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Genetic analysis of oral squamous cell carcinoma by cDNA microarrays focused apoptotic pathway
L Lo Muzio1, A Santarelli, M Emanuelli
1Dipartimento di Scienze Chirurgiche, Università di Foggia, Italy. lomuziol@tin.it
Abstract:
We investigated mRNA expression of the genes involved in the apoptotic mechanism in oral squamous cell carcinoma (OSCC) by cDNA microarray. The aim of this study was to identify genes mainly involved in tumorigenesis, comparing the difference of gene expression in neoplastic and non-neoplastic tissues. Eight frozen samples of OSCC and the corresponding normal oral mucosa were treated to obtain mRNA. The mRNA extracted from these specimens was converted into cDNA and analyzed with SuperArray GEArray Q Series Human Apoptosis Gene Array kit. Our results showed that in OSCC there is a different expression of CRADD, FADD, ATM and APAF-1 genes compared to normal mucosa. Real-Time PCR, and Western blot analysis were performed on a separate cohort of patients in order to confirm the results obtained by DNA microarray. Our analysis of apoptotic process through microarray technology confirmed that different molecules could be responsible or favour the imbalance of apoptosis in cancer tissues. Microarray technology has made it possible to analyze the expression of multiple genes in a single experiment. However, most commercial array kits, designed to include as many genes as possible, produce a vast amount of data that often is difficult to interpret. In addition, the cost of equipment is often prohibitive. In contrast, the focused kit used was a complete, affordable and effective method to improve knowledge of molecular specific pathways.
Insights
This study explored gene expression in oral squamous cell carcinoma (OSCC), finding differences in apoptosis-related genes like CRADD and FADD between cancer and normal tissues.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Oral squamous cell carcinoma (OSCC) is a prevalent cancer.
- Understanding the molecular mechanisms, particularly apoptosis, is crucial for OSCC tumorigenesis.
- Gene expression profiling offers insights into these mechanisms.
Purpose of the Study:
- To investigate mRNA expression of apoptosis-related genes in OSCC.
- To identify genes involved in OSCC tumorigenesis by comparing neoplastic and non-neoplastic tissues.
- To assess the utility of focused microarray kits for studying specific molecular pathways.
Main Methods:
- cDNA microarray analysis using SuperArray GEArray Q Series Human Apoptosis Gene Array kit.
- Extraction and analysis of mRNA from 8 OSCC and corresponding normal oral mucosa samples.
- Validation of key gene expression findings using Real-Time PCR and Western blot analysis.
Main Results:
- Significant differences in the expression of CRADD, FADD, ATM, and APAF-1 genes were observed in OSCC compared to normal oral mucosa.
- Microarray analysis revealed distinct molecular players potentially contributing to apoptosis imbalance in OSCC.
- Focused apoptosis gene array provided an affordable and effective method for pathway analysis.
Conclusions:
- Gene expression analysis highlights specific apoptosis-related genes (CRADD, FADD, ATM, APAF-1) differentially expressed in OSCC.
- These molecular differences may contribute to the development and progression of oral squamous cell carcinoma.
- Focused microarray kits are valuable tools for in-depth study of specific molecular pathways in cancer research.
