Reactive oxygen species (ROS) have emerged as cellular signaling molecules and are implicated in metastatic disease by their ability to drive invasion and migration. Here, we define the signaling adaptor protein p130Cas (Crk-associated substrate) as a key redox-responsive molecular trigger that is engaged in highly invasive metastatic bladder tumor cell lines. Endogenous shifts in steady-state hydrogen peroxide (H2O2) that accompany the metastatic phenotype increase p130Cas phosphorylation, membrane recruitment and association with the scaffolding protein Crk, and subsequent Rac1 activation and actin reorganization. Both enzymatic and nonenzymatic scavenging of H2O2 abrogates p130Cas-dependent signaling and the migratory and invasive activity of the metastatic bladder tumor cells. Disruption of p130Cas attenuates both invasion and migration of the metastatic variant (253J-BV). 253J-BV cells displayed an increase in global thiol oxidation and a concomitant decrease in total phosphatase activity, common target proteins of active-site cysteine oxidation. The dependence of phosphatases on regulation of p130Cas was highlighted when depletion of PTPN12 enhanced p130cas phosphorylation and the migratory behavior of a noninvasive parental bladder tumor control (253J). These data show that the metastatic phenotype is accompanied by increases in steady-state H2O2 production that drive promigratory signaling and suggest that antioxidant-based therapeutics may prove useful in limiting bladder tumor invasiveness.
The androgen receptor (AR) is expressed in a subset of prostate stromal cells and functional stromal cell AR is required for normal prostate developmental and influences the growth of prostate tumors. Although we are broadly aware of the specifics of the genomic actions of AR in prostate cancer cells, relatively little is known regarding the gene targets of functional AR in prostate stromal cells. Here, we describe a novel human prostate stromal cell model that enabled us to study the effects of AR on gene expression in these cells. The model involves a genetically manipulated variant of immortalized human WPMY-1 prostate stromal cells that overexpresses wildtype AR (WPMY-AR) at a level comparable to LNCaP cells and is responsive to dihydrotestosterone (DHT) stimulation. Use of WPMY-AR cells for gene expression profiling showed that the presence of AR, even in the absence of DHT, significantly altered the gene expression pattern of the cells compared to control (WPMY-Vec) cells. Treatment of WPMY-AR cells, but not WPMY-Vec control cells, with DHT resulted in further changes that affected the expression of 141 genes by 2-fold or greater compared to vehicle treated WPMY-AR cells. Remarkably, DHT significantly downregulated more genes than were upregulated but many of these changes reversed the initial effects of AR overexpression alone on individual genes. The genes most highly effected by DHT treatment were categorized based upon their role in cancer pathways or in cell signaling pathways (transforming growth factor-?, Wnt, Hedgehog and MAP Kinase) thought to be involved in stromal-epithelial crosstalk during prostate or prostate cancer development. DHT treatment of WPMY-AR cells was also sufficient to alter their paracrine potential for prostate cancer cells as conditioned medium from DHT-treated WPMY-AR significantly increased growth of LNCaP cells compared to DHT-treated WPMY-Vec cell conditioned medium.
Hedgehog signaling regulates Gli transcription factors. Aberrant hedgehog signaling can be oncogenic and drugs that block hedgehog are being tested as anticancer agents. We considered whether hedgehog/Gli signaling may be involved in human bladder transitional cell carcinoma proliferative or invasive behavior.
The role of reactive oxygen species (ROS) in bladder cancer progression remains an unexplored field. Expression levels of enzymes regulating ROS levels are often altered in cancer. A search of publicly available microarray data reveals that expression of mitochondrial manganese superoxide dismutase (Sod2), responsible for the conversion of superoxide (O(2)(-)) to hydrogen peroxide (H(2)O(2)), is consistently increased in high-grade and advanced-stage bladder tumors. We aimed to identify the role of Sod2 expression and ROS in bladder cancer. Using an in vitro human bladder tumor model we monitored the redox state of both nonmetastatic (253J) and highly metastatic (253J B-V) bladder tumor cell lines. 253J B-V cells displayed significantly higher Sod2 protein and activity levels compared to their parental 253J cell line. The increase in Sod2 expression was accompanied by a significant decrease in catalase activity, resulting in a net increase in H(2)O(2) production in the 253J B-V cell line. Expression of the prometastatic and proangiogenic factors matrix metalloproteinase 9 (MMP-9) and vascular endothelial-derived growth factor (VEGF), respectively, was upregulated in the metastatic line. Expression of both MMP-9 and VEGF was shown to be H(2)O(2)-dependent, as removal of H(2)O(2) by overexpression of catalase attenuated their expression. Similarly, expression of catalase effectively reduced the clonogenic activity of 253J B-V cells. These findings indicate that metastatic bladder cancer cells display an altered antioxidant expression profile, resulting in a net increase in ROS production, which leads to the induction of redox-sensitive protumorigenic and prometastatic genes such as VEGF and MMP-9.
Extended biopsy schemes are now the standard of care for detection of prostate cancer. Submitting biopsy cores individually raises the cost of pathologic evaluation significantly while important prognostic information is lost when the samples are bundled into fewer containers. We devised a protocol for bundling biopsy cores to reduce the cost while maintaining our ability to identify important biopsy features.
Complete eversion of the urinary bladder is a rare problem that presents a serious management challenge. Currently no standard treatment recommendations exist for management. We describe our experience with bladder eversion and present an algorithm for treatment.
Prostatic adenocarcinoma is the most frequently diagnosed cancer in American men. Tumor Gleason grade and stage provide extremely valuable prognostic information and play an important role in therapeutic decision making and patient counseling. A biopsy or radical prostatectomy specimen revealing carcinoma extending into extraprostatic tissue permits a T3 classification. This is most easily recognized, particularly in a needle biopsy, when tumor is seen to invade the adipose tissue. The existence of intraprostatic adipose tissue is somewhat controversial. To investigate this, formalin-fixed paraffin-embedded whole-mount radical prostatectomy specimens from 427 patients with adenocarcinoma were evaluated for intraprostatic adipose tissue. It was defined as any collection of adipocytes amid or internal to the most peripheral glands. The amount, anatomic location, and relationship to normal structures were also recorded. Intraprostatic adipose tissue was identified in 17 (3.98%) of cases. It consisted of small microscopic foci composed of 5 to 20 adipocytes. In 13 cases, the fat was intimately associated with benign glands. In another 2 cases, it was associated with small nerves, and in 2 cases was random with no specific localization. Intraprostatic adipose tissue was located in the peripheral zone in 15 cases and in the central zone in 2. Intraprostatic adipose tissue, although uncommon, does exist. Therefore, caution must be exercised in diagnosing extraprostatic extension based only upon identification of fat invasion, especially in a needle biopsy. The small size of foci of adipose tissue and its admixture with benign glands are useful morphologic clues in distinguishing it from extraprostatic fat.
To characterize the clinicopathologic features of patients who developed pubovesical fistula (PVF) after treatment of prostate cancer and to identify some possible methods of reducing the incidence of this rare complication for which no well-established guidelines exist.
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