The regulatory role of non-coding RNAs (ncRNAs) and their implication in tumor progression have only occurred in recent decades. To date, among the most studied classes of ncRNAs related to proliferation, invasion, and the ability to form metastases in various types of cancer are microRNAs (miRNAs) and circular RNAs (circRNAs). miRNAs are small sequence of approximately 22 nucleotides that negatively control gene expression at the post-transcriptional level by binding to the 3'-UTR region of their target mRNAs. CircRNAs, on the other hand, are highly resistant single-stranded circular molecules that play an important role in the regulation of mRNA levels due to their function as miRNA sponges. This function makes them potential candidates for therapeutic applications, particularly in situations where innate or acquired resistance to all existing treatment modalities poses challenges for certain cancers, including melanoma. To date, several circRNAs that contribute to its development have been identified, including hsa_circ_0079593. This circRNA is expressed at higher levels in melanoma than in healthy tissues and is associated with increased metastasis and malignant melanoma progression through the regulation of multiple "sponging" axes. Examples of these interactions are the circ_0079593/miR-573/ABHD2 and circ_0079593/miR-516b/GRM3 axes. In this study, we investigated whether circ_0079593 is involved in the development and aggressiveness of melanoma by regulating CHAF1B and MCAM genes through the inhibition of miR-516b-5p. To determine this, we first investigate the expression of circRNA-0079593 and miR-516b-5p in a healthy melanocyte cell line, and in some melanoma metastatic cell lines. Subsequently, I used in vitro assays to examine their regulatory effect on CHAF1B and MCAM genes, both of which showed increased expression linked to a more aggressive and invasive melanoma phenotype. This investigation gives significant insights but needs further investigation to assess the diagnostic and therapeutic implications of this network of interactions in the context of melanoma.
L’attribuzione dell’importante ruolo regolatorio dei RNA non codificanti (ncRNA) ed il loro coinvolgimento nella progressione tumorale è avvenuta solo negli ultimi decenni. Ad oggi, tra le classi di ncRNA maggiormente studiate in relazione alla proliferazione, all’invasione e alla capacità di formare metastasi in vari tipi di cancro troviamo i microRNA (miRNA) e i circRNA. I miRNA sono piccole sequenze di circa 22 nucleotidi che controllano negativamente l'espressione genica a livello post-trascrizionale legandosi alla regione 3'-UTR dei loro mRNA bersaglio. I circRNA, invece, sono molecole circolari a singolo filamento molto resistenti che svolgono un ruolo importante nella regolazione dei livelli di mRNA grazie alla loro funzione di spugne di miRNA. Questa funzione li rende potenziali candidati per applicazioni terapeutiche, in particolare in situazioni in cui la resistenza innata o acquisita a tutte le modalità di trattamento esistenti pone sfide in alcuni tipi di cancro, incluso il melanoma. Ad oggi, sono stati identificati diversi circRNA che contribuiscono allo sviluppo del melanoma, tra cui hsa_circ_0079593. Quest'ultimo risulta espresso ad alti livelli nel melanoma rispetto ai tessuti sani ed è associato ad un aumento delle metastasi e della progressione del melanoma maligno attraverso la regolazione di molteplici assi di "sponging": circ_0079593/miR-573/ABHD2 e circ_0079593/miR-516b/GRM3 sono due esempi. In questo studio, il mio obbiettivo è stato verificare se il circ_0079593 è coinvolto nella progressione allo sviluppo e all'aggressività del melanoma attraverso la regolazione dei geni CHAF1B e MCAM tramite l’inibizione del miR-516b-5p. Per farlo, ho innanzitutto verificato l'espressione del circ_0079593 e del miR-516b-5p in una linea cellulare di melanociti sani e in alcune linee di metastasi di melanoma. Successivamente, ho utilizzato saggi i vitro per esaminare il loro effetto regolatorio sui geni di CHAF1B e MCAM, la cui elevata espressione è associata ad un fenotipo aggressivo e invasivo del melanoma. Questa indagine fornisce spunti significativi, ma necessita di ulteriori approfondimenti per valutare le prospettive diagnostiche e terapeutiche derivanti da questa rete di interazioni nel contesto del melanoma.
Circular RNA circ_0079593 enhances malignant melanoma progression by the regulation of miR-516b-5p/MCAM and miR-516b-5p/CHAF1B axes.
De Tomi Elisa
2023-01-01
Abstract
The regulatory role of non-coding RNAs (ncRNAs) and their implication in tumor progression have only occurred in recent decades. To date, among the most studied classes of ncRNAs related to proliferation, invasion, and the ability to form metastases in various types of cancer are microRNAs (miRNAs) and circular RNAs (circRNAs). miRNAs are small sequence of approximately 22 nucleotides that negatively control gene expression at the post-transcriptional level by binding to the 3'-UTR region of their target mRNAs. CircRNAs, on the other hand, are highly resistant single-stranded circular molecules that play an important role in the regulation of mRNA levels due to their function as miRNA sponges. This function makes them potential candidates for therapeutic applications, particularly in situations where innate or acquired resistance to all existing treatment modalities poses challenges for certain cancers, including melanoma. To date, several circRNAs that contribute to its development have been identified, including hsa_circ_0079593. This circRNA is expressed at higher levels in melanoma than in healthy tissues and is associated with increased metastasis and malignant melanoma progression through the regulation of multiple "sponging" axes. Examples of these interactions are the circ_0079593/miR-573/ABHD2 and circ_0079593/miR-516b/GRM3 axes. In this study, we investigated whether circ_0079593 is involved in the development and aggressiveness of melanoma by regulating CHAF1B and MCAM genes through the inhibition of miR-516b-5p. To determine this, we first investigate the expression of circRNA-0079593 and miR-516b-5p in a healthy melanocyte cell line, and in some melanoma metastatic cell lines. Subsequently, I used in vitro assays to examine their regulatory effect on CHAF1B and MCAM genes, both of which showed increased expression linked to a more aggressive and invasive melanoma phenotype. This investigation gives significant insights but needs further investigation to assess the diagnostic and therapeutic implications of this network of interactions in the context of melanoma.File | Dimensione | Formato | |
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PhD_Thesis_Elisa_De_Tomi_Univr.pdf
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