Tesi di laurea magistrale
IDENTIFICAZIONE E CARATTERIZZAZIONE DI GENI IMPLICATI NEL MESOTELIOMA PLEURICO MALIGNO
Corso di studi
BIOLOGIA MOLECOLARE E CELLULARE
relatore Prof. Landi, Stefano
- gene silencing
- gene targeting
Data inizio appello
Data di rilascio
Introduction: Malignant Pleural Mesothelioma (MPM) is a rare and extremely aggressive cancer of the pleural cavity. It has been demonstrated the correlation between its onset and exposure to asbestos’s fibers. To date, it is very difficult to diagnose MPM at an early stage, given the non-specificity of the symptoms with which it occurs. The prognosis is poor, and an effective management strategy for this type of tumor is still absent. Therefore, the need for early diagnosis of this tumor and improvements in the prognosis, encourages the research and the identification of diagnostic and prognostic biomarkers specific and highly sensitive for MPM.
Previous works already done: This work is part of a larger project, aimed at the identification of genes, presenting a differential expression in tissues of healthy and pathological mesothelium. In this regard, a through literature research based on the transcriptomic’s works was carried out with the aim to collect all the genes deregulated in MPM. It was found that only some of these were cited as differentially expressed in no more than three studies. Given, the poor reproducibility among different works, an experimental validation of a total of 120 genes by Real-Time PCR was conducted by using an independent series MPM’s tissue and respective controls. Genes that have showed a positive result, were further validated in two cell lines of MPM. The validation has led to the individuation of 6 genes (CCNO, CFB, PDGFRB, SULF1, THBS2, TIMP3) that showed an “up-regulation”, statistically significant, both in tissues and cell lines.
Aim and methods: The project of this thesis had the aim to verify through RNA-interference technique, the potential role of TIMP3, PDGFRB e SULF1 in the carcinogenesis of MPM. After the key genes depletion, it will be then possible to observe how the malignant cells’ phenotype respond in terms of expression’s levels of transcripts (though Real Time PCR), proliferative capacity (through SRB assay), apoptosis (through the measuring of caspase activity), invasiveness capacity (through the Wound-Healing Assay), colony formation capacity (through Colony Formation Assay) and, finally, level of senescence (through the β-galactosidase’s measuring).
Preliminary results: From the first data SULF1’s silencing would seems to influence the proliferative and the colony formation’s capacity. This approach, thus, seems to be useful to understand the possible role played by genes in the healthy and in the malignant mesothelial tissue, and in the evaluating whether they could played a role either as possible markers of the disease or as potential therapeutic targets.
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