Olga Stasikowska-Kanicka

Background/Objectives: Actinic keratosis (AK) is a common ultraviolet-induced intraepidermal neoplasm with a recognized potential to progress to cutaneous squamous cell carcinoma (cSCC). Although photodynamic therapy (PDT) is an established field-directed treatment for AK, its effects on the inflammatory and angiogenic tumor microenvironment, particularly regarding mast cell (MC) infiltration and microvascular remodeling, remain incompletely understood. This study aimed to investigate the impact of PDT on mast cell (MC) count and CD31-associated microvessel density (MVD) in AK lesions. Methods: In this prospective single-center study, 31 patients with histopathologically confirmed AK underwent three sessions of 5-aminolevulinic acid-based PDT. Each patient contributed samples of clinically uninvolved skin, the target AK lesion before PDT, and the same AK lesion after PDT. Repeated tissue measurements were analyzed using the Friedman test followed by exact Wilcoxon signed-rank tests with Holm correction. Associations between MC count and MVD were assessed using Spearman rank correlations. Clinical and dermoscopic grades were analyzed as paired ordinal outcomes. Results: Significant clinical and dermoscopic improvement was observed in the majority of patients following PDT, with complete lesion clearance achieved in 17 of 31 patients (54.84%). Both MC count and MVD were significantly higher in AK lesions compared with clinically uninvolved skin (p < 0.05). PDT resulted in a significant reduction in both parameters (p < 0.05), although post-treatment levels remained elevated relative to clinically uninvolved skin. MC count and MVD were positively correlated both before and after PDT, with a stronger association observed after treatment. Conclusions: PDT demonstrates not only cytotoxic effects against dysplastic keratinocytes but also may significantly modulate the inflammatory and angiogenic microenvironment in AK. The persistent correlation between MC infiltration and neovascularization after therapy may reflect incomplete normalization of the local microenvironment despite clinical improvement. These findings suggest that MC count and MVD may merit further evaluation as adjunctive biomarkers of response in PDT-treated AK.

Background: Actinic keratoses (AKs) are common pre-neoplastic lesions that may progress to cutaneous squamous cell carcinoma (cSCC). Photodynamic therapy (PDT) is an effective field-directed treatment for AK, but its impact on key biomarkers remains unclear. This study evaluates the clinical, dermatoscopic, and immunohistochemical effects of PDT on AK, with a focus on proliferation (Ki67, p53) and inflammation (COX-2) markers, to assess its efficacy in delaying carcinogenesis. Methods: In our prospective one-center study, we enrolled 31 patients with AK, with no history of previous AK treatment. They underwent three PDT sessions at four-week intervals, with follow-up eight weeks after the final session. Clinical, dermatoscopic, and immunohistochemical analyses of Ki67, p53, and COX-2 expression were performed before and after treatment. Results: Clinically, 54.8% of patients achieved complete lesion clearance, with no residual severe AK lesions. Ki67 and p53 immunoexpression significantly decreased post-PDT (p < 0.05), confirming its antiproliferative effect. COX-2 expression also declined significantly (p < 0.05), supporting PDT’s anti-inflammatory role. However, COX-2 remained stable or increased in 35.48% of cases, possibly due to inflammation-induced regeneration. There is a positive correlation between the reduction in Ki67, p53, and COX-2 immunoexpression and the decrease in AK severity (both according to Olsen grade and dermatoscopic grade). Conclusions: PDT effectively reduces AK severity, proliferation, and inflammation markers, potentially delaying carcinogenesis. However, residual biomarker expression suggests that additional treatment sessions or combination therapies may be necessary for complete lesion clearance. Further studies are required to optimize PDT protocols.

During metastasis, cancer cells undergo phenotype changes in the epithelial-mesenchymal transition (EMT) process. Extracellular vesicles (EVs) released by cancer cells are the mediators of intercellular communication and play a role in metastatic process. Knowledge of factors that influence the modifications of the pre-metastatic niche for the migrating carcinoma cells is important for prevention of metastasis. We focus here on how cancer progression is affected by EVs released from either epithelial-like HT29-cells or from cells that are in early EMT stage triggered by Snail transcription factor (HT29-Snail). We found that EVs released from HT29-Snail, as compared to HT29-pcDNA cells, have a different microRNA profile. We observed the presence of interstitial pneumonias in the lungs of mice injected with HT29-Snail cells and the percent of mice with lung inflammation was higher after injection of HT29-Snail-EVs. Incorporation of EVs released from HT29-pcDNA, but not released from HT29-Snail, leads to the increased secretion of IL-8 from macrophages. We conclude that Snail modifications of CRC cells towards more invasive phenotype also alter the microRNA cargo of released EVs. The content of cell-released EVs may serve as a biomarker that denotes the stage of CRC and EVs-specific microRNAs may be a target to prevent cancer progression.

Background: Zinc (Zn) and selenium (Se) play a well-documented role in cancer prevention (e.g., for prostate cancer), and their combined supplementation is often given as a recommended prophylactic agent. The aim of the study was to determine the influence of Zn and/or Se supplementation on the androgen receptor (AR) in the prostate lobes and the serum selected hormone concentrations; a hitherto unresearched topic. Methods: Male rats (n = 84) were administered with Zn and/or Se intragastrically for up to 90 days. The effects of administration on the tested parameters were checked after 30 and 90 days of administration and additionally, 90 days after the end of 90 day administration. Results: Zn alone leads to an increase in serum testosterone concentrations, while the protein expression of AR in both parts of the prostate increases. Combined administration of Zn and Se eliminates the effect of Zn, which may suggest that these two elements act antagonistically. Se supplementation alone results in the same level of AR protein expression in administration and 90 days after administration periods. Conclusion: This paper presents the first report of the influence of Zn and/or Se supplementation on the protein expression of AR in the prostate. Our findings seem to indicate that simultaneous supplementation of both elements may be ineffective.

The study was aimed to evaluate the number of TAMs and to investigate whether they have association with microvessels density and patients’ survival times. 46 cases of melanomas, divided into four groups according to the Breslow scale, were tested immunohistochemically with antibodies...

Overexpression of inhibitory checkpoint PD1/PD‐L1 plays an important role in carcinogenesis and patients prognosis. 70 cases of oral squamous cell carcinoma (OSCC), 23 cases of oral leukoplakia (OLK), and 19 control cases were immunohistochemically stained with anti‐PD‐L1, ‐CD8, and ‐CD163 antibodies. PD‐L1 was expressed on dysplastic and subepithelial infiltrating cells of OLK as well as on cancer and tumor‐infiltrating cells of OSCC. In OSCC, PD‐L1 immunoexpression was significantly increased in comparison to OLK, and control groups. The correlative study showed significant correlations between the immunoexpression of PD‐L1 and the number of CD8+, CD163+ cells in both OLK and OSCC groups. We found also significant negative correlation between the number of PD‐L1+ infiltrating cells and the number of CD8+ cells in OSCC, and positive correlation between the number of PD‐L1+ infiltrating cells and CD163+ cells in OLK and OSCC groups. In conclusion, our study indicate that CD163+ and CD8+ infiltrating cells influence the early and subsequent stages of oral carcinogenesis. We demonstrated also that studied tumors may evade the host immune system by PD‐L1 immunoexpression not only on epithelial cells but on infiltrating cells as well.

Introduction. Neuropilins (NRPs) are multifunctional glycoproteins that play an important role in angiogenesis and cancer progression. The aim of the study was to examine the immunoexpression of neuropilin 1 (NRP1), the number of NRP1+ infiltrating cells and CD163+ macrophages, and density of microvessels (MVD) in oral squamous cell carcinoma (OSCC). Material and methods. The study was performed on 45 OSCC patients with metastases (OSCCM+), 51 patients without metastases (OSCCM-) and 17 control cases. The microvessels were identified by the presence of CD31 and the expression of the studied proteins was assessed by immunohistochemistry. Results. The immunoexpression of NRP1, the mean numbers of NRP1+, CD163+ infiltrating cells, and MVD were significantly increased in OSCCM+ patients in comparison to OSCCM-, and control groups. Moreover, in OSCCM- patients all these parameters were also significantly increased in comparison to controls. In OSCCM+ and OSCCM- groups, there were positive correlations between the immunoexpression of NRP1 and MVD (r = 0.41, p < 0.006; r = 0.51, p < 0.001, respectively), and between the number of NRP1+ infiltrating cells and CD163+ macrophages (r = 0.56, p < 0.001, r = 0.49, p < 0.001, respectively). Conclusions. The present study revealed overexpression of NRP1 in OSCC, especially in OSCC patients with metastasis, suggesting that NRP1 could potentially contribute to metastasis of oral cancer. The correlation between the number of NRP1+ infiltrating cells and CD163+ macrophages suggests that NRP1+ infiltrating macrophages are present in tumor microenvironment and may play a role in the progressions of oral cancer. Introduction. Neuropilins (NRPs) are multifunctional glycoproteins that play an important role in angiogenesis and cancer progression. The aim of the study was to examine the immunoexpression of neuropilin 1 (NRP1), the number of NRP1+ infiltrating cells and CD163+ macrophages, and density of microvessels (MVD) in oral squamous cell carcinoma (OSCC). Material and methods. The study was performed on 45 OSCC patients with metastases (OSCCM+), 51 patients without metastases (OSCCM-) and 17 control cases. The microvessels were identified by the presence of CD31 and the expression of the studied proteins was assessed by immunohistochemistry. Results. The immunoexpression of NRP1, the mean numbers of NRP1+, CD163+ infiltrating cells, and MVD were significantly increased in OSCCM+ patients in comparison to OSCCM-, and control groups. Moreover, in OSCCM- patients all these parameters were also significantly increased in comparison to controls. In OSCCM+ and OSCCM- groups, there were positive correlations between the immunoexpression of NRP1 and MVD (r = 0.41, p < 0.006; r = 0.51, p < 0.001, respectively), and between the number of NRP1+ infiltrating cells and CD163+ macrophages (r = 0.56, p < 0.001, r = 0.49, p < 0.001, respectively). Conclusions. The present study revealed overexpression of NRP1 in OSCC, especially in OSCC patients with metastasis, suggesting that NRP1 could potentially contribute to metastasis of oral cancer. The correlation between the number of NRP1+ infiltrating cells and CD163+ macrophages suggests that NRP1+ infiltrating macrophages are present in tumor microenvironment and may play a role in the progressions of oral cancer.

Background: The prognosis of human malignancies has been shown to depend on immunological parameters, such as macrophage polarization (M1 and M2). In this study, we identify the phenotype of macrophages, and investigate an involvement of infiltrated T cells that participate in the polarization of macrophages, in oral leukoplakia (OLK), and oral squamous cell carcinoma (OSCC).Methods: Immunohistochemical method was used to examine the number of CD68⁺, CD163⁺ (M2), iNOS⁺ (M1) macrophages, and CD4⁺, CD8⁺, CCR4⁺ (Th2), CCR5⁺ (Th1) cells in 102 cases of OSCC: without metastases—OSCC M(−) (n = 54), and with metastases—OSCC M(+) (n = 48), 23 cases of OLK, and 18 control cases.Results: The mean number of CD68⁺, CD163⁺, iNOS⁺, CD4⁺, CCR4⁺, CCR5⁺ cells was significantly increased in OSCC M(+) group compared with OLK, OSCC M(−) and control group. We found positive correlations between the number of CD4⁺ T cells and CD163⁺ and iNOS⁺ macrophages as well as CCR4⁺ and CCR5⁺ cells in both OSCC groups. The mean number of CD8⁺ cells was significantly increased in OSCC M(−) and OLK compared with OSCC M(+) and control group. In OSCC M(+) and OSCC M(−) groups, a negative correlation between the number of CD8⁺ cells and CD163⁺ and iNOS⁺ macrophages was found.Conclusions: The number and co‐localization of lymphocytes and macrophages in OLK and OSCC may indicate that infiltrating cells influence the early and subsequent stage of oral carcinogenesis.

The immunoexpression of the PD-L1 and the number of immune infiltrating cells have been shown to be a significant prognostic factors in various human cancers. Immunohistochemical method was used to examine the immunoexpression of PD-L1 and number of Foxp3+, CD4+, CD8+ cells in 78 cases of oral squamous cell carcinomas (OSCCs): with better prognosis - OSCCBP (n = 37), and with poorer prognosis - OSCCPP (n = 41), and 18 cases of normal mucosa as a control. The immunoexpression of PD-L1 and the mean number of Foxp3+ cells was significantly increased in OSCCPP group in comparison to OSCCBP and control groups. The mean number of CD4+ cells was significantly increased in OSCCPP group in comparison to OSCCBP and control groups. CD8+ cells were significantly more numerous in OSCCBP group in comparison to OSCCPP and control group. In both OSCCPP and OSCCBP groups there were positive significant correlations between number of Foxp3+ and CD4+ cells. We found positive correlations between the immunoexpression of PD-L1 and numbers of Foxp3+ cells, and negative correlation between the immunoexpression of PD-L1 and numbers of CD8+ cells in both OSCCPP and OSCCBP groups. We found also significant positive correlation between immunoexpression of PD-L1 and the number of CD4+ cells in OSCCPP group. In conclusion, our findings support the hypothesis of involvement of Tregs and PD-L1 in OSCC development and progression.

Several lines of evidence indicate that immune cells in the tumor microenvironment play an important role in regulating tumor progression. An immunohistochemical method was used to examine the abundance of natural killer (NK) cells, mucosal dendritic cells (DCs), macrophages, mast cells, and microvessel density in 78 cases of oral squamous cell carcinoma (OSCC): with better prognosis – OSCCBP (n = 37), and with poorer prognosis – OSCCPP (n = 41), and 18 controls. The mean numbers of macrophages and microvessels were significantly higher in the OSCCPP group in comparison to both OSCCBP and control groups. The mean number of NK cells, mast cells and DCs was lower in the OSCCPP group in comparison to the OSCCBP group, but there were no statistically significant differences between mean numbers of NK cells in tested groups. Statistically significant correlations between the number of DCs and NK cells and mast cells, as well as between microvessel density and numbers of macrophages, DCs and mast cells were revealed in both OSCCPP and OSCCBP groups. In conclusion, our findings revealed an association between the number of infiltrating cells and oral cancer prognosis. Moreover, our results suggest that the infiltrating cells (macrophages, Langerhans and mast cells) may be involved in the process of angiogenesis.