Journal Article

Novel chemotherapeutic agent FX-9 activates NF-κB signaling and induces G1 phase arrest by activating CDKN1A in a human prostate cancer cell line

Koczan, D., Junghanss, C., Murua Escobar, H., Beller, M., Nolte, I., Wu, X.-F., Schille, J. T., Weiner, F., Hewicker-Trautwein, M.

2021

Overview

Background The aminoisoquinoline FX-9 shows pro-apoptotic and antimitotic effects against lymphoblastic leukemia cells and prostate adenocarcinoma cells. In contrast, decreased cytotoxic effects against non-neoplastic blood cells, chondrocytes, and fibroblasts were observed. However, the actual FX-9 molecular mode of action is currently not fully understood. Methods In this study, microarray gene expression analysis comparing FX-9 exposed and unexposed prostate cancer cells (PC-3 representing castration-resistant prostate cancer), followed by pathway analysis and gene annotation to functional processes were performed. Immunocytochemistry staining was performed with selected targets. Results Expression analysis revealed 0.83% of 21,448 differential expressed genes (DEGs) after 6-h exposure of FX-9 and 0.68% DEGs after 12-h exposure thereof. Functional annotation showed that FX-9 primarily caused an activation of inflammatory response by non-canonical nuclear factor-kappa B (NF-κB) signaling. The 6-h samples showed activation of the cell cycle inhibitor CDKN1A which might be involved in the secondary response in 12-h samples. This secondary response predominantly consisted of cell cycle-related changes, with further activation of CDKN1A and inhibition of the transcription factor E2F1 , including downstream target genes, resulting in G1-phase arrest. Matching our previous observations on cellular level senescence signaling pathways were also found enriched. To verify these results immunocytochemical staining of p21 Waf1/Cip1 ( CDKN1A ), E2F1 ( E2F1 ), PAI-1 ( SERPNE1 ), and NFkB2/NFkB p 100 ( NFKB2 ) was performed. Increased expression of p21 Waf1/Cip1 and NFkB2/NFkB p 100 after 24-h exposure to FX-9 was shown. E2F1 and PAI-1 showed no increased expression. Conclusions FX-9 induced G1-phase arrest of PC-3 cells through activation of the cell cycle inhibitor CDKN1A , which was initiated by an inflammatory response of noncanonical NF-κB signaling.

• Publisher: BioMed Central,Springer Nature B.V,BMC
• Subject: Adenocarcinoma / Androgens / Antimitotic agent / Antineoplastic Agents - pharmacology / Antineoplastic Agents - therapeutic use / Apoptosis / Biomedical and Life Sciences / Biomedicine / Blood cells / Cancer Research / Castration / Cell activation / Cell culture / Cell cycle / Chemotherapy / Chondrocytes / Cip protein / Cyclin-dependent kinase inhibitor p21 / Cyclin-Dependent Kinase Inhibitor p21 - metabolism / Cyclin-dependent kinases / Cytotoxicity / DNA microarrays / E2F1 Transcription Factor - antagonists & inhibitors / Fibroblasts / G1 phase / G1 Phase Cell Cycle Checkpoints - drug effects / G1 Phase Cell Cycle Checkpoints - genetics / G1-phase arrest / Gene expression / Gene Expression - drug effects / Gene Expression Profiling - methods / Health Promotion and Disease Prevention / Humans / Hybridization / Immunocytochemistry / Inflammation / Isoquinolinamine FX-9 / Isoquinolines - pharmacology / Isoquinolines - therapeutic use / Kinases / Laboratories / Lymphatic leukemia / Male / Medicine/Public Health / Microarray analysis / Middle Aged / Morphology / NF-kappa B - metabolism / NF-κB protein / NF-κB signaling / Oncology / PC-3 Cells / Plasminogen Activator Inhibitor 1 - metabolism / Prostate cancer / Prostatic Neoplasms, Castration-Resistant - genetics / Prostatic Neoplasms, Castration-Resistant - metabolism / S Phase Cell Cycle Checkpoints / Senescence / Signal transduction / Surgical Oncology / Time Factors / Tissue Array Analysis