ADVANCING ORAL CANCER DIAGNOSIS VIA TGFΒ SIGNALING PATHWAY
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Keywords
TGFβRII, ELISA, Transformation, TGFβ, Col2a1 and Oral cancer
Abstract
Background: The risk to a person's life posed by oral cancer can be significantly reduced if it is identified and confirmed early in the course of the illness. As a regulator, transforming growth factor-β (TGFβ) is essential to the control of numerous biological processes. Oral cancer patients' salivary TGFβ levels increase. It might therefore be applied as a diagnostic biomarker. The objective of this research was to create a reporter system in yeast cells that can identify TGFβ, serving as a non-invasive means of screening for oral cancer. Methodology: Transformation of yeast cells was verified by restriction digestion after transforming growth factor beta receptor II (TGFβRII) and collagen type II alpha chain I (col2a1) (intronic region) carrying vector were added. Gel electrophoresis analysis was used to express, optimise, and establish the recombinant proteins. Additionally, saliva from patients with oral cancer was combined with recombinant yeast cells, and the results were evaluated using green fluorescence protein fluorescence microscopy and the enzyme-linked immunosorbent assay (ELISA). Results: It was discovered that there was a substantial difference in TGFβ expression (P value < 0.05) between yeast cells that had undergone transformation and those that had not. Oral cancer patients' saliva samples were tested for TGFβ using ELISA, and samples of yeast treated with saliva were tested for TGFβ using solid-phase sandwich ELISA. The yeast samples containing TGFβRII and Col2a1 expression vectors showed the highest fluorescence. Conclusion: In this work, a green fluorescent protein reporter system based on yeast was developed for molecular detection of early oral cancer via saliva. ELISA and fluorescence microscopy were used to optimise and detect TGFβRII protein expression in S. cerevisiae. Oral cancer can be diagnosed non-invasively with this co-transformed method.
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Sidra Hasnain
Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore - Pakistan
Sana Javaid Awan
Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore - Pakistan
Associate Professor, Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore – Pakistan
Sana Khurshid
Department of Molecular Biology, Virtual University of Pakistan, Lahore - Pakistan
Anam Farzand
Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore - Pakistan
Mubeen Shahzadi
Institute of Molecular Biology and Biotechnology, The University of Lahore, Lahore - Pakistan