Theranekron modulates extracellular matrix and apoptotic signaling pathways in colorectal cancer cells

Colorectal cancer (CRC) remains a major cause of cancer-related mortality worldwide, largely due to therapeutic resistance and the complexity of the tumor microenvironment. The extracellular matrix (ECM) critically regulates tumor progression by balancing matrix metalloproteinases (MMPs) and their inhibitors (TIMPs). Theranekron^® D6, an extract of Tarantula cubensis with known anti-inflammatory and wound-healing effects, has shown anticancer potential. This study investigated its molecular impact on ECM remodeling and apoptosis in human CRC. Human colorectal adenocarcinoma (Caco-2) and non-tumoral embryonic kidney (HEK293) cells were treated with Theranekron^® D6 (1-1000 µL/mL). Cell viability was evaluated using the MTT assay, revealing a dose- and time-dependent cytotoxic response with an IC₅₀ of 400 µL/mL at 48 h. Gene expression of 62 target genes spanning ECM remodeling, apoptosis, and key signaling axes (PI3K/AKT, NF-κB, and SMAD/STAT) was analyzed by quantitative polymerase chain reaction (qPCR), and protein levels of TIMP1, MMP2, MMP9, MMP14 and MMP28 were quantified by enzyme-linked immunosorbent assay (ELISA). Pathway interactions were assessed via STRING-based enrichment and clustering analyses. Theranekron^® markedly upregulated pyruvate dehydrogenase kinase 1 (PDK1) by 23-fold, with concurrent upregulation of PTEN suggesting a negative feedback mechanism that may limit net pro-survival PI3K/AKT output. This regulation restored a TIMP1-dominant ECM profile and reduced MMP2 and MMP28 protein levels. NF-κB pathway components were broadly downregulated at the transcriptional level. Concurrently, Theranekron^® induced transcriptional changes associated with mitochondrial and SMAD-dependent apoptotic pathways. Theranekron^® acts as a cell-context-specific modulator, simultaneously regulating PI3K/AKT/PDK1, NF-κB and SMAD pathways to restore ECM integrity and modulate apoptotic signaling pathways in Caco-2 CRC cells, suggesting its potential as a multi-target experimental therapeutic candidate in in vitro CRC models.