Gene Expression of TGF-Β and Nf-Κb in Penile Cancer Samples Infected With HPV in Maranhão

Penile cancer (PC) is a rare neoplasm worldwide but has a high incidence in regions with low socioeconomic development. HPV is a DNA virus with more than 200 strains, with subtypes 16 and 18 being the main oncogenic types. In this context, the study aimed to evaluate the gene expression of two inflammation-modulating pathways in HPV-positive penile cancer samples. This was a cross-sectional study with collection of tumor tissue and adjacent non-tumor tissue samples from patients who underwent penectomy (n = 11). DNA was extracted using a specific kit and samples were evaluated for viral presence using the Seegene Anyplex II HPV28 Detection multiplex qPCR technique. Negative samples were further analyzed for viral presence using nested PCR. RNA was extracted using a specific kit, converted to cDNA, and gene expression of TGF-β and NF-κB was analyzed by qPCR. Results were analyzed using the 2^-ΔΔCT method. Statistical analysis was performed using GraphPad Prism (v10). By multiplex testing, 2 patients were HPV-positive in both tumor and adjacent tissue; 2 were positive only in adjacent tissue; and 7 had both samples negative. Among positive samples, HPV subtypes 6, 16, 31, 40, 42, 43, and 73 were identified. One patient with subtype 16 also presented coinfection with low-risk subtypes. Negative samples were analyzed by nested PCR. The 2 patients with positive adjacent tissue also tested positive in tumor tissue. Of the 7 initially negative cases, 5 tested positive for HPV (2 in tumor tissue only and 3 in adjacent tissue only), and 2 samples were excluded due to insufficient material. Regarding gene expression, patients with HPV-positive tumor and adjacent tissue samples (n = 4) showed higher NF-κB expression and lower TGF-β expression. In contrast, patients HPV-positive only in adjacent tissue (n = 3) showed higher TGF-β expression than NF-κB. Patients HPV-positive only in tumor tissue (n = 2) showed similar average expression of both genes. The data highlight differences in sensitivity between HPV detection methods: multiplex PCR allows HPV subtype genotyping, while nested PCR shows high sensitivity without subtype identification. Regarding gene expression, the presence of the virus in tumor tissue appears to induce an NF-κB – dependent inflammatory pattern, modulating the expression of inflammation-related genes and influencing the tumor microenvironment.