Expression Profiles of CDKN2A, MDM2, E2F2 and LTF Genes in Oral Squamous Cell Carcinoma

Abstract

Background: Oral squamous cell carcinoma (OSCC) is one of the most commonly detected neoplasms worldwide. Not all mechanisms associated with cell cycle disturbances are known in OSCC. Examples of genes involved in the control of the cell cycle are CDKN2A, MDM2, E2F2 and LTF. The aim of this study was to examine the possible association between CDKN2A, MDM2, E2F2 and LTF mRNA expression and influence on clinical variables. Methods: The study group consisted of 88 Polish patients. The gene expression levels were assessed by quantitative reverse transcription PCR. Results: We found no statistically significant differences in the expression level of CDKN2A, MDM2, E2F2 and LTF genes in tumour samples compared to margin samples. No association was found between the gene expression levels and clinical parameters, except E2F2. The patients with G2 tumours had a significantly higher gene expression level of E2F2 than patients with low-grade G1 tumours. Conclusions: We have not demonstrated that a change in expression profiles of genes has a significant impact on the pathogenesis of OSCC. It may also be useful to conduct further studies on the use of E2F2 expression profile changes as a factor to describe the invasiveness and dynamics of OSCC development.

miR-125b-5p, miR-155-3p, and miR-214-5p and Target E2F2 Gene in Oral Squamous Cell Carcinoma

“…The methodology for the extraction was presented in a previous study [ 76 ]. All tissue samples were homogenized with FastPrep ® -24 homogenizer (MP Biomedicals, Solon, CA, USA) with ceramic beads Lysing Matrix D (MP Biomedicals, Solon, CA, USA).…”

“…The RNA and miRNAs were extracted using the RNA isolation kit (catalog number RIK 001, BioVendor, Brno, Czech Republic) to the standard instruction. The concentration and purity of the isolated RNA was determined using spectrophotometry in NanoPhotometer Pearl UV/Vis Spectrophotometer (Implen, Munich, Germany) [ 76 ].…”

“…The methodology for the cDNA synthesis was presented in a previous study [ 76 ]. Total RNA (5 ng) was reverse-transcribed into cDNA using a high capacity cDNA reverse transcription kit with RNase inhibitor (Applied Biosystems, Foster City, CA, USA) according to manufacturer’s protocol.…”

“…The compound mix was prepared in a 20 μL volume containing: 2 μL of 10× Buffer RT; 0.8 μL of 25× dNTP mix (100 mM); 2 μL of 10× RT random primers; 1 μL of MultiScribe ® reverse transcriptase; 1 μL of RNase inhibitor; 3.2 μL of nuclease-free H 2 O; and 10 μL of previously isolated RNA. The reaction was processed in Mastercycler personal (Eppendorf, Hamburg, Germany) with the following thermal profile: 25 °C for 10 min, 37 °C for 120 min, 85 °C for 5 min, and 4 °C–∞ [ 76 ].…”

“…The qPCR was performed in a volume of 20 µL using 1 µL of cDNA, 10 µL of TaqMan ® fast advanced master mix (Applied Biosystems, Foster City, CA, USA), 1 µL of TaqMan ® gene expression assays (Assay ID: Hs00914334_m1 for E2F2 , and Assay ID: Hs03929097_g1 for GAPDH ), and 8 µL of nuclease-free H 2 O (EURx, Gdańsk, Poland). Thermal cycle for all analysed genes was: 95 °C for 20 s, followed by 40 cycles of 95 °C for 1 s, and 60 °C for 20 s [ 76 ].…”