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A preliminary study on the radiation-resistance mechanism in ovarian cancer
Qi Liao1, Hong-mei Zhang, Hai-hua Li
1Department of Radiotherapy, Second Teaching Hospital of Jilin University, Changchun, China.
Aim:
The present study was designed to explore the radiation-resistance mechanism by interfering in checkpoints kinase 1 (CHK1) and DNA-activated protein kinase (DNA-PK) genes with short hairpin RNA (shRNA) transfection into Skov3 cells derived from ovarian cancer and HeLa cells derived from cervical cancer.
Materials And Methods:
The cultured Skov3 and HeLa cells were transfected with plasmid vectors containing CHK1 shRNA and DNA-PK shRNA, respectively, through Lipofectimine™ 2000 mediation, and cultured for 20 hours before exposure to 2 Gy X-radiation. The cells were harvested 4 and 28 after X-irradiation respectively then washed 3 times with PBS. These cells were stained with Annexin V/PI and applied by flow cytometer to analyze alteration of apoptosis with software CellQuest.
Results:
The apoptotic response in Skov3 cells to X-radiation was significantly lower than that in HeLa cells at 4 hour (t = 15.22, P < 0.001) and 28 hours (t = 15.78, P < 0.001) of post-irradiation. The shRNA might not affect the apoptosis of Skov3 and HeLa cells, while shRNA-transfection significantly enhanced the apoptotic response in Skov3 cells to X-radiation as compared with that in HeLa cells.
Conclusions:
The present work suggests that the CHK1 and DNA-PK genes are very likely to play a role in developing a radiation resistance in ovarian cancer.
Insights
Checkpoint kinase 1 (CHK1) and DNA-activated protein kinase (DNA-PK) gene interference using shRNA enhanced radiation-induced apoptosis in ovarian cancer cells, suggesting their role in radiation resistance.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Ovarian and cervical cancers exhibit varying responses to radiation therapy.
- Understanding the molecular mechanisms of radiation resistance is crucial for improving treatment efficacy.
Purpose of the Study:
- To investigate the role of Checkpoint Kinase 1 (CHK1) and DNA-Activated Protein Kinase (DNA-PK) in radiation resistance.
- To explore the effects of short hairpin RNA (shRNA) targeting CHK1 and DNA-PK on cancer cell apoptosis following radiation exposure.
Main Methods:
- Skov3 (ovarian) and HeLa (cervical) cancer cells were transfected with CHK1 and DNA-PK shRNA using Lipofectamine™ 2000.
- Cells were exposed to 2 Gy X-radiation and apoptosis was analyzed at 4 and 28 hours post-irradiation using Annexin V/PI staining and flow cytometry.
Main Results:
- Skov3 cells exhibited significantly lower apoptotic response to X-radiation compared to HeLa cells at both 4 and 28 hours post-irradiation.
- While shRNA alone did not significantly affect apoptosis, shRNA-transfection enhanced the radiation-induced apoptotic response in Skov3 cells relative to HeLa cells.
Conclusions:
- CHK1 and DNA-PK genes are implicated in the development of radiation resistance in ovarian cancer.
- Targeting CHK1 and DNA-PK may represent a potential strategy to overcome radiation resistance in ovarian cancer.
