scholarly journals Comparative Evaluation on the Expression of Pi3 Kinase - AKT- mTOR Signalling Molecules in Oral Squamous Cell Carcinoma - A Real Time PCR Based Approach

Author(s):  
Krishnapriya Umashankar ◽  
J. Selvaraj ◽  
Pratibha Ramani

Background: Oral squamous cell cancer (OSCC) develops as a result of the accumulation of many genetic mutations that are influenced by genetic predisposition. Upon acquisition of genetic predisposition, the precancerous cells will transform into malignant cells culminating into carcinomas. Advances in genetic research over the past few decades have rendered early detection possible. Aim: To compare the gene expression of Pi3 Kinase, AKT and mTOR in OSCC and to correlate the expression levels of these molecules with the survival in OSCC patients. Also to understand the role of Pi3 Kinase pathway in OSCC progression thereby attempting targeted therapy in OSCC patients. Materials and Methods: 10 OSCC samples as well as normal healthy samples were collected and RNA isolation was done using RNA easy kit from Qiagen (Valencia, CA), and then subjected to cDNA synthesis using Human TGF-β1, Human GSK-3β and Human Pi3 kinase primers. Real time PCR was performed using gene specific primers at 40 cycles. The results were retrieved, tabulated and analyzed. Results: The current research results revealed that there were up regulation of mRNA expression in The PI3K/AKT/mTOR in OSCC patients than in healthy individuals. On comparison, mTOR showed highest mRNA expression levels than AKT and PI3K. Conclusion: Overexpression of Pi3 kinase, AKT, mTOR plays a crucial role in progression of oral cancer and targeting Pi3 kinase/mTOR pathways could be a novel and targeted approach for OSCC.

Author(s):  
Krishnapriya Umashankar ◽  
J. Selvaraj ◽  
Pratibha Ramani

Background: Oral Squamous Cell Carcinoma (OSCC) is the most common malignancies which accounts for 90% of oral cancer worldwide. The 5 year survival rate of OSCC is not more than 60% due to tumor metastasis and subsequent recurrence. Aim: To compare the gene expression of ITG β-1, MMP9 and Vimentin in OSCC tissue samples and normal tissue samples and to correlate the expression levels of these molecules with the pathological grading and survival in OSCC patients. This would facilitate the understanding of EMT in OSCC progression thereby targeting this pathway for treatment of OSCC patients in near future. Materials and Methods: 10 OSCC samples as well as normal healthy samples were collected and RNA isolation was done using TRIR kit, and then subjected to cDNA synthesis using ITG β-1, MMP9 and Vimentin primers. Real time PCR was performed using gene specific primers at 40 cycles. The results were retrieved, tabulated and analyzed. Results: The current research results revealed that there were up regulation of mRNA expression in ITG β-1, MMP9 and Vimentin in OSCC patients than in healthy individuals. On comparison, MMP9 showed highest mRNA expression levels than ITG β-1 and Vimentin Conclusion: Over expression of ITG B-1, MMP9, Vimentin plays a crucial role in progression of oral cancer and targeting EMT molecules could be an effective targeted approach for OSCC.


2006 ◽  
Vol 21 (1) ◽  
pp. 30-39 ◽  
Author(s):  
M. Labuhn ◽  
V. Vuaroqueaux ◽  
F. Fina ◽  
A. Schaller ◽  
I. Nanni-Metellus ◽  
...  

The assessment of ERα, PgR and HER2 status is routinely performed today to determine the endocrine responsiveness of breast cancer samples. Such determination is usually accomplished by means of immunohistochemistry and in case of HER2 amplification by means of fluorescent in situ hybridization (FISH). The analysis of these markers can be improved by simultaneous measurements using quantitative real-time PCR (Qrt-PCR). In this study we compared Qrt-PCR results for the assessment of mRNA levels of ERα, PgR, and the members of the human epidermal growth factor receptor family, HER1, HER2, HER3 and HER4. The results were obtained in two independent laboratories using two different methods, SYBR Green I and TaqMan probes, and different primers. By linear regression we demonstrated a good concordance for all six markers. The quantitative mRNA expression levels of ERα, PgR and HER2 also strongly correlated with the respective quantitative protein expression levels prospectively detected by EIA in both laboratories. In addition, HER2 mRNA expression levels correlated well with gene amplification detected by FISH in the same biopsies. Our results indicate that both Qrt-PCR methods were robust and sensitive tools for routine diagnostics and consistent with standard methodologies. The developed simultaneous assessment of several biomarkers is fast and labor effective and allows optimization of the clinical decision-making process in breast cancer tissue and/or core biopsies.


2002 ◽  
Vol 67 (2) ◽  
pp. 225-234 ◽  
Author(s):  
Julieta Alfonso ◽  
Guido D. Pollevick ◽  
Anja Castensson ◽  
Elena Jazin ◽  
Alberto C.C. Frasch

2020 ◽  
Vol 98 (Supplement_2) ◽  
pp. 35-35
Author(s):  
Maegan A Reeves ◽  
Courtney E Charlton ◽  
Terry D Brandebourg

Abstract Given adipose tissue is histologically classified as connective tissue, we hypothesized expression of extracellular matrix (ECM) components are significantly altered during adipogenesis. However, little is known about the regulation of the ECM during adipose tissue development in the pig. Therefore, the objective of this study was to characterize expression of ECM components during porcine adipogenesis. Primary cultures of adipose tissue stromal-vascular cells were harvested from 3-day-old neonatal pigs (n=6) and preadipocytes induced to differentiate in vitro for 8 days in the presence of insulin, hydrocortisone, and rosiglitazone. Total RNA was extracted from these cultures on days 0 and 8 post-induction. Real-time PCR was then utilized to determine changes in mRNA expression for collagen type I alpha 1 chain (COL1A), collagen type I alpha 2 chain (COL2A), collagen type I alpha 3 chain (COL3A), collagen type I alpha 4 chain (COL4A), collagen type I alpha 6 chain (COL6A), biglycan, fibronectin, laminin, nitogen-1 (NID1), matrix metallopeptidase 2 (MMP2), matrix metallopeptidase 9 (MMP9), metallopeptidase inhibitor 3 (TIMP3). The mRNA abundances of COL1A, COL3A and MMP2 were significantly downregulated 2.86-fold (P < 0.05), 16.7-fold (P < 0.01) and 3.1-fold (P < 0.05) respectively in day 8 (differentiated) compared to day 0 (undifferentiated) cultures. Meanwhile, mRNA abundances were significantly upregulated during adipogenesis for the COL2A (2.82-fold; P < 0.05), COL4A (2.01-fold; P < 0.05), COL6A (2.8-fold; P < 0.05), biglycan (49.9- fold; P < 0.001), fibronectin (452-fold; P < 0.001), laminin (6.1-fold; P < 0.05), NID1(47.4-fold; P < 0.01), MMP9 (76.8- fold; P < 0.01), and TIMP3(3.04-fold; P < 0.05) genes. These data support the hypothesis that significant changes in ECM components occur during porcine adipogenesis. Modulating adipose tissue ECM remodeling might be a novel strategy to manipulate adiposity in the pig.


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