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<i>Background and Objectives:</i> Penile cancer is a rare neoplasm in developed countries with an incidence of 0.8/100,000 per male inhabitant. Despite the development of personalized medicine and multimodal treatment, the outcome of penile cancer treatment is insufficient. Our study aimed to assess the levels of pro-inflammatory cytokines’ mRNA such as interleukin 1-A (encoded by <i>IL1A</i> gene, alias <i>IL-1A)</i>, interleukin 1-B (<i>IL1B</i>, <i>IL-1B</i>), interleukin 1 receptor antagonist (<i>IL1RN</i>, <i>IL-1RN</i>), interleukin 6 (<i>IL6</i>, <i>IL-6</i>), transforming growth factor β1 (<i>TGFB1</i>, <i>TGFβ-1</i>), and Interferon-gamma (<i>INFG</i>, <i>INF-γ</i>) in penile cancer tissue and associate them with tumor progression and patient survival. <i>Material and Methods:</i> Skin biopsies from patients suffering from penile cancer (n = 6) and unchanged foreskin from 13 healthy adult males undergoing circumcision due to a short frenulum were obtained. Pro-inflammatory cytokine mRNA levels were quantified through qPCR. <i>Results:</i> We observed higher expression of pro-inflammatory cytokine genes (IL-1A, IL-1B, IL-6, INF-γ, TGF-β) in penile cancer tissue. The average follow-up period was 48 months (range: 38–54 months), during which only one penile tumor progression was observed However, this was without association with the nature of tumor (patient refused radical treatment). <i>Conclusions:</i> This is the first study to show increased expression of cytokines such as IL-1A, IL-1B, IL-6, INF-γ, and TGF-β in penile cancer with positive correlation between TNM staging and INF-γ levels in tumor samples (rs = 0.672, <i>p</i> = 0.045), which may be associated with the immunosuppressive role of the tumor environment.