavi DNA Damage of Glioblastoma Multiform Cells Induced by Beta Radiation of Iodine-131 in The Presence or Absence of Topotecan: A Picogreen and Colonogenic Assay Cell Journal DNA Damage Topotecan Glioblastoma Cell Death |
| title | DNA Damage of Glioblastoma Multiform Cells Induced by Beta Radiation of Iodine-131 in The Presence or Absence of Topotecan: A Picogreen and Colonogenic Assay |
| title_full | DNA Damage of Glioblastoma Multiform Cells Induced by Beta Radiation of Iodine-131 in The Presence or Absence of Topotecan: A Picogreen and Colonogenic Assay |
| title_fullStr | DNA Damage of Glioblastoma Multiform Cells Induced by Beta Radiation of Iodine-131 in The Presence or Absence of Topotecan: A Picogreen and Colonogenic Assay |
| title_full_unstemmed | DNA Damage of Glioblastoma Multiform Cells Induced by Beta Radiation of Iodine-131 in The Presence or Absence of Topotecan: A Picogreen and Colonogenic Assay |
| title_short | DNA Damage of Glioblastoma Multiform Cells Induced by Beta Radiation of Iodine-131 in The Presence or Absence of Topotecan: A Picogreen and Colonogenic Assay |
| title_sort | dna damage of glioblastoma multiform cells induced by beta radiation of iodine 131 in the presence or absence of topotecan a picogreen and colonogenic assay |
| topic | DNA Damage Topotecan Glioblastoma Cell Death |
| url | http://celljournal.org/library/upload/article/af_3355325282433372232258327453726446555387Cell-J-17-99.pdf |
| work_keys_str_mv | AT nazilaeyvazzadeh dnadamageofglioblastomamultiformcellsinducedbybetaradiationofiodine131inthepresenceorabsenceoftopotecanapicogreenandcolonogenicassay AT alineshastehriz dnadamageofglioblastomamultiformcellsinducedbybetaradiationofiodine131inthepresenceorabsenceoftopotecanapicogreenandcolonogenicassay AT seyedrabeemahdavi dnadamageofglioblastomamultiformcellsinducedbybetaradiationofiodine131inthepresenceorabsenceoftopotecanapicogreenandcolonogenicassay |