Head and neck squamous cell carcinoma (HNSCC) is a group of cancers originating from the mucosal epithelium in the oral cavity, larynx, oropharynx, nasopharynx, and hypopharynx. Molecular factors can be key in the diagnosis, prognosis, and treatment of HNSCC patients. Long non-coding RNAs (lncRNAs) are molecular regulators composed of 200 to 100,000 nucleotides that act on the modulation of genes that activate signaling pathways associated with oncogenic processes such as proliferation, migration, invasion, and metastasis in tumor cells. However, up until now, few studies have discussed the participation of lncRNAs in modeling the tumor microenvironment (TME) to generate a protumor or antitumor environment. Nevertheless, some immune-related lncRNAs have clinical relevance, since AL139158.2, AL031985.3, AC104794.2, AC099343.3, AL357519.1, SBDSP1, AS1AC108010.1, and TM4SF19-AS1 have been associated with overall survival (OS). MANCR is also related to poor OS and disease-specific survival. MiR31HG, TM4SF19-AS1, and LINC01123 are associated with poor prognosis. Meanwhile, LINC02195 and TRG-AS1 overexpression is associated with favorable prognosis. Moreover, ANRIL lncRNA induces resistance to cisplatin by inhibiting apoptosis. A superior understanding of the molecular mechanisms of lncRNAs that modify the characteristics of TME could contribute to increasing the efficacy of immunotherapy.

Background: E2 is fundamental to the onset and progression of CC. CAFs are the most abundant stromal population in the tumor microenvironment. CAFs have been shown to promote protumoral effects in CC cell lines; however, the effects of E2-stimulated CAFs remain to be explored. Objectives: This study aimed to evaluate whether E2-stimulated CAFs modify the behavior of SiHa and HeLa tumor cells, including their metabolism, ROS production, migration, apoptosis, and gene expression. Methods: Characterization and estrogen receptor expression in primary CAF cultures were evaluated by immunofluorescence. Supernatants from non-stimulated and E2-stimulated CAFs were used to culture CC cells, and mitochondrial metabolism was measured by MTT, ROS production by H2DCFDA, migration using the wound-healing assay, apoptosis by Annexin V assay, and gene expression by next-generation RNA-seq. Results: Stimulation of CC cells with CAFs’ supernatants demonstrated that it influences tumor cells in different ways. In SiHa cells, the metabolic shift was supported by decreased mitochondrial metabolism, increased ROS production, and enhanced gene enrichment for glycolysis and hypoxia responses. On the other hand, in HeLa cells, CAFs enhanced migration and gene enrichment in KRAS signaling, epithelial-mesenchymal transition, and the proinflammatory response, via enrichment of the IL6/JAK/STAT3 signaling pathway, along with the overexpression of cytokines such as IFN-γ, IL-6, and IL-8. Conclusions: CAFs exert protumoral effects by promoting a metabolic shift in SiHa cells or enhancing migration and cytokine secretion in HeLa cells; these effects are independent of E2 stimulation in CAFs.

Background Cervical cancer (CC) remains one of the leading causes of morbidity and mortality in middle- and low-income countries. The tumor microenvironment (TME), especially cancer-associated fibroblasts (CAF), plays a key role in tumor development and progression through diverse mechanisms, including cytokine secretion, angiogenesis, metabolism, and immune modulation. Pioglitazone (Pgz) has shown anti-fibrotic effects, preventing fibroblast activation and differentiation into myofibroblasts; anti-inflammatory effects, reducing the expression of IL-1, IL-6, and TNFα; and even anti-tumor properties, decreasing tumor cell migration and proliferation. In our study, we investigated the impact of Pgz on CAF-mediated effects on HeLa cervical cancer cells, focusing on metabolism, migration, invasion, and proliferation, and on how Pgz affects HeLa cells’ ability to activate and induce a CAF phenotype. Methods After CAF obtention from cervical cancer biopsies, conditioned medium was collected from CAF or HeLa cell cultures after 24 h in free-serum media. Cells were treated with the conditioned media with or without pioglitazone for 24 h. To evaluate cell activity, we performed an MTT assay, measured ROS production, and glucose present in the medium. We also evaluate cell proliferation by bioimpedance, migration by a wound-healing assay, and invasion using the Matrigel and transwell systems. The cytokines produced by CAFs were evaluated by multiplex ELISA, and Mesenchymal Stem Cell differentiation into a CAF phenotype was evaluated by immunofluorescence for the markers αSMA and FAP. Statistical analyses were performed using GraphPad Prism software, using One-way analysis of variance with Tukey post hoc tests or the Kruskal-Wallis test for non-normal distributions. Results Our findings reveal that Pgz reduces CAF-induced metabolic activity, ROS production, glucose uptake, migration, invasion, and proliferation in HeLa cells. Additionally, Pgz suppresses HeLa-induced expression of αSMA and FAP in mesenchymal stem cells, suggesting it may help prevent CAF differentiation. Conclusions CAFs appear to be a promising target for future cancer therapies; disrupting their tumor-supportive role through Pgz might help slow cancer progression and improve therapeutic outcomes.