SPOCK2 gene expression is downregulated in pancreatic ductal adenocarcinoma cells and correlates with prognosis of patients with pancreatic cancer

• Published in: Journal of cancer research and clinical oncology 2023-09, Vol.149 (11), p.9191-9200
• Main Authors: Aghamaliyev, Ughur, Su, Kaifeng, Weniger, Maximilian, Koch, Dominik, D‘Haese, Jan G., Werner, Jens, Bazhin, Alexandr V.
• Format: Article
• Language: English
• Subjects: 5-aza-2'-deoxycytidine; Adenocarcinoma; Azacytidine; Cancer Research; Carcinoma, Pancreatic Ductal - pathology; Cell Line, Tumor; Cell migration; Cell Movement - genetics; Cell Proliferation; Demethylation; Down-regulation; Drug resistance; Extracellular matrix; Gene Expression; Gene Expression Regulation, Neoplastic; Hematology; Humans; Internal Medicine; Medical prognosis; Medicine; Medicine & Public Health; Oncology; Pancreatic cancer; Pancreatic Neoplasms - pathology; Prognosis; Proteoglycans - genetics; Proteoglycans - metabolism; Proteoglycans - therapeutic use; RNA, Small Interfering - genetics; RNA, Small Interfering - therapeutic use; siRNA; Transfection; Tumorigenicity
• ISSN: 0171-5216; 1432-1335; 1432-1335
• DOI: 10.1007/s00432-023-04845-5

Objectives Pancreatic ductal adenocarcinoma (PDAC) represents a widespread form of malignant pancreatic neoplasms and a leading oncologic cause of death in Europe and the USA. Despite advances in understanding its molecular biology, the 5-year survival rate remains low at 10%. The extracellular matrix in PDAC contains proteins, including SPOCK2, which are essential for tumorigenicity and drug resistance. The present study aims to explore the possible role of SPOCK2 in the pathogenesis of PDAC. Materials and methods Expression of SPOCK2 was evaluated in 7 PDAC cell lines and 1 normal pancreatic cell line using quantitative RT-PCR. Demethylation of the gene was carried out using 5-aza-2'-deoxycytidine (5-aza-dC) treatment with subsequent validation Western Blot analysis. In vitro downregulation of SPOCK2 gene was performed using siRNA transfection. MTT and transwell assays were employed to evaluate the impact of the SPOK2 demethylation on the proliferation and migration of PDAC cells. KM Plotter was applied to analyze a correlation between SPOCK2 mRNA expression and the survival of PDAC patients. Results In contrast to the normal pancreatic cell line, SPOCK2 expression was significantly downregulated in PDAC cell lines. Treatment with 5-aza-dC, led to increase in SPOCK2 expression in the cell lines tested. Importantly, compared with control cells, transfected with SPOCK2 siRNA cells exhibited increased growth rates and more migration ability. Finally, we demonstrated that a high SPOCK2 expression level correlated with longer overall survival of patients with PDAC. Conclusion The expression of SPOCK2 is downregulated in PDAC as a result of hypermethylation of its corresponding gene. SPOCK2 expression as well as the demethylation of its gene could be a potential marker for PDAC.