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Title
CERTIFICATE
DECLARATION
Thanks and Tributes
CONTENTS
I. Introduction and Review of Literature
Introduction
Review of Literature
CHEMICAL CARCINOGENS
Alkylating agents
Aralkylating agents
Arylhydroxylamines
GENETIC ASPECTS OF CANCER
Oncogenes
Ras gene
Tumour suppressor genes
pRB
p53
CARCINOGEN METABOLISM
CARCINOGENESIS AS A MULTISTAGE PROCESS
Initiation
Promotion
Progression
Malignant conversion
OXIDATIVE STRESS IN CHEMICAL CARCINOGENESIS
FREE RADICALS IN CARCINOGENECITY
MODIFICATION OF CARCINOGENESIS BY ANTIOXIDANTS
CHEMOPREVENTION
Ideal qualities of chemopreventative agents
Target populations
CLASSES OF CEMOPREVENTIVE AGENTS
Inhibitors of Carcinogen Formation
Blocking Agents
Inhibitors of cytochrome P450 enzymes
Inducers of Cytochrome P450 enzymes
Inducers phase 11 Enzymes
Scavengers of Electrophiles and free Radicals
Suppressing Agents
Inhibiters of Polyamine Metabolism
Inducers of Terminal Cell Differentiation
Modulators of Signal Transduction
Modulators of Hormonal / Growth Factor Activity
Inhibitors of Oncogene Activity
Promoters of Intercellular Communication
Restorers of Immune Response
Inducers of Apoptosis
Correctors of DNA Methylation Imbalances
Inhibitors of Basement Membrane Degradation
Inhibitors of Arachidonic Acid Metabolism
Current Human Intervention Trials
Phytochemicals in Chemoprevention
CANCER TREATMENT
ANTICANCER DRUGS FROM PLANTS
DNA Topoisomerase Targeted Drugs
Topoisomerase I-inhibitors
Topoisomerase II inhibitors
Microtubule-Targeting Drugs
Scope of the present study
Picrorhiza kurroa
Picroliv
Phyllanthus amarus
Plate: Phyllanthus amarus
Plate: Emblica offiinalis
Emblica offiinalis
II. Materials and Methods
Drugs
Chemicals
Tumour Cell lines
Animals
Determination of tumour reducing activity of plant products
Ascites tumour model
Determination of solid tumour model
Determination of in vitro antioxidant activity
Superoxide scavenging activity
Hydroxyl radical scavenging activity
Lipid peroxidation inhibition assay
Topoisomerase I and II inhibitory assays
Determination of cdc25 tyrosine phosphatase inhibitory assay
Estimation of protein
Estimation of tissue lipid peroxide
Estimation of serum lipid peroxide
Estimation of tissue superoxide dismutase (SOD) activity
Estimation of tissue catalase activity
Estimation of tissue glutathione (GSH)
Estimation of cytosolic glutathione-S- transferase (GST) activity
Estimation of Aniline Hydroxylase
Estimation of serum glutamate-pyruvate transaminase (GPT) activity
Estimation of alkaline phosphatase (ALP) activity
Estimation of Υ-glutamyl transpeptidase (Υ-GT) activity in tissue
Estimation of serum Υ-glutamyl traspeptidase (Υ-GT) activity
Estimation of serum bilirubin.
Histopathological methods
Hematoxylin and Eosin (H and E) staining
AgNOR staining
III. Pharmacological activity of Emblica officinalis Polyphenol
Section 1. Anticarcinogenic activity of Emblica officinalis polyphenol
1. Introduction
2. Materials and Methods
Preparation of polyphenolic fraction
Determination of in vitro antioxidant activity of EOP
Determination of the effect EOP on hepatocarcinogenesis induced by NDEA
3. Results
Isolation of polyphenolic fraction
Antioxidant activity of EOP
Effect of EOP on the hepatocarcinogenesis induced by NDEA
4. Discussion
Section 2. Induction of Apoptosis in Mouse and Human Carcinoma cell Lines by Emblica officinalis
1. Introduction
2. Materials and methods
Induction of apoptosis
3. Results
4. Discussion
Plate 2. H & E staining of DLA and CeHa cells
(a) DLA cells
(b) CeHa cells
(c) DLA cells treated with Emblica officinalis extract 200 &ml, show membrane blebbing and chromatin condensation
(d) CeHa cells treated with Emblica officinalis extract 200 pg/ml shows chromatin condensation
(e) DLA cells treated with EOP 200 pg/ml shows membrane blebbing
(f) CeHa cells treated with EOP 200 pg/ml shows chromatin condensation
Section 3. Antiulcerogenic activity of Emblica officinalis
1. Introduction
2. Materials and methods
Drug preparation
Ethanol-induced gastric ulcer
Indomethacin and histamine-induced gastric ulcers
Analytical methods
Histopathology
3. Results
List of Plates
Plate 3. Morphology and histology of sat stomach.
4.Discussion
IV. Pharmacological activity of Picroliv
Section 1. Antioxidant and Antitumour activity of Picroliv
1. Introduction
2. Materials and methods
2.1. Determination of in vitro antioxidant activity of Picroliv
2.2. Determination of the effect of Picroliv on aniline hydroxylase
2.3. Determination of the effect of Picroliv on Topoisomerase I and II
2.4. Determination of the effect of Picroliv on cdc25 tyrosine phosphatase.
2.5. Determination of the effect of Picroliv on ascites tumour development induced by transplanted tumours
2.6. Determination of the effect of Picroliv on solid tumour development induced by transplanted tumours
3. Results
3.1. Antioxidant activity of Picroliv
3.2. Effect of Picroliv on aniline hydroxylase, topoisomerase I and II and cdc25 tyrosine phosphatase
3.3. Effect of Picroliv on ascites tumour development
3.4. Effect of Picroliv on solid tumour development
4. Discussion
List of Plates
Plate 4. Mouse showing transplanted solid tumours, sarcomas and papillomas
Section 2. Anticarcinogenic activity of Picroliv
Effect of Picroliv on Chemical Carcinogenesis
1. Introduction
2. Materials and methods
2.1 Determination of the effect of Picroliv administration on NDEA-induced hepatocarcinogenesis
2.2 Determination of the effect of Picroliv treatment on DMH-induced toxicity
2.3 Determination of the effect of Picroliv on sarcoma induced by 20-MC
2.4 Determination of the effect of Picroliv treatment on papilloma formation initiated by DMBA and croton oil
3. Results
3.1 Effect of Picroliv treatment on NDEA induced hepatocarcinogenesis
3.2 Effect of Picroliv treatment in rats administered with DMH
List of Plates
Plate 5. Gross MorphoIogy and Histopathology of rat liver
3.3. Effect of Picroliv on the development of sarcomas induced by 20-MC
List of Plates
Plate 6. Histology of liver and kidney of rats
3.4. Effect of Picroliv on papilloma formation induced by DMBA and croton oil
4. Discussion
Section 3. Radioprotective and Chemoprotective activity of Picroliv
1. Introduction
2. Materials and methods
2.1. Determination of the effect of Picroliv on mice irradiated with Cobalt 60
2.2. Determination of the effect of Picroliv on cyclophosphamide-induced toxicity
3. Results
3.1 Effect of Picroliv on mice irradiated with Cobalt 60
3.2 Effect of Picroiiv on cyclophosphamide-induced toxicity
4. Discussion
V. Antitumor and Anticarcinogenic activity of Phyllanthus amarus and its mechanism of action
1. Introduction
2. Materials and methods
2.1 Determination of the effect P. amarus on ascites tumours
2.2 Determination of the effect P. amarus on solid tumours
2.3 Determination of the effect of P. amarus extract administration during induction of hepatocarcinogenesis by NDEA
2.4 Determination of the effect of P. amarus extract administration after induction of HCC by NDEA
2.5 Aniline hydroxylase inhibitory assay
2.6 Topoisomerase I and II inhibitory assays
2.7 Determination of cdc25 tyrosine phosphatase and cdc2 kinase inhibitory assays
3. Results
3.1 Effect of P. amarus on ascites tumours
3.2 Effect P. amarus on solid tumours
3.4 Effect of simultaneous administration of P. amarus on NDEA induced hepatocarcinogenesis
3.5 The effect of P. amarus extract administration after induction of HCC by NDEA
List of Plates
Plate 7. Gross morphology and histogathology of rat livers
3.6 Effect of P. amarus on aniline hydroxylase
3.7 Effect of P. amarus on Topoisomerase I and II
3.7 The effect of P. amarus on cdc25 tyrosine phosphatase and cdc2 kinase
4. Discussion
VI. Chemoprevention of Chemical Carcinogenesis by a Polyherbal formulation
1. Introduction
2. Materials and Methods
Preparation of Cancare
Determination of the effect Cancare on hepatocarcinogenesis induced by NDEA
Determination of the effect of Cancare on sarcoma induced by 20-MC
3. Results
Effect of Cancare administration on NDEA induced hepatocarcinogenesis
Effect of Cancare administration on 20-MC induced sarcoma development
4. Discussion
VII. Summary and Conclusion
References
Publications