miR-29b enhances prostate cancer cell invasion independently of MMP-2 expression.

BACKGROUND: The ability to metastasize is one of the most important characteristics of neoplastic cells. An imbalance between the action of some matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs drives the invasion process. Some studies have suggested that MMP-2 is involved in metastasis, while other studies have reported that collagen production by cancer cells might also contribute to motility. However, decreased expression of microRNA-29b (miR-29b), which may control MMP-2 and collagen gene expression, has been shown in prostate cancer (PCa). The objectives of the present study were to clarify whether MMP-2 as well as collagens I and III (encoded by COL1A1 and COL3A1, respectively) are controlled by miR-29b and to determine whether metastasis is altered by this relationship. METHODS: PCa DU145 and PC-3 cells were transfected with 100 µL of OPTI-MEM I containing 100 nmol of miR-29b (or its inhibitor) along with 1.5 µL of lipofectamine. Positive and negative controls were prepared using the same protocol. MMP-2, COL1A1 and COL3A1 messenger RNA (mRNA) levels were evaluated via real-time polymerase chain reaction (qRT-PCR). For qRT-PCR, 6 × 104 cells were used. Invasion studies were conducted with Matrigel assays, which simulate invasion of the extracellular matrix by neoplastic cells. After transfection of 3 × 104 cells, invasion was allowed to proceed for 48 h. Invasive cells were counted under an optical microscope. Each experiment was performed in triplicate. RESULTS: MMP-2 mRNA was not expressed in DU145 cells after transfection with miR-29b. After transfection of cells with the miR-29b inhibitor, COL1A1 (p = 0.02) and COL3A1 (p = 0.06) mRNA expression was increased in DU145 cells, and a large number of transfected DU145 and PC3 cells invaded the Matrigel membrane. CONCLUSIONS: In vitro studies showed that reducing the amount of miR-29b may lead to higher PCa cell invasion via a process that is independent of MMP-2. Collagen expression, controlled by miR-29b, may facilitate this motility process. Thus, the present study suggests that collagen production plays an active role in metastasis control and restoration of miR-29b levels may decrease metastasis. Altogether, these findings support further exploration of drug therapy targeting this aspect of the metastasis circuit.

miR-618: possible control over TIMP-1 and its expression in localized prostate cancer.

BACKGROUND: The imbalance between the action of the tissue inhibitors of matrix metalloproteinases (TIMPs) and the matrix metalloproteinases (MMPs) is one component of metastasis physiology. TIMP-1 overrides MMP-9 activity in cancer and might be regulated by miR-618. The aims of this study were to clarify whether TIMP-1 expression is modified by miR-618 and to clarify the effect of miR-618 expression on the invasion of prostate cancer cells. We also studied miR-618 expression in surgical specimens of patients with localized prostate cancer submitted to open radical prostatectomy. METHODS: After transfection of miR-618 or its antagonist in DU145 cells, qRT-PCR for TIMP-1/MMP-9 and both ELISA and zymography for MMP-9 were performed. Total miRNA was extracted from surgical specimens of PCa, and miR-618 expression was examined for correlations with Gleason score, pathological status and biochemical recurrence. RESULTS: DU145 cells transfected with miR-618 had a 76% reduction in TIMP-1 expression relative to control cells (p = 0.003). miR-618 inhibition reduced MMP-9 expression by 31% (p = 0.032) and MMP-9 absorbance evaluated with ELISA assay (p = 0.06).Zymography suggested higher MMP-9 activity in DU145 cells transfected with miR-618 than those transfected with miR-618 inhibitor, but the difference was not significant (p = 0.55). However, miR-618 expression was lower in surgical specimens of patients with Gleason score > 7 (p = 0.08) and more advanced disease (p = 0.07). CONCLUSIONS: In vitro, miR-618 overexpression decreases TIMP-1 and miR-618 inhibition decreases MMP-9, suggesting that miR-618 might be an oncomiR. However, the analysis of clinical samples of localized prostate cancer revealed an inconsistent pattern, as increased miR-618 expression was associated with lower Gleason score and pathological status. Further studies are needed to address whether miR-618 is a context-dependent miRNA.

Impact of Camera Deviation on Penile Curvature Assessment Using 2D Pictures.

BACKGROUND: An accurate curvature assessment (CA) is required in the decision-making process for patients with Peyronie's disease. In-office CA following induced erection is the gold standard for CA, although penile photography is commonly used due to its convenience. Camera deviations during 2D image acquisition might affect CA accuracy. AIM: To investigate the impact of camera angle deviations on CA. METHODS: 2D pictures were taken from 5 models with a known uniplanar curvature (40°, 45°, 60°, 90°, and 120°). The model was kept on a fixed point and the camera was rotated around it. Pictures were taken with every 10° increase in camera deviation from the optimal position. The camera rotated to a maximum of 90° deviation in both the vertical and horizontal planes. The pictures were analyzed by 2 different urologists using a goniometer. The expected apparent curvature (AC) and the corresponding picture assessment error (PAE = AC - real model curvature) were also calculated for each picture using trigonometry principles. MAIN OUTCOME MEASURE: Assessing PAE magnitude and patterns was our primary outcome. Secondary outcomes were intraobserver, interobserver, and observer-AC intraclass correlation coefficient (ICC). RESULTS: 100 pictures were analyzed. Intraobserver reliability was high (ICC = 0.99) for both urologists. Interobserver and observer-AC correlation were also high (ICC = 0.996 and ICC = 0.992, respectively). When the camera rotated in the horizontal axis, the PAE underestimated the curvature for models with curvatures smaller than 90° and overestimated the reading of the 120° model. When the camera rotated in the vertical axis, PAE had an inverse effect. The PAE showed a tendency to increase exponentially with higher deviation, reaching almost 100% for a deviation of 80°. Nevertheless, analyzing its magnitude regardless of the curvature, PAE was always <5% for camera deviations of 0-20°. CLINICAL IMPLICATIONS: If using picture-based CA, clinicians should attempt to take a picture perpendicular to the curvature plane for the most accurate measurement in degrees. Many clinicians request that patients take 3 pictures in a standard fashion (craniocaudal, lateral, and frontal), and if this technique is to be used, an extra picture is recommended. STRENGTH & LIMITATIONS: In our controlled environment, we were able to isolate CA errors due to camera angles from other confounders such as erection hardness. As a consequence, however, our results cannot be easily generalized. CONCLUSION: PAE due to non-optimal camera position is a complex phenomenon that affects CA depending on the rotation axis and the degree of penile curvature. Nevertheless, PAE is always <5% for camera deviations of 0-20°. Nascimento B, Cerqueira I, Miranda EP, et al. Impact of Camera Deviation on Penile Curvature Assessment Using 2D Pictures. J Sex Med 2018;15:1638-1644.

MMP9 overexpression is associated with good surgical outcome in children with UPJO: Preliminary results.

BACKGROUND: Ureteropelvic junction obstruction (UPJO) diagnosed prenatally occurs in 1:150 - 1:1200 pregnancies. Although many studies investigating the molecular changes of this obstructed segment have been performed, the underlying mechanisms are still unclear. The role of extracellular matrix (ECM) components remains controversial, and the investigations in the field of ECM changes, might help the better understanding of the pathogenesis of this common condition. The aim of the present study was to investigate for the first time in the literature whether MMP9 and its specific inhibitors, TIMP1 and RECK, are expressed in a reproducible, specific pattern in UPJ. METHODS: UPJO specimens were obtained from 16 children at the time of dismembered pyeloplasty due to intrinsic UPJ stenosis. Expression levels of the three genes (MMP9, TIMP1 and RECK) were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). Then correlated the expression levels of the genes according to grade study population that was divided in 2 categories according to Society of Fetal Urology classification, grade 3 (moderate) and 4 (severe). For DTPA we subdivided the childrens in 2 groups, obstructive (T 1/2 more than 20 min) and partial obstructive (T 1/2 between 10 and 20 min) and success in a surgery was defined as decrease in T 1/2 to less than 20 min, absence of symptoms, improving renal function and decreasing dilatation on successive exams. RESULTS: MMP9 was underexpressed and TIMP1 and RECK were overexpressed in children with obstructive DTPA but the differences were not statistically significant. Overexpression of MMP9 was higher among patients with severe grade of UPJ compared to those with moderate grade. Surprisingly expression levels of MMP-9 was three times higher in children who were successfully treated by surgery (n = 10) (p = 0.072), so those who were followed for at least 1 year after surgery and remained with improvement in renal function and decreasing dilation on intravenous urogram and TIMP-1 was underexpressed in 100 % of this cases (p = 0.00). CONCLUSIONS: We showed an increase in expression of MMP9 and a decrease in expression of TIMP1 in children who improving renal function and decreasing dilation after surgery. We believe that the higher expression of MMP9 in these cases can reflect an increase in degradation and remodeling process that could be used as a marker for surgical outcome.

Recurrent Penile Fracture-Case Report and Alternative Surgical Approach.

INTRODUCTION: Penile refracture is an exceedingly rare event, with very few published studies. To the best of our knowledge, this is the first documented case in the literature of penile fracture with 3 same-site recurrences. AIMS: To describe the case of a 25-year-old Caucasian man with recurrent penile fracture ultimately treated with resuture and patch reinforcement. METHODS: Patient history (clinical and surgical) and literature review. RESULTS: After the 3rd same-site recurrence, patch reinforcement over the sutured area was performed. The patient had an uneventful recovery and no recurrences to date. CONCLUSION: There is no evidence indicating the superiority of non-absorbable sutures. Bovine pericardium reinforcement over the sutured area was used to minimize the chance of another recurrence. More studies are necessary to investigate its safety and efficacy in this scenario. Nascimento B, Guglielmetti GB, Miranda EP, et al. Recurrent Penile Fracture-Case Report and Alternative Surgical Approach. Sex Med 2018;6:263-266.