LIFE09200604007 Tabish - Homi Bhabha National Institute

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Results Fig. 31: Percent cell death after BPDE exposure in MPN (n=20) and control cell lines (n=10). Percent cell death after 5μM radiation exposure (a) percent cell death after 8μM radiation exposure (b). Mean difference between percent relative cell death between MPN and control cell lines was statistically significant (p=0.0009 at 5μM and p=0.0008 at 8μM). The values shown are the mean values from three separate experiments. Table 12: Average cell death after γ-radiation and BPDE exposure in MPN and control cell lines Genotoxic exposure Dose/concent ration MPN cell line (n=20) Control cell line (n=10) Radiation exposure 5 Gy 10 Gy 9.84% (Range 8.3% - 12.7%) 10.84% (Range 8.3% - 12.7%) 19.22% (Range 9.05% - 42.82%) 22.09% (Range 12.04% - 42.4%) BPDE treatment 5 µM 13.57% (Range 8.02% - 21.77%) 8 µM 14.02% (Range 8.46% - 24.69%) 23.25% (Range 10.49% - 41.79%) 24.98% (Range 10.23% - 51.76%) 114
Results Fig. 32: Comparison of percent cell death between MPN and control groups. Percent cell death after genotoxic exposure is more pronounced in control group as compared to MPN group after γ-radiation. The difference was statistically significant between the two groups with *p=0.0003 at 5 Gy and **p=0.0002 at 10 Gy (a) and BPDE exposure with *p=0.0009 at 5μM and **p=0.0009 at 8μM BPDE (b). Vertical line on each bar shows standard deviation. 4.2.5 Global gene expression profiling after γ-radiation exposure It is well established that the way in which an individual deals with genotoxic exposure is related to the probability to develop cancer. Sensitivity to genotoxic agents in terms of phenotypic responses can reflect and individual‟s susceptibility to neoplastic evolution. At present, apart from performing standard tests to assess genotoxic effect the focus has started to shift to search for pathways involved in differential genotoxic sensitivity and phenotypic responses. These differences in cellular responses can be traced back to differential gene expression hence whole genome expression profiling was done in order to find any association between varied phenotypic response after genotoxic exposure and gene expression in patient and control cell lines. 4.2.5.1 Gene expression data analysis To assess the effect of γ-radiation exposure between MPN and control cell lines whole genome expression profiling was done using randomly selected 5 MPN and 5 control cell lines, at 5 Gy radiation exposure and 24 h time point. After processing the slides as described in Materials and Methods (Toronto microarray centre slides 19K and 8K were used) they were analyzed on AXON Scanner 4200A and raw data was obtained as ratio of two fluorescent dyes for each spot, corresponding to different genes along with other parameters. This data was subjected to analysis using GeneSpring GX 10.0 software. The raw data was first normalised by Lowess 115
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Genotype-Molecular Phenotype Correl
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CONTENTS Page No Synopsis 1 List of
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Synopsis
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Synopsis Genotype-Molecular Phenoty
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Synopsis Materials and Methods: 1.
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Synopsis loading control. PCR produ
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Synopsis 7.1 Percent cell death aft
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Synopsis slides were then dried and
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Synopsis range 41.77% - 90.95% at 5
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Synopsis 5. Genotyping of genes: Ge
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Synopsis G2/M arrest and delaying e
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Synopsis may have an important bear
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Synopsis References: 1. Bobba, R.;
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Abbreviations MOPS : 3-(N-morpholin
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List of Figures Fig. 28: Percent re
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Chapter 1 Introduction
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Introduction In previous studies fr
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Introduction important carcinogenes
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Review of Literature The yet undeci
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Review of Literature Fig. 2: Incide
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Review of Literature sustained angi
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Review of Literature 2.6 Genotype p
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Review of Literature 2.7.2 Biologic
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Review of Literature Table 1: Steps
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Review of Literature future analysi
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Review of Literature Inefficient ce
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Review of Literature XRCC1 (X-ray r
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Review of Literature 2.9.2 Gene inv
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Review of Literature MPO gene polym
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Review of Literature important role
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Materials and Methods Source of che
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Materials and Methods Method: EBV-t
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Materials and Methods Immunophenoty
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Materials and Methods with 500 μl
Page 73 and 74: Materials and Methods cDNA it can b
Page 75 and 76: Materials and Methods Cobalt-60 iso
Page 77 and 78: Materials and Methods specific bind
Page 79 and 80: Materials and Methods h half of the
Page 81 and 82: Materials and Methods phenol and th
Page 83 and 84: Materials and Methods 7. Filter ste
Page 85 and 86: Materials and Methods viz. EXO-SAP
Page 88 and 89: Materials and Methods Table 2: List
Page 90 and 91: Materials and Methods SMAD6 AREG HM
Page 92 and 93: Materials and Methods 3.9 Effect of
Page 94 and 95: Materials and Methods fresh eppendo
Page 96 and 97: Materials and Methods Power SYBR Gr
Page 98 and 99: Results Objective 1 To generate EBV
Page 100 and 101: Results Fig. 8: Frequency of second
Page 102 and 103: Results Table 6: Demographics of he
Page 104 and 105: Results Fig. 10: Cell population do
Page 106 and 107: Results Fig. 12: Cell surface marke
Page 108 and 109: Results 4.1.5 Expression of ATM gen
Page 110 and 111: Results Objective 2 To compare the
Page 112 and 113: Results Fig. 17: Standardization of
Page 114 and 115: Results Fig. 19: Standardisation of
Page 116 and 117: Results The cells were incubated fu
Page 118 and 119: Results 4.2.3.1 Percent G2 delay af
Page 120 and 121: Results 4.2.3.2 Effect of BPDE expo
Page 122 and 123: Results Fig. 29: Comparison of perc
Page 126 and 127: Results normalization with baseline
Page 128 and 129: Results 4.2.5.2 Real time PCR valid
Page 130 and 131: Results homozygous wild-type, heter
Page 132 and 133: Results Fig. 36b: Electrophoresis o
Page 134 and 135: Results Fig. 37a: Representative el
Page 136 and 137: Results Table 13: Consolidated geno
Page 138 and 139: Results LCL Genes NAT2(1) NAT2(2) N
Page 140 and 141: Results Fig. 38: Distribution of Ge
Page 142 and 143: Results Fig. 39: Various combinatio
Page 144 and 145: Results Table 14: Genotype-phenotyp
Page 146 and 147: Discussion Introduction Squamous ce
Page 148 and 149: Discussion stimulated lymphocytes w
Page 150 and 151: Discussion H2AX is currently the mo
Page 152 and 153: Discussion or G2/M phase arrest. As
Page 154 and 155: Discussion differential expression
Page 156 and 157: Discussion DNA repair genes and MPN
Page 158 and 159: Chapter 6 Summary and Conclusions
Page 160 and 161: Summary and Conclusions Statistical
Page 162 and 163: Bibliography 1. Dikshit, R.; Gupta,
Page 164 and 165: Bibliography 43. Lei, D.; Sturgis,
Page 166 and 167: Bibliography 76. Fry, R. C.; Svenss
Page 168 and 169: Bibliography 115. Kote-Jarai, Z.; M
Page 170 and 171: Bibliography 149. Hashibe, M.; Bren
Page 172 and 173: Bibliography 183. Bergamaschi, D.;
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Bibliography 216. Bondy, M. L.; Wan
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Indian J Med Res 135, June 2012, pp
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822 INDIAN J MED RES, JUNE 2012 mar
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824 INDIAN J MED RES, JUNE 2012 (h)
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826 INDIAN J MED RES, JUNE 2012 Tab
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828 INDIAN J MED RES, JUNE 2012 lik
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CASE REPORT Eben L. Rosenthal, MD,
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FIGURE 2. Chromosomal breakage anal
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FA along with ataxia telangiectasia
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LIFE09200604007 Tabish - Homi Bhabha National Institute

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