Reverse Mutation Concept in Cancer Biology Term Paper

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Evaluating 2-Aminoanthracene as a Possibly Cancerogenic Compound

In the environment where animal testing is not allowed, determining possible genotoxicity of a compound is possible with the help of the Ames test (Samiei et al. 121). The specified instrument represents a “bacterial reverse mutation assay” (Samiei et al. 121), which will ostensibly help detect a mutagenic compound. The test involves using the strains of Salmonella/E.coli. As a rule, the following strains are used to detect the presence of cancerogenic elements: “TA97, TA98, TA100, TA102, TA104, TA1535, TA1537, and TA1538” (U.S. Department of Health and Human Services). The test is based on the concept of reverse (back) mutation. The bacteria are cultured in the presence of 2-Aminoanthracene as a supposedly mutagenic compound and without histidine. In case Salmonella/E.coli becomes capable of producing histidine and continues to proliferate, the mutagenic nature of 2-Aminoanthracene will be proven (Ruiz-Pérez et al. 6743). Since the test is known for delivering false-positive results, the control tools such as TA98, TA98 and TA1538, TA100 and TA1535, and TA102 will have to be utilized (U.S. Department of Health and Human Services).

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Human Oncogene in Fibroblasts: Cloning, Activity Analysis, and the Human Genome

To determine whether it is the human cellular oncogene that affects the development of the fibroblasts, one will have to consider using the DNA sequences known as Alu sequences as the key tool (Yang et al. 5812). Since Alu elements are typically viewed as the cause of human mutations, they will have to be used as the testing tool (Yang et al. 5812). The DNA sequence from the mouse cells will be isolated so that it could be cloned into the bacteriophage (Sherif and Broome 124). Using the transposon tagging framework, which implies tagging the genes that do not coincide with the regular process of gene expression, one will be able to detect the presence of the human cellular oncogene in the fibroblasts developed by the mice (Zhang et al. 3134). The hybridization of phage and the Alu probe will point to the presence of the human mutagen in the cell (Yang et al. 5813).

Defining the Possibility of Ras Being Responsible for the Cell Transformation

To test whether the phenotype was transformed by Ras, one will have to consider using a signaling inhibitor that will allow detecting the possibility of Ras being responsible for the malignant transformations in cells. Particularly, the application of the KIF2C protein will have to be considered to identify the ostensible connection between Ras and the transformation of cells. To be more specific, the use of KIF2C as the kinesin family member will allow inhibiting the activation of ERK1/2, which, in turn, spurs the mTORC1 activity in a mutated gene (Zaganjor 10568). Therefore, in case the activity mentioned above is observed in the mutant gene, the fact that it was Ras that triggered the transformation can be considered proven.

Mechanistic Role of Rb and Protein Mutations: What Produces the Transformed Phenotype

The Rb mechanism causes alterations in chromatin structures of cells (Scott et al. 28972). The abnormal expression of the gene may lead to cancer cells developing the ability to either activate or repress the process of gene transcription (Ferraro 196). As a result, the genomes of cancer cells can be reprogrammed so that the development of malignant elements could be stopped, activated, or aggravated (Lin and Zhao 388). The synonymous mutations that occur in the process include “G12, G13, and G60” (Waters e0163272).

Works Cited

Ferraro, Angelo. “Altered Primary Chromatin Structures and Their Implications in Cancer Development.” Cellular Oncology, vol. 39, no. 3, 2016, pp. 195-210. Web.

Lin, Run-Long, and Lu-Jun Zhao. “Mechanistic Basis and Clinical Relevance of the Role of Transforming Growth Factor-β in Cancer.” Cancer Biology & Medicine, vol. 12, no. 4, 2015, pp. 385-393. Web.

Ruiz-Pérez, Nancy J., et al. “Mutagenic and Antimutagenic Effects of Heterotheca Inuloides.” Scientific Reports, vol. 4, no. 1, 2014, pp. 6743-6748. Web.

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Samiei, Mohammad, et al. “Investigating the Mutagenic Effects of Three Commonly Used Pulpotomy Agents Using the Ames Test.” Advanced Pharmaceutical Bulletin, vol. 5, no. 1, 2015, pp. 121-125. Web.

Scott, Milcah C., et al. “Aberrant Retinoblastoma (RB)-E2F Transcriptional Regulation Defines Molecular Phenotypes of Osteosarcoma.” Journal of Biological Chemistry, vol. 290, no. 47, 2015, pp. 28070-28083. Web.

U.S. Department of Health and Human Services. “The Salmonella/E. coli Mutagenicity Test or Ames Test.” National Toxicity Program, Web.

Waters, Andrew M., et al. “Single Synonymous Mutations in KRAS Cause Transformed Phenotypes in NIH3T3 Cells.” PloS One, vol. 11, no. 9, 2016, p. e0163272. Web.

Yang, Ruifeng, et al. “Direct Conversion of Mouse and Human Fibroblasts to Functional Melanocytes by Defined Factors.” Nature Communications, vol. 5, pp. 5807-5832. Web.

Zaganjor, Elma, et al. “Ras Transformation Uncouples the Kinesin-Coordinated Cellular Nutrient Response.” Proceedings of the National Academy of Sciences, vol. 111, no. 29, 2014, pp. 10568-10573. Web.

Sherif, Zaki A Carolyn W Broome. “Restoration of Mitochondrial Gene Expression Using a Cloned Human Gene in Chinese Hamster Lung Cell Mutant.” Advanced Techniques in Biology & Medicine, vol. 3, no. 1, 2015, pp. 120-146. Web.

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Zhang, Hengzhu, et al. “Reprogramming A375 Cells to Induced‑Resembled Neuronal Cells by Structured Overexpression of Specific Transcription Genes.” Molecular medicine reports, vol. 14, no. 4, 2016, pp. 3134-3144. Web.

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