However, specificity improved when combinations of different biomarkers were evaluated, especially among SCC cases [31]. In our study only a single non-invasive technique was employed and the results confirm

that cutaneous swabs cannot be utilized as a single method for epidemiological studies on HPV associated skin cancer. Immunohistochemistry analysis p16INK4a immunostaining Immunohistochemistry detected p16INK4a expression in 33 of 35 (94,2%) tumor samples. In particular a higher score (≥ 30% of p16INK4a positive dysplastic keratinocytes) was detected in 8 cases (Table 1 and Figure 3). Absent or weak p16INK4a expression was documented in rare cells see more of few perilesional skin samples (Figure 3). Figure 3 Immunostaining patterns of p16 Ink4a . Gemcitabine solubility dmso BCC (A) with high number of p16Ink4a positive dysplastic keratinocytes and normal skin (B) with rare positive

normal keratinocytes. Sections were counterstained with haematoxylin. Magnification A (20×) and B (10×). These data contrast with those showing that an inactivation of p16INK4a is commonly associated with more malignant features in many tumors [31], including BCC [32–37]. However other reports stated a strong p16INK4a mRNA expression in BCC skin [38–40]. Eshkoor et al. [39] found a significant protein and mRNA expression in BCC cells when compared with normal skin tissue. In particular the samples they tested were paraffin-embedded skin BCC as our samples. Indeed conflicting results could be attributed to different methods used, which need

further optimization of experimental conditions. SCH 900776 in vivo Furthermore, there appears to be a strong relationship between the level of invasiveness and expression of p16INK4a. Svensson et al. [40] showed that p16INK4a expression is associated with a highly invasive BCC subtype with infiltrative growth patterns. In the mean time the results of Conscience et al. [38] contradict those of Svensson et al. [40], as they did not observe any difference in the expression of p16INK4a among different histological types of carcinoma suggesting that p16INK4a expression does not Flucloronide correlate with malignancy or proliferation. On the contrary the p16INK4a over-expression was found significantly associated with the BCC location on sun-exposed areas. Our data did not evidence such association and are more consistent with those of Eshkoor et al. [39] and Svensson et al. [40]. Akt 1/2 immunostaining Immunohistochemistry detected pAkt1 expression in 30 out of 32 (93,7%) tumor samples (Table 1 and Figure 4). with only 2 cases showing rare positive cells. Most of the positive cells showed signal in the cytoplasm and in the nucleus, suggesting that pAkt properly translocates in the nucleus to exert its activity. Thus the PI3K ⁄Akt pathway is activated in BCC examined in our study. Figure 4 Immunostaining patterns of pAkt and Akt2.