English abstract
Ovarian cancer (OC) poses a significant challenge to women's health globally, contributing to high rates of cancer-related morbidity and mortality. Most OC cases are diagnosed at advanced stages, resulting in low survival rates. Therefore, it is crucial to identify effective screening methods for early detection. The discovery of long non coding RNAs (lncRNAs) as key regulators of gene expression has opened up new avenues for investigating the molecular complexities of cancer biology. Bioinformatic analysis revealed higher expression of the lncRNA SOCS3-DT in OC recurrent samples, suggesting its potential as a biomarker. However, the precise functional role of SOCS3 DT in OC remains unclear. In this study, we used qRT-PCR to monitor the expression levels of SOCS3-DT in a panel of OC cell lines and OC tissue samples from our tissue bank at SQUH, and from Origene company. Furthermore, we silenced SOCS3-DT expression in OVSAHO cells and subsequently evaluated its impact on proliferation, colony formation, and wound healing assays. Additionally, to assess the effect of SOCS3 DT knockdown on key markers associated with cancer pathways, we performed qRT PCR and western blotting analysis. In cell lines, we observed overexpression of SOCS3 DT in the high-grade OC cell line, OVSAHO (p= 0.034). Conversely, the low-grade OC cell lines (OSE 1, MCAS, A2780 CP, A2780 S, and SKOV3) exhibited downregulated expression of SOCS3-DT compared to the normal ovarian cell line (HOSE 6-3) at the RNA level. Based on these findings, the OVSAHO cell line was selected for further functional analysis. Expression analysis using qRT-PCR revealed higher levels of SOCS3-DT in the early and late stages of OC patients compared to healthy individuals, V indicating its potential as a promising marker for OC detection. Upon silencing SOCS3 DT in OVSAHO cells, we observed reduced cell proliferation and decreased colony formation ability compared to the negative control. However, no significant difference was observed in the wound-healing ability between the silenced group and the control group. Moreover, regarding the expression of the pro-apoptotic marker, knocking down SOCS3-DT reduced the expression of CASPASE-3 at RNA and protein levels and CLEAVE CASPASE-9 at the protein level. However, the anti-apoptotic marker BCL2 expression was only downregulated at the RNA level. The expression of the autophagy markers (ATG5 and BECLIN) was reduced in siSOCS3-DT treated OVSAHO cells compared to NC at the RNA level, while the level of BECLIN only was lower at the protein level. In terms of EMT markers, the expression of the epithelial marker (CDH1) was reduced at the RNA level, while the mesenchymal markers (VIMENTIN and CDH2) were downregulated at the protein level. However, there was no significant change in the expression of CASPASE 9, BAX, CYCLIN D1, and CDH2 at the RNA levels and CDK4, CDK6, ATG5, BCL2, and CYCLIN D1 at the protein level. In conclusion, this study offers novel insight into discovering a new effective detection biomarker (SOCS3-DT) involved in the early stages of the disease, with the potential to provide a novel therapeutic target and approaches for the efficient treatment of this devastating disease. Moreover, the differential expression levels of the key cancer-associated genes highlight the involvement of SOCS3-DT in apoptosis and autophagy pathways. In addition, the variation in the expression of EMT markers suggests the role of SOCS3-DT in cancer metastasis.
