Background Obesity is associated with prostate cancer aggressiveness and mortality. of adipocytes plus stromal-vascular fraction (SVF) as opposed to SVF alone. MMP2 activity was higher for PP when compared to VIS adipose tissue. When PC-3 cells were stimulated with CM from PP adipose tissue explants, increased proliferative and migratory capacities were observed, but not in the presence of SVF. Conversely, when LNCaP cells were stimulated with PP explants CM, we found enhanced motility despite the inhibition of proliferation, whereas CM derived from SVF increased both cell proliferation and motility. Explants buy INCB39110 culture and using adipose tissue of PP origin are most effective in promoting proliferation and migration of PC-3 cells, as respectively compared with SVF culture and using adipose buy INCB39110 tissue of VIS origin. In LNCaP cells, while explants CM cause increased migration compared to SVF, the use of PP adipose tissue to generate CM result in the increase of both cellular proliferation and migration. Conclusions Our findings suggest that the PP depot has the potential to modulate extra-prostatic tumor cells’ microenvironment through increased MMPs activity and to promote prostate cancer cell survival and migration. Adipocyte-derived factors likely have a relevant proliferative and motile role. Keywords: Adipose tissue, Cell line, Cell proliferation, Cell tracking, Obesity, Periprostatic, Prostate cancer Background In recent years substantial evidence has been provided for the linkage between adipose tissue dysfunction and cancer progression [1,2]. Excess accumulation of adipose tissue corresponds by definition to obesity, which has been associated with prostate cancer aggressiveness [3,4]. In prostate cancer, the extra-capsular extension of cancer cells into the periprostatic (PP) fat is a pathological factor related with worst prognosis [5]. It is now well established that the interactions between non-tumor cells in the microenvironment and the tumor cells are decisive of whether cancer cells progress towards metastasis or whether Rabbit Polyclonal to VANGL1 they remain dormant [6]. Prostate cancer cells generated within prostatic acini frequently infiltrate and even surpass the prostatic capsule, therefore interacting with the surrounding PP adipose tissue. Previous work showed that such adipose tissue has the potential to modulate prostate cancer aggressiveness, through the increased production of adipokines, namely interleukin 6 (IL-6) [7]. Moreover, a recent report showed an association of PP adipose tissue thickness with prostate cancer severity [8]. Different studies have demonstrated the critical influence of adipose tissue-derived factors in cancer cells [9-11], including prostate tumor cells [12-14]. Together, these reports indicate that factors produced by adipose tissue, particularly adipocytes may stimulate the progression of cancer cells. However, to our knowledge, the influence of PP adipose tissue-derived factors on prostate cancer cells has not been exploited. Noteworthy, we previously observed that prostate cancer induced the increase of PP buy INCB39110 adipose metabolic activity, promoting a favorable environment for aggressive tumor biology [15]. To address these issues, we first studied the gelatinolytic profile of PP whole adipose tissue and its respective stromal-vascular fraction. Next, we used PP adipose tissue-derived conditioned medium to analyze in vitro its influence in proliferation and migration of prostate cancer cells. Methods Patients and collection of human PP adipose tissue Men diagnosed with clinically localized prostate cancer or nodular prostatic hyperplasia (BPH) and eligible for retropubic radical prostatectomy or prostate surgery of nodular hyperplasia, without other major co-morbidities, were included in this study after informed consent agreement. The project was approved by the ethics committees of the participating Hospitals. Human anterior-lateral PP and pre-peritoneal visceral (VIS) samples of adipose tissue were collected during surgery and immediately processed. Adipose tissue primary cultures and preparation conditioned media (CM) PP and VIS adipose tissue fragments were processed to primary whole adipose tissue (explants) cultures using a modified protocol from Thalmann et al. [16]. Briefly, after incubation of explants (0.3 g/mL) for 16 hours in DMEM/F12 (Gibco) medium, supplemented with biotin 16 M (Sigma Aldrich), panthotenate 18.