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Peripheral Blood Lymphocytes as In Vitro Model to Evaluate Genomic Instability Caused by Low Dose Radiation

  • Tewari, Shikha (Pathology, Dr. Ram Manohar Lohia Institute of Medical Sciences) ;
  • Khan, Kainat (Pathology, Dr. Ram Manohar Lohia Institute of Medical Sciences) ;
  • Husain, Nuzhat (Pathology, Dr. Ram Manohar Lohia Institute of Medical Sciences) ;
  • Rastogi, Madhup (Radiology, Dr. Ram Manohar Lohia Institute of Medical Sciences) ;
  • Mishra, Surendra P (Radiology, Dr. Ram Manohar Lohia Institute of Medical Sciences) ;
  • Srivastav, Anoop K (Radiology, Dr. Ram Manohar Lohia Institute of Medical Sciences)
  • Published : 2016.06.01

Abstract

Diagnostic and therapeutic radiation fields are planned so as to reduce side-effects while maximising the dose to site but effects on healthy tissues are inevitable. Radiation causes strand breaks in DNA of exposed cells which can lead to chromosomal aberrations and cause malfunction and cell death. Several researchers have highlighted the damaging effects of high dose radiation but still there is a lacuna in identifying damage due to low dose radiation used for diagnostic purposes. Blood is an easy resource to study genotoxicity and to estimate the effects of radiation. The micronucleus assay and chromosomal aberration can indicate genetic damage and our present aim was to establish these with lymphocytes in an in vitro model to predict the immediate effects low dose radiation. Blood was collected from healthy individuals and divided into 6 groups with increasing radiation dose i.e., 0Gy, 0.10Gy, 0.25Gy, 0.50Gy, 1Gy and 2Gy. The samples were irradiated in duplicates using a LINAC in the radiation oncology department. Standard protocols were applied for chromosomal aberration and micronucleus assays. Metaphases were stained in Giemsa and 200 were scored per sample for the detection of dicentric or acentric forms. For micronuclei detection, 200 metaphases. Giemsa stained binucleate cells per sample were analysed for any abnormality. The micronuclei (MN) frequency was increased in cells exposed to the entire range of doses (0.1-2Gy) delivered. Controls showed minimal MN formation ($2.0%{\pm}0.05$) with triple MN ($5.6%{\pm}2.0$) frequency at the lowest dose. MN formation increased exponentially with the radiation dose thereafter with a maximum at 2Gy. Significantly elevated numbers of dicentric chromosomes were also observed, even at doses of 0.1-0.5Gy, compared to controls, and acentric chromosomes were apparent at 2Gy. In conclusion we can state that lymphocytes can be effectively used to study direct effect of low dose radiation.

Keywords

DNA damage;low dose radiation;CBMN assay;chromosomal aberrations;in vitro

Acknowledgement

Supported by : Science and Engineering Research Board (SERB)

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