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Published on: August 21, 2013
Differential PARP cleavage: an indication for existence of multiple forms of cell death in human gliomas
Vasantha Kumar Bhaskara1, Sundaram Challa, Manas Panigrahi
1Department of Biotechnology, School of Life Sciences, University of Hyderabad, Hyderabad, AP, India.
Background:
Gliomas represent a diverse range of clinical presentation, histological differentiation, and response to therapy. Altered cell proliferation and cell death signals in gliomas are of great interest to elucidate the key molecules involved and to find effective treatment modalities. By considering the role of different proteases in correlation with differential poly (ADP-ribose) polymerase (PARP) fragmentation we have studied the pattern of cell death in human glioma tissues.
Materials And Methods:
In our study, five different human glioma biopsies were collected and analyzed for the PARP cleavage pattern by using western immunoblotting. Samples were also analyzed for pro-caspase 3, calpain I (micro) and II (m), granzyme-B and apoptosis-inducing factor (AIF). Parallel sections of histologically confirmed astrocytoma and glioblastoma multiforme (GBM) were used for immunohistochemical analysis of cleaved caspase-3, granzyme B, AIF and cyclo-oxygenase -2 (cox-2).
Results:
We found PARP fragmentation, along with usual approximately 89 kDa and approximately 24 kDa fragments, into other fragments of different molecular weights. Caspase mediated cell death may lead to appearance of larger approximately 89 kDa fragment and smaller approximately 24 kDa fragment indicating existence of apoptosis in the tumors. However, other fragments corresponding to approximately 64 kDa, approximately 54 kDa, and approximately 40 kDa were observed concomitantly in all glial tumor tissues.
Conclusions:
These results may indicate, not only apoptosis and necrosis, but there occurs the co-existence of intermediate cell death pathways in human glial tumors.
Insights
Human glioma tissues exhibit poly (ADP-ribose) polymerase (PARP) fragmentation, suggesting that cell death pathways beyond apoptosis and necrosis coexist in these tumors. This finding offers new insights into glioma progression and potential therapeutic targets.
Area of Science:
- Neuro-oncology
- Molecular Biology
- Cell Death Research
Background:
- Gliomas exhibit diverse clinical and histological features, impacting treatment response.
- Understanding altered cell proliferation and death signals is crucial for identifying therapeutic targets.
- Investigating poly (ADP-ribose) polymerase (PARP) fragmentation patterns can elucidate cell death mechanisms in gliomas.
Purpose of the Study:
- To analyze the pattern of cell death in human glioma tissues.
- To correlate protease activity with differential poly (ADP-ribose) polymerase (PARP) fragmentation.
- To investigate the co-existence of various cell death pathways in human glial tumors.
Main Methods:
- Western immunoblotting was used to analyze PARP cleavage patterns in five human glioma biopsies.
- Pro-caspase 3, calpains, granzyme-B, and apoptosis-inducing factor (AIF) were assessed.
- Immunohistochemical analysis of cleaved caspase-3, granzyme B, AIF, and cyclo-oxygenase-2 (COX-2) was performed on astrocytoma and glioblastoma multiforme (GBM) sections.
Main Results:
- PARP fragmentation revealed fragments beyond the typical 89 kDa and 24 kDa, including 64 kDa, 54 kDa, and 40 kDa fragments.
- The presence of 89 kDa and 24 kDa fragments suggests caspase-mediated apoptosis.
- Concomitant observation of additional fragments indicates complex cell death processes.
Conclusions:
- Human glial tumors display poly (ADP-ribose) polymerase (PARP) fragmentation patterns indicative of multiple cell death mechanisms.
- Results suggest the co-existence of apoptosis, necrosis, and intermediate cell death pathways.
- This finding has implications for understanding glioma biology and developing novel therapeutic strategies.
