Julio César Villegas-Pineda

Phenotypic Heterogeneity of Cancer Associated Fibroblasts in Cervical Cancer Progression: FAP as a Central Activation Marker

Cervical cancer (CC) is the fourth leading cancer among women and is one of the principal gynecological malignancies. In the tumor microenvironment, cancer-associated fibroblasts (CAFs) play a crucial role during malignant progression, exhibiting a variety of heterogeneous phenotypes. CAFs express phenotypic markers like fibroblast activation protein (FAP), vimentin, S100A4, α-smooth muscle actin (αSMA), and functional markers such as MMP9. This study aimed to evaluate the protein expression of vimentin, S100A4, αSMA, FAP, and MMP9 in mesenchymal stem cells (MSC)-CAF cells, as well as in cervical cancer samples. MSC cells were stimulated with HeLa and SiHa tumor cell supernatants, followed by protein evaluation and cytokine profile to confirm differentiation towards a CAF phenotype. In addition, automated immunohistochemistry (IHQa) was performed to evaluate the expression of these proteins in CC samples at different stages. Our findings revealed a high expression of FAP in stimulated MSC cells, accompanied by the secretion of pro/anti-inflammatory cytokines. In the other hand, CC samples were observed to have high expression of FAP, vimentin, αSMA, and MMP9. Most importantly, there was a high expression of their activation proteins αSMA and FAP during the different stages. In the early stages, a myofibroblast-like phenotype (CAFs αSMA+ FAP+), and in the late stages a protumoral phenotype (CAF αSMA− FAP+). In summary, FAP has a crucial role in the activation of CAFs during cervical cancer progression.

Cervical Cancer Biomarkers in Non-Cervical Samples: Emerging Tools for Diagnosis and Prognosis

Cervical cancer (CC) is the gynecological cancer with the highest incidence and mortality worldwide. High-risk oncogenic human papillomaviruses (HPV) genotypes 16 and 18 are the primary risk factors for developing this female neoplasm, with them being the etiological agents of 70% of cervical cancers. Despite the availability of various prevention strategies, laboratory tests capable of detecting the disease in its previous and early stages, and multiple treatment schemes, CC incidence and mortality rates remain high, due in part to the population’s rejection or disinterest in the current type of sampling. An alternative that could encourage women to take better care of their gynecological health is the availability of tests that detect biomarkers in non-cervical samples with high sensitivity and specificity. The detection of biomarkers in non-cervical samples (blood, serum, plasma, urine, and vaginal fluids) may help reduce the discomfort associated with cervical sampling in patients, therefore promoting gynecological healthcare. This review discusses current diagnostic methods and recent advances in CC biomarkers detected in non-cervical samples, emphasizing their potential for diagnosis, prognosis, and patient monitoring. We further discuss the challenges and future perspectives of applying these biomarkers in clinical practice. The results of this review show that there is a considerable range of biomarkers proposed as alternative tools with high efficacy. Their identification in previous stages of the disease and routinely in non-cervical samples could help reduce the incidence and mortality rates of CC.

Overexpression of Fut 2, 4, and 8, and nuclear localization of Fut 4 in ovarian cancer cell lines induced by ascitic fluids from epithelial ovarian cancer patients

AbstractFucosyltransferases (Fut) regulate the fucosylation process associated with tumorogenesis in different cancer types. Ascitic fluid (AF) from patients diagnosed with advanced stage of epithelial ovarian cancer (EOC) is considered as a dynamic tumor microenvironment associated with poor prognosis. Previous studies from our laboratory showed increased fucosylation in SKOV‐3 and OVCAR‐3, cancer‐derived cell lines, when these cells were incubated with AFs derived from patients diagnosed with EOC. In the present work we studied three fucosyltransferases (Fut 2, Fut 4, and Fut 8) in SKOV‐3, OVCAR‐3 and CAOV‐3 cell lines in combination with five different AFs from patients diagnosed with this disease, confirming that all tested AFs increased fucosylation. Then, we demonstrate that mRNAs of these three enzymes were overexpressed in the three cell lines under treatment with AFs. SKOV‐3 showed the higher overexpression of Fut 2, Fut 4, and Fut 8 in comparison with the control condition. We further confirmed, in the SKOV‐3 cell line, by endpoint PCR, WB, and confocal microscopy, that the three enzymes were overexpressed, being Fut 4 the most overexpressed enzyme compared to Fut 2 and Fut 8. These enzymes were concentrated in vesicular structures with a homogeneous distribution pattern throughout the cytoplasm. Moreover, we found that among the three enzymes, only Fut 4 was located inside the nuclei. The nuclear location of Fut 4 was confirmed for the three cell lines. These results allow to propose Fut 2, Fut 4, and Fut 8 as potential targets for EOC treatment or as diagnostic tools for this disease.