Cristian Andrade-Madrigal

Acetylcholine (ACh) and prolactin (PRL) are increasingly recognized as additional factors potentially influencing breast cancer biology. Their effects may depend on tumor-cell context and on the molecular signaling state of individual breast cancer subtypes. This study aimed to evaluate phosphorylation responses to ACh and PRL and examine transcriptomic associations involving related signaling genes. MCF7, T47D, and MDA-MB-231 breast cancer cell lines were treated with ACh, PRL, or combined ACh + PRL for 4 h. Significant changes in selected phosphoproteins were summarized for proteins related to AKT–S6 kinase, EGFR–ERK, TP53–Src/PLC-γ1, and STAT3–JUN signaling. Transcriptomic analyses evaluated associations between acetylcholinesterase (ACHE) or prolactin receptor (PRLR) expression and selected signaling-related genes in breast cancer cohorts, including subtype-specific analyses, tumor-versus-normal comparisons, and overall-survival associations. ACh and PRL produced distinct, cell-line-specific phosphorylation patterns. The MCF7 cell line showed increased AKT, p70S6K, and p53 phosphorylation after ACh, whereas combined treatment increased AKT and STAT3 phosphorylation. The T47D cell line showed a mixed response, with combined treatment increasing p70S6K, PLC-γ1, and Src phosphorylation but reducing AKT, ERK1/2, and c-Jun phosphorylation. The MDA-MB-231 cell line showed the broadest response, with combined ACh + PRL increasing AKT, RSK, STAT3, and c-Jun phosphorylation. Transcriptomic analyses identified subtype-dependent associations of ACHE and PRLR with genes encoding AKT, S6 kinase, ERK, Src/PLC-γ1, STAT3, and JUN-related proteins. Several genes also showed differential tumor-versus-normal expression or survival associations. ACh and PRL are associated with context-dependent signaling responses in breast cancer. Experimental phosphoprotein changes and tumor transcriptomic associations converged on related signaling modules but represented different molecular levels and did not establish complete pathway activation or direct causality. These findings support further investigation of ACh–PRL interactions and their potential contribution to breast cancer biology.

Chronic exposure to benzo[a]pyrene (BaP), a Group 1 IARC carcinogen, is a major driver of lung carcinogenesis; however, how sustained subcytotoxic exposure reprograms bronchial epithelium toward preneoplastic states remains poorly defined. Here, we subjected BEAS-2B human bronchial epithelial cells to 12 weeks of continuous BaP at environmentally relevant concentrations (0.1 and 1.0 µM) and interrogated the resulting phenotypes using an integrated multi-scale framework encompassing functional toxicology, RT-qPCR, RNA-seq, phospho-kinase/NF-κB arrays, and organotypic air–liquid interface (ALI) cultures. Cells maintained metabolic competence throughout, evidenced by sustained CYP1A1 and CYP1B1 induction at both acute (4 h) and chronic (12-week) timepoints, while accumulating genotoxic stress as demonstrated by dose-dependent nuclear γ-H2AX foci formation and ATM phosphorylation (Ser1981). RNA-seq revealed a dose-dependent transcriptional shift: 0.1 µM BaP yielded 119 differentially expressed genes (DEGs; |log2FC| ≥ 1, FDR < 0.05), whereas 1.0 µM generated 255 DEGs. Downregulated transcripts were enriched for extracellular matrix and cell-adhesion programs (COL14A1, ADAMTS2, CSMD3, CADM3), while upregulated genes encompassed inflammatory, calcium-signaling, and vesicle-trafficking modules (NFATC4, CSF2RA, SYT1, PCLO). Phospho-kinase/NF-κB arrays confirmed a p53/NF-κB signaling nexus, with concurrent activation of MAPK/ERK (Thr202/Tyr204) and PI3K/Akt (Ser473) pathways. Despite persistent genotoxic stress, cells did not acquire anchorage-independent growth and remained non-tumorigenic in vivo. Critically, ALI organotypic cultures derived from BaP-exposed cells exhibited histological dysplasia, nuclear pleomorphism, and disrupted apical-basal polarity. These findings mechanistically link chronic BaP exposure to an initiation-like preneoplastic state and establish a validated 2D/3D multi-omics platform for PAH-driven lung carcinogenesis research.

Torque teno virus (TTV) is a highly prevalent DNA virus in humans, but its role in carcinogenesis is not well understood. While human papillomavirus (HPV) is a well-established etiological agent in cervical cancer, co-infections with other viruses such as Epstein–Barr virus (EBV) or TTV may influence disease progression. We conducted a cross-sectional study using 94 formalin-fixed, paraffin-embedded (FFPE) cervical tissue samples. These specimens were collected from women with cervical intraepithelial lesions (CINI-III) or squamous cell carcinoma (SCC) at the Clinical Hospital of the University of Chile. After extracting DNA, we screened for TTV using real-time polymerase chain reaction (qPCR). Statistical analysis was performed using Fisher’s exact test. Of the 94 samples, 83 were positive for the human β-globin gene and included in the final analysis. TTV was detected in 12.0% (10/83) of these samples. Among the TTV-positive cases, the virus was most frequently detected in high-grade lesions (70.0%), followed by low-grade lesions (20.0%) and squamous cell carcinoma (10.0%). However, these differences were not statistically significant (p = 0.688). This is the first study to assess TTV prevalence in cervical lesions among Chilean women. Although we found no statistically significant associations, a higher frequency of TTV was detected in precursor lesions compared to SCC. Further studies are needed to understand the potential immunomodulatory role of TTV in cervical carcinogenesis.

Xenobiotics are non-natural chemical compounds to which the human population is exposed. Chronic exposure to certain xenobiotics is associated with various diseases, including cancer development. Anelloviruses (AVs), including Torque Teno Virus (TTV), Torque Teno Mini Virus (TTMV), and Torque Teno Midi Virus (TTMDV), are ubiquitous viruses found in the general population. As no disease has been definitively associated with AVs, they are sometimes referred to as “viruses awaiting a disease”. This review explores the potential roles of xenobiotics and AVs in colorectal cancer (CRC) development and suggests a potential interplay between them. Evidence suggests an association between certain xenobiotics (like pesticides, cigarette smoke components, and dietary factors) and CRC, while such an association is less clear for AVs. The high prevalence of AVs suggests these infections alone may be insufficient to disrupt homeostasis; thus, additional factors might be required to promote disease, potentially including cancer.

High-risk human papillomaviruses (HR-HPVs) are the etiological agents of cervical, anogenital, and a subset of oropharyngeal cancers. In addition, HR-HPVs have been detected in lung carcinomas worldwide, even though the role of these viruses in this type of cancer is not fully understood. This study evaluated the presence of HPV in a cohort of 204 lung cancer cases by multiplex polymerase chain reaction (PCR)-Luminex. In addition, we used transcriptomic approaches to characterize the HPV-associated gene expression profile in the context of tobacco-smoke-associated lung cancer. HPV16 was detected in 8/204 lung carcinomas (4.0%). Through a significance analysis of microarrays (SAM) analysis, we found that the solute carrier family 7-member 11 (SLC7A11/xCT) gene (an antiporter that mediates the uptake of extracellular cystine) is up-regulated in tobacco-smoke- and HPV-associated lung cancers. In addition, SLC7A11 up-regulation correlates with both HR-HPV16 E6/E7 expression and tobacco smoke exposure in lung epithelial cells. Furthermore, we found decreased survival in HPV/SLC7A11-positive patients with lung cancer when compared to HPV/SLC7A11-negative cases. Thus, this study suggests that SLC7A11 up-regulation is associated with both HPV-positive and tobacco-smoke-associated lung carcinomas, with a potential association with clinical prognosis.