A.K. Saxena, Shalini, V. Singh, P. Singh
Published: February 28, 2024
Genet. Mol. Res. 23(1): GMR19195
DOI: https://doi.org/10.4238/gmr19195
Cite this Article:
A.K. Saxena, Shalini, V. Singh, P. Singh (2024). Differential expression of the vimentin gene modulated by MTHFR C677T polymorphism in circulating tumor cells isolated from breast cancer patients. Genet. Mol. Res. 23(1): GMR19195. https://doi.org/10.4238/gmr19195
About the Authors
A.K. Saxena, Shalini, V. Singh, P. Singh
Corresponding Author
Email: draksaxena1@rediffmail.com
ABSTRACT
Globally, breast cancer a common disease affecting women. The epithelial cells lose their characteristic features and gain mesenchymal properties during disease progression. During transition, the intermediate filament changes status from keratin to cell-adhesive molecules. Vimentin, a glycoprotein constitutes an intermediate filament of mesenchymal cells to maintain tissue architecture. The role of a tumor suppressor gene (p53) and methylenetetrahydrofolatereductase (MTHFR C677T) polymorphism becomes essential to explore the mechanism of isoform expression of vimentin in circulating tumor cells (CTCs) of breast cancer patients. We examined the frequency of vimentin gene expression along with isoforms and their correlation with p53 and MTHFR C677 polymorphism in CTCs. Blood samples were collected from clinically diagnosed cases of breast cancer (n=38) and age matched controls. Short term lymphocyte cultures were prepared for isolation of CTCs. cDNA was synthesized followed by PCR. PCR products were separated with agarose gel electrophoresis. There was an additional 250bp band, besides a conventional band of 323bp, with differential expression in CTCs. Variable frequency of over-expression was observed in two different bands in 31.3% (323bp) and 18.8% in isoforms (250bp) of the CTCs. Similarly, down-regulation varying between 43.8% (323bp) and 25.05% (250bp), while 25% cases showed a mutation of 323bp in CTCs. Data was further analyzed to calculate an95% confidence interval with odds ratio varying from 10.85 to 0.62 and 3.95 to 0.76 for over and under expression, respectively showing significant differences with respect to controls. MTHFR polymorphism showed higher (Tm) values (6.36) in the cases, where the mutation was observed as compared to those cases showing isoforms (5.59). The p53 gene mutation (273bp) appeared in nineteen out of thirty eight cases of breast cancer patients with a positive correlation between vimentin isoform expression and MTHFR polymorphism, suggesting a risk factor for this disease.
Key words: Breast cancer, Circulating tumor cell, Gene expression, Vimentin.