In silico karakterizacija miRNK meta potpisa i validacija odabranih miRNK kao potencijalnih prognostičkih biomarkera u oralnom karcinomu

Abstract

Oko 30% pacijenata sa oralnim karcinomom će nakon ekscizije tumora imati recidiv u periodu od dve godine. Stoga je važno identifikovati senzitivne molekularne biomarkere koji mogu imati dijagnostički i prognostički značaj. Proces oralne karcinogeneze je povezan sa deregulacijom ekspresije malih nekodirajućih miRNK molekula. Cilj istraživanja je bilo utvrđivanje najčešće deregulisanih miRNK u oralnom karcinomu i ispitivanje dijagnostičkog i prognostičkog značaja odabranih miRNK. Lista najčešće deregulisanih miRNK u oralnom karcinomu, miRNK meta potpis, je dobijena sprovođenjem meta-analize ranije publikovanih studija. Meta potpis najčešće deregulisanih miRNK u oralnom karcinomu čini 13 miRNK, od toga sedam povišeno (miR-21-5p, miR-31-3p, miR-135b-5p, miR-31-5p, miR-424-5p, miR-18a-5p, miR-21-3p) i pet sniženo eksprimiranih (miR-139-5p, miR-30a-3p, miR-375-3p, miR-376c-3p, miR-30a-5p). Studijsku grupu je činilo 35 pacijenata sa oralnim karcinomom. Na kliničkim uzorcima je validirana povišena ekspresija miR-31-3p, miR-135b-5p i snižena ekspresija miR-139-5p i miR-30a-5p gena u tkivu oralnog karcinoma. Panel od tri analizirane miRNK, miR-31-3p, miR-139-5p, miR-30a-5p su imale najbolji dijagnostički potencijal za diskriminaciju tkiva oralnog karcinoma od netumorskog tkiva (AUC: 0,780 (95% CI: 0,673-0,886), p<0,0005, senzitivnost 94,3%, specifičnost 51,4%). Značajno lošije preživljavanje su imali pacijenti sa povišenom ekspresijom miR-135b-5p, miR-18a-5p i miR-30a-5p (p=0,003, p=0,048, p=0,016, redom). Panel koji čine miR-31-3p, miR-139-5p, miR-30a-5p se može koristiti kao potencijalni dijagnostički set biomarkera za senzitivnu diskriminaciju tkiva oralnog karcinoma od netumorskog tkiva. Kao potencijalni prognostički biomarkeri se mogu koristiti miR-135b-5p, miR-18a-5p i miR-30a-5p. Rezultati ove doktorske disertacije predstavljaju dobar osnov za planiranje budućih istraživanja, koja bi trebalo sprovesti na većoj studijskoj grupi sa dužim vremenom praćenja pacijenata.

About 30% of patients with oral cancer will have a recurrence after the excision of the tumor within a period of two years. Therefore, it is important to identify sensitive molecular biomarkers that may have diagnostic and prognostic significance. The process of oral carcinogenesis is associated with the deregulation of the expression of small non-coding miRNA molecules. The aim of the study was to determine the most commonly deregulated miRNAs in oral cancer and to examine the diagnostic and prognostic significance of selected miRNAs. A list of the most commonly deregulated miRNAs in oral cancer, the miRNA meta signature, was obtained by conducting a meta-analysis of previously published studies. The meta signature of the most commonly deregulated miRNAs in oral cancer consisted of 13 miRNAs, of which seven were up- (miR-21-5p, miR-31-3p, miR-135b-5p, miR-31-5p, miR-424-5p, miR-18a-5p, miR-21-3p) and five down-regulated (miR-139-5p, miR-30a-3p, miR-375-3p, miR-376c-3p, miR-30a-5p). The study group consisted of 35 patients with oral cancer. Increased expression of miR-31-3p, miR-135b-5p and decreased expression of miR-139-5p and miR-30a-5p genes in oral cancer tissue were validated on clinical samples. A panel of three analyzed miRNAs, miR-31-3p, miR-139-5p, miR-30a-5p had the best diagnostic potential for discriminating oral cancer tissue from non-tumor tissue (AUC: 0.780 (95% CI: 0.673-0.886), p <0.0005, sensitivity 94.3%, specificity 51.4%). Patients with increased expression of miR-135b-5p, miR-18a-5p and miR-30a-5p had significantly worse survival (p=0.003, p=0.048, p=0.016, respectively). The panel consisting of miR-31-3p, miR-139-5p, miR-30a-5p can be used as a potential diagnostic biomarker set for sensitive discrimination of oral cancer tissue from non-tumor tissue. miR-135b-5p, miR-18a-5p and miR-30a-5p can be used as potential prognostic biomarkers. The results of this doctoral dissertation represent a good basis for planning future research, which should be conducted in a larger study group with a longer patient follow-up time.