Page 328 - 2014 Printable Abstract Book
P. 328
(PS6-06) Studies on effects of Ionizing Radiation on expression of Nrf2 and its downstream targets with
special reference to both molecular and enzymatic antioxidants in THP-1 cells. Shikha Mohan, Master of
2
2
1
2
Science-Biotechnology ; Damodar Gupta ; Sweta Sanguri ; and Mitu Lal , Institute of Nuclear Medicine
1
and Allied Sciences, Ghaziabad, Uttar Pradesh, India and Institute of Nuclear Medicine and Allied
Sciences, Delhi, India
2
Introduction: Nrf2 plays a central role in the response to various types of stress by activating a
myriad of cellular antioxidants, detoxification enzymes, drug efflux pumps, and other cyto-protective
proteins. It has been recognized as an important regulator of inflammation. Recently, Nrf2 has been
reported to induce DNA damage repair in cells under oxidative stress through direct binding to the
promoter regions of components of repair-some. Methods: THP-1 cells were used as a model to study
effect of radiation on monocyte cells. The cell viability was analysed by PI exclusion as a measure of
radiation sensitivity. Irradiated sample was processed at different time interval to study time and dose
kinetics. Since radiation exposure increases the level of reactive species, measurement of super- oxide
and nitric oxide in specific and ROS in general was done by DHE uptake, diazotization of Griess Reagent
and oxidation of CMH2DCFDA respectively. The levels of acid soluble thiols and activities of SOD, GR, GST
was also determined following exposure to ionising radiation. Results & Discussion: Following exposure
of THP-1 cells with γ radiation the viability were found to be reduced in dose and time dependent manner.
Moreover, dose dependent increase in generation of ROS were observed till 48hr which was decreased
there after (72hrs).Expression of HO-1 was found to be increased following low dose radiation exposure
to the cells. The translocation of Nrf2 was maximally observed was observed as early as 6 hours.
Moreover, levels of antioxidant like GR, GST, and SOD1 were also found to be significantly higher with
respect to control. The present study will be utilized in assessment of radio-modulation of novel Nrf2
activators in THP-1 cells.
(PS6-07) Use of next-generation sequencing to identify microRNA from rat blood and plasma as
1
1
1
biomarkers for radiation-induced pneumonitis. Feng Gao ; Jayashree Narayanan ; Brian Fish ; Pengyuan
1
1
1
2;1
1;2
Liu ; Yong Liu ; Mingyu Liang ; Elizabeth Jacobs ; and Meetha Medhora , Medical College of Wisconsin,
1
Milwaukee, WI and Dept of Veterans Affairs, Zablocki VAMC, Milwaukee, WI
2
Introduction: MicroRNA are altered during development, cancer, pathological conditions and
radiation. We have developed a model of whole thoracic irradiation in rats to induce pneumonitis.
Symptoms of pneumonitis first appear about 6 weeks after irradiation. Our goal is to identify circulating
microRNA in rat as biomarkers to predict radiation pneumonitis at or before 5 weeks when we are still
able to mitigate radiation pneumonitis using the drug enalapril. Methods: Wistar female rats
(WAG/RijCmcr) at 9-10 weeks of age were exposed to a single dose of 15 Gy of X-irradiation to the thorax
only, at the dose rate of 1.43 Gy/min. Irradiated animals were kept in same living condition as un-
irradiated controls. Irradiated rats were euthanized at 1 week, 2 weeks, 3 weeks and 4 weeks (n=9/group)
after radiation, with matched controls being euthanized at 1 week and 4 weeks. Lung tissue and blood
were harvested. RNA was extracted from fresh lung, blood and plasma by Trizol. Results: MicroRNA
libraries (48 total) were generated using Illumina TruSeq small RNA kit. Illumina analysis tool CASAVA was
used to identify microRNA from the data derived by next-generation sequencing. We have identified 574
mature microRNA and 373 new microRNA by the next-generation sequencing. Candidates from blood and
plasma were ranked by testing differential expression using DESeq. MicroRNA with >2 fold differences
326 | P a g e
special reference to both molecular and enzymatic antioxidants in THP-1 cells. Shikha Mohan, Master of
2
2
1
2
Science-Biotechnology ; Damodar Gupta ; Sweta Sanguri ; and Mitu Lal , Institute of Nuclear Medicine
1
and Allied Sciences, Ghaziabad, Uttar Pradesh, India and Institute of Nuclear Medicine and Allied
Sciences, Delhi, India
2
Introduction: Nrf2 plays a central role in the response to various types of stress by activating a
myriad of cellular antioxidants, detoxification enzymes, drug efflux pumps, and other cyto-protective
proteins. It has been recognized as an important regulator of inflammation. Recently, Nrf2 has been
reported to induce DNA damage repair in cells under oxidative stress through direct binding to the
promoter regions of components of repair-some. Methods: THP-1 cells were used as a model to study
effect of radiation on monocyte cells. The cell viability was analysed by PI exclusion as a measure of
radiation sensitivity. Irradiated sample was processed at different time interval to study time and dose
kinetics. Since radiation exposure increases the level of reactive species, measurement of super- oxide
and nitric oxide in specific and ROS in general was done by DHE uptake, diazotization of Griess Reagent
and oxidation of CMH2DCFDA respectively. The levels of acid soluble thiols and activities of SOD, GR, GST
was also determined following exposure to ionising radiation. Results & Discussion: Following exposure
of THP-1 cells with γ radiation the viability were found to be reduced in dose and time dependent manner.
Moreover, dose dependent increase in generation of ROS were observed till 48hr which was decreased
there after (72hrs).Expression of HO-1 was found to be increased following low dose radiation exposure
to the cells. The translocation of Nrf2 was maximally observed was observed as early as 6 hours.
Moreover, levels of antioxidant like GR, GST, and SOD1 were also found to be significantly higher with
respect to control. The present study will be utilized in assessment of radio-modulation of novel Nrf2
activators in THP-1 cells.
(PS6-07) Use of next-generation sequencing to identify microRNA from rat blood and plasma as
1
1
1
biomarkers for radiation-induced pneumonitis. Feng Gao ; Jayashree Narayanan ; Brian Fish ; Pengyuan
1
1
1
2;1
1;2
Liu ; Yong Liu ; Mingyu Liang ; Elizabeth Jacobs ; and Meetha Medhora , Medical College of Wisconsin,
1
Milwaukee, WI and Dept of Veterans Affairs, Zablocki VAMC, Milwaukee, WI
2
Introduction: MicroRNA are altered during development, cancer, pathological conditions and
radiation. We have developed a model of whole thoracic irradiation in rats to induce pneumonitis.
Symptoms of pneumonitis first appear about 6 weeks after irradiation. Our goal is to identify circulating
microRNA in rat as biomarkers to predict radiation pneumonitis at or before 5 weeks when we are still
able to mitigate radiation pneumonitis using the drug enalapril. Methods: Wistar female rats
(WAG/RijCmcr) at 9-10 weeks of age were exposed to a single dose of 15 Gy of X-irradiation to the thorax
only, at the dose rate of 1.43 Gy/min. Irradiated animals were kept in same living condition as un-
irradiated controls. Irradiated rats were euthanized at 1 week, 2 weeks, 3 weeks and 4 weeks (n=9/group)
after radiation, with matched controls being euthanized at 1 week and 4 weeks. Lung tissue and blood
were harvested. RNA was extracted from fresh lung, blood and plasma by Trizol. Results: MicroRNA
libraries (48 total) were generated using Illumina TruSeq small RNA kit. Illumina analysis tool CASAVA was
used to identify microRNA from the data derived by next-generation sequencing. We have identified 574
mature microRNA and 373 new microRNA by the next-generation sequencing. Candidates from blood and
plasma were ranked by testing differential expression using DESeq. MicroRNA with >2 fold differences
326 | P a g e
