A Review on Genotoxicity

 

Ahamed Noor Mansoori, Rupesh K. Gautam*, Prafulla C. Tiwari

Department of Pharmacology, Jaipur College of Pharmacy, ISI-15, RIICO Institutional Area, Tonk Road, Sitapura, Jaipur, India-302022

*Corresponding Author E-mail: rgautam3906@yahoo.co.in

 

ABSTRACT:

Genotoxicity studies used to investigate the potency of a compound to interact with the constitution of genetic. An alteration of the genetic code via gene mutations (point mutations) or DNA strand breaks may lead to fundamental changes DNA damage in a somatic cell may result in a somatic mutation, which may lead to malignant transformation (cancer). A genotoxin is an agent or chemical that can cause DNA or chromosomal damage. Such damage in a germ cell has the potential to cause a heritable altered trait (germline mutation). Many chemical carcinogens/ mutagens undergo metabolic activation to reactive species that bind covalently to DNA, and the DNA adducts thus formed can be detected in cells and in human tissues by a variety of sensitive techniques. The present study was aimed to provide information regarding genotoxicity.

 

KEY WORDS: Genotoxin, Chromosomal damage, Carcinogen, Gene mutations.

 


 

INTRODUCTION:

Genotoxicity is an important word in genetics defined as a destructive effect on a cell's genetic material (RNA, DNA) affecting its integrity. Genotoxins are mutagens; they can cause mutations. Genotoxins include both chemical and radiation genotoxins. A substance that has the property of genotoxicity is known as a genotoxin. 1

 

Genotoxicity describes the property of chemical agents that destroy the genetic information within a cell causing mutations, which may lead to cancer. While genotoxicity is often confused with mutagenicity, it is important to know that all mutagens are genotoxic; however, not all genotoxic substances are mutagenic. 2

Genotoxin: A genotoxins is a chemical or agent that can cause chromosomal or DNA damage. Such damage in a germ cell has the potential to cause a heritable altered trait (germ line mutation). DNA damage in a somatic cell may result in a somatic mutation, which may lead to malignant transformation (cancer). Many in-vitro and in-vivo tests for genotoxicity have been developed that, with a range of endpoints, detect DNA damage or its biological consequences in prokaryotic (e.g. bacterial) or eukaryotic (e.g. mammalian, yeast or avian) cells.

 

The detection and characterization of DNA adducts in human tissues gives clues to the etiology of human cancer. Characterizations of gene mutations in human tumor’s, in common with the known mutagenic profiles of genotoxins in experimental systems, may provide further insight into the role of environmental mutagens in human cancer. 3

 

Agents capable of causing direct or indirect damage to DNA:

·      Electrophilic species forming covalent adducts to DNA e.g. Alkylating agents –Aryl nitrenium ions, Diol epoxides of PAH etc.

·      Nucleoside analogues

·      UV and ionizing radiations

·      Topoisomerase inhibitors

·      Reactive oxygen species

·      Protein synthesis inhibitors 1

·      Herbal Plants like Aconite, Alfa alfa, Calamus, Borage, Chaparrel,  Colts foot, Comfrey, Germander, Ephedra, Senna, Ginkgo biloba, Ginseng, Glycyrrhiza glabra (Liquorice), Isabghol, Aloe vera, Sassafras, Silybum marianum 4.

 

Figure 1: The inter-relationships of genotoxicity and carcinogenicity

 

Herman Druckrey, at a conference in Sweden, first used the word genotoxic for chemicals that can react with DNA, and thus have the potential of being mutagenic, carcinogenic and cell transforming. It has been popularized the use of the term genotoxic as agents that were DNA-reactive, directly or after biochemical activation, with appropriate fractions from liver or other tissue of rodents or humans. In sharp contrast, there are other chemicals and agents that are clearly not mutagenic, but that have the ability of increasing the efficiency or effectiveness of a genotoxic carcinogen. The classic promoters of carcinogenesis fall into that class, but there are other non genotoxic agents such as immunological elements, endocrine factors and related properties that are of demonstrated relevance in the complex overall carcinogenic process, as we now understand. One key property of this class of agents is that they may control the rate of DNA synthesis and of cell division. This is important ever since Cleaver discovered repair of DNA. A cell containing mutated DNA yield an abnormal fraudulent DNA only after such a DNA has served as a template in the synthesis of new DNA during mitosis and cell cycling. Thus, the rate of cell division is an important parameter. Nevertheless, simple screening bioassays can play very important role in pre-regulatory genotoxicity screening strategies because of their rapid genotoxicity detection requirement that is demanded in preliminary decision making process. 5

 

The alteration can have direct or indirect effects on the DNA:

The induction of mutations, mistimed event activation, and direct DNA damage leading to mutations. The permanent, hereditary changes can affect either somatic cells of the organism or germ cells to be passed on to future generations. Cells prevent expression of the genotoxic mutation by either DNA repair or apoptosis; however, the damage may not always be fixed leading to mutagenesis.

 

Mechanisms:

The genotoxic substances induce damage to the genetic material in the cells through interactions with the DNA structure and sequence. For example, the transition metal chromium interacts with DNA in its high-valent oxidation state so to occur DNA lesions leading to carcinogenesis. The metastable oxidation state Cr (V) is achieved through reductive activation. The researchers performed an experiment to study the interaction between DNA with the carcinogenic chromium by using a Cr (V)-Salen complex at the specific state of oxidation. The interaction was specific to the guanine nucleotide in the genetic sequence.

 

Figure: 2 Definition of transitions and transversions, they are a common mutation caused by genotoxic compounds

 

In order to narrow the interaction between the Cr (V)-Salen complex with the guanine base, the researchers modified the bases to 8-oxo-G so to have site specific oxidation. The reaction between the two molecules caused DNA lesions; the two lesions observed at the modified base site were spiroiminodihydantoin and guanidinohydantoin. To further analysis of the site of lesion, it was observed that polymerase stopped at the site and adenine was inappropriately incorporated into the DNA sequence opposite of the 8-oxo-G base. Therefore, these lesions predominately have G >T transversions. High-valent chromium is seen to act as a carcinogen as researchers found that "the mechanism of damage and base oxidation products for the interaction between DNA and high-valent chromium are relevant to in-vivo formation of DNA damage leading to cancer in chromate-exposed human populations." Consequently, it shows how high-valent chromium can act as a carcinogen with 8-oxo-G forming xenobiotics

. 2

 


 

Mode of Action:

 

Figure 3: Mode of action 6

 


The following standard test battery is recommended based upon the considerations mentioned above:

i)         A test for gene mutation in bacteria.

ii)       An in-vitro test with cytogenetic evaluation of chromosomal damage with mammalian cells or an in-vitro mouse lymphoma assay.

iii)    An in-vivo test for chromosomal damage using rodent hematopoietic cells. 7

 

Purpose of genotoxicity:

·      To investigate the safety and efficacy of new chemical entities before their release into the market.

·      To detect the responsible for heritable effect on germ cells and impose risk to future generations.

·      To give quantitative estimate of the contribution of chemical agents to the incidence of genetic diseases and risk characterization for cancer.

·      To detect the hazard identification with respect to DNA damage in the form of structural chromosomal aberration, gene mutation, recombination and numerical changes. 9

 

Purpose of genotoxicity test assays

·      Assays allow detection of a drug’s potential for genotoxicity early in development of drug

·      Assays designed to be more sensitive to damage in order to enhance hazard identification.

·      Assays are inexpensive, have high statistical power, are generally reproducible and detect a variety of genotoxic end-points. 10

 

Genotoxic risk:

 

Figure 4: Genotoxic risk

 

OECD test guidelines for genotoxicity and mutagenicity testing 8

TG 471

Bacterial Reverse Mutation Test (Ames Test)

TG 472

Genetic Toxicology: Escherichia coli, reverse assay

TG 473

In-vitro Mammalian Chromosome Aberration Test

TG 474

Mammalian Erythrocyte Micronucleus Test

TG 475

Mammalian Bone Marrow Chromosome Aberration Test

TG 476

In-vitro Mammalian Cell Gene Mutation Test

TG 477

Genetic Toxicology: Sex-linked Recessive Lethal Test in Drosophila melanogaster

TG 478

Genetic Toxicology: Rodent Dominant Lethal Test

TG 479

Genetic Toxicology: In-vitro Sister Chromatid Exchange Assay in Mammalian Cells

TG 480

Genetic Toxicology: Saccharomyces cerevisiae, Gene Mutation Assay

TG 481

Genetic Toxicology: Saccharomyces cerevisiae, Mitotic Recombination Assay

TG 482

Genetic Toxicology: DNA Damage and Repair, Unscheduled DNA Synthesis in Mammalian Cells in-vitro

TG 483

Mammalian Spermatogonial Chromosome Aberration Test

TG 484

Genetic Toxicology: Mouse Spot Test

TG 485

Genetic Toxicology: Mouse Heritable Translocation Assay

TG 486

Unscheduled DNA Synthesis (UDS) Test with Mouse Liver Cells in-vitro

TG 487

In-vitro Mammalian Cell Micronucleus Test

 

Usage of genotoxicity tests:

·      In-vivo and in-vitro genotoxicity tests may be used to give information regarding the efficiency of the compound in directly or indirectly inducing mutations and an alert about the probability of carcinogenicity of the compound.

·      Determine toxicity profiles of chemicals

·      Reveal the molecular mechanism underlying the chemicals genotoxic and carcinogenic effects

·      Identify hazards in risk assessment within molecular epidemiologic research with regards  to occupational and environmental chemicals

·      Monitor the diseases and effectiveness of clinical treatments

·      Develop regulations concerning cosmetic, medical and industrial chemicals according to international and national guidelines. 11

 

CONCLUSION:

Genotoxicity describes the property of chemical agents that damages the genetic information within a cell causing mutations in genetics. This review will be fruitful those involved in the study and research on genotoxicity. It will become an extensive tool for providing information on genotoxicity.

 

REFERENCES:

1.     Shah SU. Importance of genotoxicity & S2a guidelines for genotoxicity testing for pharmaceuticals. Iosr Journal of Pharmacy and Biological Sciences. 1(2): 2012:43-54,

2.     Available from:URL : http://En.Wikipedia.Org/Wiki/Genotoxicity, accessed on May, 2014

3.     Phillips DH, Arlt VM. Genotoxicity: damage to DNA and its consequences. EXS. 2009; 99: 87-110.

4.     Philomena G. Concerns regarding the safety and toxicity of medicinal plants -An overview. Journal of Applied Pharmaceutical Science. 2011; 1(6): 40-44.

5.     Vasantha KC, Nagarathna PKM, Kulakarni SC, Sainadh NS. Evaluation of antimutagenic effect of flavonoid of Kigelia africana on Swiss-albino mice. International Journal of Pharmaceutical Sciences Review and Research 2013; 21(1): 105-8.

6.     Available from: URL: http://Openi.Nlm. Nih.Gov/Imgs/ Rescaled512/2759963_1471-2105-10-303-4.Png, accessed on May, 2014

7.     ICH Harmonised Tripartite Guideline S2B. Genotoxicity: A standard battery for genotoxicity testing of pharmaceuticals. 1998; 3-9.

8.     Available from URL http://www.alttox.org/ttrc/toxicity-tests/genotoxicity/references. html, accessed on April, 2014

9.     Savage JR. Classification and relationships of induced chromosomal structural changes. J Med Genetics. 1976; 13: 103-22

10.   Suzanne R. Thornton-J. International Research CMC Conference, Sanofi Aventis, U.S. 2006; 1-8

11.   Goncacak M, Mircigil C. Genotoxicity tests from biomarker studies to the regulations: National perspective. Journal Pharmaceutical Science. 2009; 34: 217–32

 

 

 

Received on 12.07.2014          Accepted on 16.08.2014        

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Asian J. Pharm. Res. 4(3): July-Sept. 2014; Page 162-165