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  • TITLE
  • CERTIFICATE
  • DECLARATION
  • ACKNOWLEDGEMENT
  • DEDICATION
  • CONTENTS
  • LIST OF TABLES
  • LIST OF FIGURES
  • LIST OF ABBREVIATIONS
  • 1. INTRODUCTION AND REVIEW OF LITERATURE
  • 1.1 INTRODUCTION
  • 1.2 REVIEW OF LITERATURE
  • 1.2.1 Metastasis
  • 1.2.2 Metastatic cascade
  • 1.The metastatic cascade
  • 1.The pathogenesis of metastasis
  • 1.2.3 Growth rate and metastatic frequency
  • 1.2.4 Cell surface properties of metastatic tumour cells
  • 1.2.5 Cell attachment proteins in defining cell matrix
  • 1.2.5 (a) Attachment Proteins
  • 1.2.5 (b) Collagen
  • 1.2.5 (c) Proteoglycans
  • 1.2.6 Changes in matrix molecules and adhesion factors with cell transformation and malignancy
  • 1.2.7 Tumour cell detachment and transport
  • 1.2.8 Cell-cell and cell-matrix interactions during tumour metastasis
  • Tumour cell - endothelial cell interaction
  • Tumour cell-basement membrane interaction
  • Tumour cell - organ parenchyma cell interactions
  • 1.2.9 Tumour cell invasion mechanisms
  • Tumour cell - organ invasion
  • Turnour cell motility
  • Tumour cell chemotaxis
  • 1.2.10 Tumour cell growth mechanisms
  • Tumour autocrine growth factors
  • Tumour paracrine growth fnctors
  • 1.2.11 Anti tumour host response mechanisms
  • Types of antimetastatic host effector mechanism
  • Suppression of antimetastatic host response
  • 1.2.12 Basement membrane
  • Penetration by tumour cells
  • Basement membrane composition
  • 2.Three step hypothesis for tumour cell penetration of the basement membrane.
  • 1.2.13 Role of matrix degrading proteases in tumour invasion and metastasis
  • 1.2.13.1 Matrix Metalloproteinases (MMP)
  • 3 Some degradative enzymes and inhibitors synthesized by malignant and normal cells.
  • 2.Main members of the MMP family
  • Structure and Mechanism of Activation of MMPs
  • Role of proteinase Inhibitors in Invasion
  • 1.2.13.2 Serine Proteinases
  • 1.2.13.3 Cysteine proteinases
  • 1.2.13.4 Aspartyl proteinases
  • 1.2.14 Angiogenesis
  • 4.Different mechanisms involved in the switch to the angiogenic phenotype.
  • 3.Angiogenic and anti-angiogenic peptides
  • 1.2.15 Metastasis-Genetic control
  • 1.2.16 Role of polyphenolic compounds in cancer prevention
  • Flavonoids
  • 5 Structure of polyphenolic compounds.
  • Catechin and Epicatechin
  • Quercetin, Rutin and Morin
  • Naringin and Naringenin
  • Genistein and Daidzein
  • Phenolic acids
  • Curcumin
  • Ellagic Acid
  • Rasayanas
  • 1.2.17 Relevance of present study
  • 2. MATERIALS AND METHODS
  • 2.1 MATERIALS
  • 2.1.1 Test compounds
  • 2.1.2 Chemicals
  • 2.1.3 Reagents
  • 2.1.4 Instruments and devices
  • 2.15 Animals
  • 2.1.6 Tumour cell lines
  • 2.2 METHODOLOGY
  • 2.2.1 Sterilization of glasswares
  • 2.2.2 Preparation of culture media
  • 2.2.3 Maintenance of cell lines in tissue culture
  • 2.2.4 Maintenance of experimental animals
  • 2.2.5 Maintenance of tumours in animals
  • 2.2.6 Preparation of B 16F-10 melanoma cells for in vivo studies
  • 2.2.7 Preparation of B 16F-10 melanoma cells for in, vitro studies
  • 2.2.8 Determination of cell viability
  • 2.2.9 In vitro cytotoxicity studies
  • 2.2.10 Determination of the effect of compounds on the solidtumour development
  • 2.2.11 Determination of survival rate of tumour bearing animals
  • 2.2.12 Study on the in viuo invasion of B16F-10 melanoma cells inC57BW6 mice.
  • 2.2.13 Determination of the survival rate of metastatic tumourbearing animals.
  • 6. Lungs of metastatic tumour bearing C57BL/6 mice.
  • 2.2.14 Collagen matrix invasion assay
  • 2.2.15 Tumour cell adhesion experiment
  • 2.2.16 Tumour cell motility assay
  • 2.2.17 Gelatin zymography
  • Principle
  • Requirements
  • Procedure
  • 2.2.18 Estimation of protein
  • Principle
  • Reagents
  • Procedure
  • 2.2.19 Estimation of tissue hydroxyproline content
  • 2.2.20 Estimation of serum sialic acid
  • 2.2.21 Histopathological analysis
  • 2.2.22 Two Stage Carcinogenesis
  • 2.2.23 Methylcholanthrene induced chemical carcinogenesis
  • 2.2.24 Preparation of spleen cells
  • 2.2.25 Preparation of peritoneal macrophages
  • 2.2.26 Collection and preparation of SRBC
  • 2.2.27 Trypsinization of SRBC
  • 2.2.28 Determination of circulating antibody titre
  • 2.2.29 Determination of antibody forming cells
  • 2.2.30 Determination of macrophage mediated cytotoxicity
  • 2.2. 31 Statistical analysis
  • 3. CYTOTOXIC AND TUMOUR REDUCING ACTIVITY OF POLYPHENOLIC COMPOUNDS
  • 3.1 INTRODUCTION
  • 3.2 MATERIALS AND METHODS
  • 3.2.1 Test compounds
  • 3.2.2 Tumour cells
  • 3.2.3 In vitro short term cytotoxicity studies
  • 3.2.4 Cytotoxicity of polyphenolic compounds in tissue culture
  • 3.2.5 Effect of polyphenolic compounds on solid tumour development
  • 3.3 RESULTS
  • 3.3.1 In vitro cytot.oxic activity of polyphenolic compounds
  • 4.Short term in vitro cytotoxicity of polyphenolic compounds towards B16F-10 melanoma cells.
  • 5.Short term in vitro cytotoxicity of polyphenoliccompounds towards DLA cells
  • 3.3.2 Cytotoxicity of polyphenolic compounds in tissue culture
  • 6.Short term in vitro cytotoxicity of polyphenolic compounds towards EAT cells
  • 7.Long term in vitro cytotoxicity of polyphenolic compounds towards B16F-10 melanoma cells.
  • 8.Long term in vitro cytotoxicity of polyphenolic compounds towards L929 cells.
  • 7. Effect of polyphenolic compounds on B16F-10 melanoma induced solid tumour development.
  • 3.3.3 Effect of polyphenolic compounds on solid tumour development
  • 3.4 DISCUSSION
  • 9.Rate of survival of solid tumour (B16F-10) hearing animals treated with polyphenolic compounds.
  • 4. EFFECT OF POLYPHENOLIC COMPOUNDS ON LUNG METASTASIS OF B 16F- l0 MELANOMA CELLS IN MICE
  • 4.1 INTRODUCTION
  • 4.2 MATERIALS AND METHODS
  • 4.2.1 Animals
  • 4.2.2 Test compounds
  • 4.2.3 Effect of polyphenolic compounds on the inhibition of lung nodule formation
  • 4.2.4 Statistical analysis
  • 4.3 RESULTS
  • 4.3.1 Effect of polyphenolic compounds on the inhibition of lung metastasis
  • 10.Effect of polyphenolic compounds on the lung tumour nodule inhibition and survivalof metastatic tumour bearing animals.
  • 11.Effect of isoflavones on the lung tumour nodule inhibition and survival of metastatic tumour bearing animals.
  • 4.3.2 Effect of polyphenolic compounds on the survival rate of tumour bearing animals
  • 4.3.3 Effect of polyphenolic compounds on lung collagen hydroxyproline content
  • 12.Effect of polyphenolic compounds on the lung hydroxyproline and serum sialic acid content of metastatic tumour bearing animals.
  • 4.3.4 Effect of polyphenolic compounds on serum sialic acid level
  • 13.Effect of isoflavones on the lung hydroxyproline and serum sialic acid content of metastasic tumour bearing animals.
  • 4.3.5 Histopathological analysis of lungs of metastatic tumour bearing animals
  • 4.4 DISCUSSION
  • 8 Histopathological analysis of lungs of metastatic tumour bearing animals treated with polyphenolic compounds.
  • 5. EFFECT OF POLYPHENOLIC COMPOUNDS ON THE IN VITRO INVASION OF B 16F-10 MELANOMA CELLS THROUGH COLLAGEN MATRIX
  • 5.1 INTRODUCTION
  • 5.2 MATERIALS AND METHODS
  • 5.2.1 Cell line
  • 5.2.2 Polyphenolic compounds
  • 5.2.3 Effect of polyphenolic compounds on the collagen matrix invasion assay
  • 5.2.4 Effect of polyphenolic compounds on gelatin substrate gel electrophoresis
  • 5.2.5 Effect of polyphenolic compounds on tumour cell adhesion to collagen matrix
  • 5.2.6 Effect of polyphenolic compounds on tumour cell motility
  • 5.3 RESULTS
  • 5.3.1 Effect of polyphenolic compounds on collagen matrix invasion assay
  • 14.Effect of polyphenolic compounds on the inhibition of B 16F- 10 melanoma cells across collagen matrix.
  • 9. Effect of polyphenolic compounds on invasion of B 16F- 10 melanoma cells through collagen matrix
  • 5.3.2 Effect of polyphenolic compounds on gelatin substrate gel electrophoresis
  • 10 Zymographic analysis of B 16F-10 melanoma cell lysate.
  • 5.3.3 Effect of polyphenolic compounds on adhesion of B16F-10 cells to collagen matrix
  • 5.2.4 Effect of polyphenolic compounds on B 16F-10 cell motility
  • 5.4 DISCUSSION
  • 6. STUDIES ON ANTICARCINOGENIC ACTIVITY OF POLYPHENOLIC COMPOUNDS IN MICE
  • 6.1 INTRODUCTION
  • 6.2 MATERIALS AND METHODS
  • 6.2.1 Animal
  • 6.2.2 Compounds
  • 6.2.3 Effect of polyphenolic compounds on methylcholanthrene induced chemical carcinogenesis
  • 6.2.4 Effect of polyphenolic compounds on two stage carcinogenesis
  • 6.3 RESULTS
  • 6.3.1 Effect of polyphenolic compounds on the development of sarcoma induced by methylcholanthrene
  • 15.Effect of polyphenolic compounds on
  • 6.3.2 Effect of polyphenolic compounds on the survival of sarcoma bearing animals
  • 6.3.3 Effect of polyphenolic compounds on two stage carcinogenesis
  • 16. Effect of polyphenolic compounds on survival of.sarcoma bearing animals.
  • 17 Effect of polyphenolic compounds on Two Stage Carcinogenesis
  • 11. Effect of polyphenolic compounds on Two Stage Carcinogenesis.
  • 6.3.4 Effect of polyphenolic compounds on the collagen deposition during two stage carcinogenesis
  • 6.4 DISCUSSION
  • 18. Effect of polyphenolic compounds on the onset of Papilloma during Two Stage Carcinogenesis.
  • 19.Effect of polyphenolic compounds on skin collagen hydroxyproline content during carcinogenesis.
  • 7. IMMUNOLOGICAL MECHANISM OF ACTION OF POLYPHENOLIC COMPOUNDS IN MICE
  • 7.1 INTRODUCTION
  • 7.2 MATERIALS AND METHODS
  • 7.2.1 Test Compounds
  • 7.2.2 Animals
  • 7.2.3 Determination of circulating antibody titre
  • 7.2.4 Determination of antibody producing cells
  • 7.2.5 Effect of polyphenolic compounds on the production of TNF- α by macrophages
  • (a) In vivo activation of macrophages
  • (b) In vitro activation of macrophages
  • 7.2.6 Effect of polyphenolic compounds on the serum TNF- α levels in metastatic tumour bearing animals
  • 7.3 RESULTS
  • 7.3.1 Effect of polyphenolic compounds on circulating antibody titre
  • 7.3.2 Effect of polyphenolic compounds on antibody producing cells
  • 7.3.3 Effect of polyphenolic compounds on macrophage activation
  • 7.3.4 Effect of polyphenolic compounds on the serum TNF- α production in metastatic tumour bearing animals
  • 20.Effect of polyphenolic compounds on the circulating antibody titre in mice.
  • 21.Effect of polyphenolic compounds on antibody producing cells.
  • 7.4 DISCUSSION
  • 12 Effect of polyphenolic compounds on the serum TNF-α level in metastatic tumour bearing animals.
  • 8. STUDIES ON THE ANTIMETASTATIC ACTIVITIES OF ANTIINFLAMMATORY AGENTS IN MICE
  • 8.1 INTRODUCTION
  • 8.2 MATERIALS AND METHODS
  • 8.2.1 Animals
  • 8.2.2 Cell line
  • 8.2.3 Test compounds
  • 8.2.4 Effect of antiinflammatory agents in the inhibition of lung tumour nodule formation
  • 8.2.5 Statistical analysis
  • 8.3 RESULTS
  • 8.3.1 Effect of antiinflammatory agents on the inhibition of lung metastasis
  • 22.Effect of antiinflammatory agents on BI6F-10 melanoma induced lung metastasis.
  • 8.3.2 Effect of anti inflammatory agents on the survival rate of metastatic tumour bearing animals
  • 8.3.3 Effect of antiinflammatory agents on the lung collagen hydroxy proline content
  • 8.3.4 Effect of antiinflammatory agents on serum sialic acid level
  • 23.Effect of antiinflammatory agents on the rate of survival of metastasic tumour bearing animals.
  • 24.Effect of anti inflammatory agents on the serum sialic acid and lung hydroxyproline content of metastatic tumour bearing animals.
  • 8.3.5 Histopathological analysis
  • 8.4 DISCUSSION
  • 9. ANTICARCINOGENIC AND ANTIMETASTATIC ACTIVITY OF RASAYANAS
  • 9.1 INTRODUCTION
  • 9.2 MATERIALS AND METHODS
  • 9.2.1 Animals
  • 9.2.2 Cell line
  • 9.2.3 Materials
  • 9.2.4 Determination of the effect of Rasayanas in the inhibition of lung tumour nodule formation and survival
  • 9.2.5 Effect of Brahma Rasayana on the development of methylcholanthrene induced chemical carcinogenesis
  • 9.3 RESULTS
  • 9.3.1 Effect of Rasayanas on the inhibition of lung tumour nodule formation
  • 9.3.2 Effect of Rasayanas on the survival of metastatic tumour bearing animals
  • 25.Effect of various Rasayanas on the inhibition of lung tumour nodules.
  • 26.Effect of various Rasayanas on the survival of metastatic tumour bearing animals.
  • 9.3.3 Effect of Rasayanas on the lung collagen hydroxyproline content
  • 9.3.4 Effect of Rasayanas on the serum sialic acid level
  • 27.Effect of various Rasayanas on the serum sialic acid and lung hydroxyproline content of metastic tumour bearing animals.
  • 9.3.5 Histopathological analysis
  • 9.3.6 Effect of Brahma Rasayana administration on sarcoma development
  • 9.3.7 Effect of Brahma Rasayana treatment on the survival of animal injected with methyl cholanthrene
  • 13. Histopathological analysis of lungs of metastatic tumour bearing animals treated with Rasayanas.
  • 28.Effect of Brahma Rasayana on the development of sarcomα
  • 29.Effect of Brahma Rasayana on the survival of animals induced with methylcholanthrene.
  • 9.4 DISCUSSION
  • 10. SUMMARY AND CONCLUSION
  • BIBLIOGRAPHY
  • LIST OF PAPERS