Studies of bladder cancer progression

Bladder cancer (BlCa) is the second most common genitourinary cancer, affecting both men and women. Most (70%) cases present at the superficial stage; 20% of these recur with muscle-invasive disease. Major genetic alterations associated with BlCa include: loss/gain in expression or mutations in Retinoblastoma (RB) gene, human epidermal growth factor receptors (HERs), H-ras, p53 and FGFR3. Only p53 mutations are well correlated with invasive BlCa; other changes show variable correlations with disease status. To understand the progression of BlCa, a model of nine human BlCa cell sublines derived from a single parent but differing in in vivo characteristics, has been developed previously. These cells represent a heterogenous population from a single tumour and a model of different stages of BlCa progression, from non-tumourigenic to invasive. Two sublines were selected for further investigation: C3 (non-tumourigenic) and B8 (invasive). These were transfected with green (C3-GSP-2) and red fluorescent reporters (B8-RSP-gck) respectively to investigate the effects of their co-injection in vivo, specifically, promotion of C3 tumour growth by B8 cells. Surprisingly, B8 tumour growth was inhibited by C3 cells in vivo at different cell numbers and proportions of cells injected. Microarray analysis of C3 and B8 cells revealed differential expression of 1367 genes with dramatic differences in the transforming growth factor-ß and integrin-mediated pathways. Gene expression of BMP2, INHBB, FST, NOG, ID4 and TGF- ß1, in the TGF- ß pathway was further analysed with qRT-PCR in all nine sublines. Expression of BMP2 was significantly related to tumourigenic potential (p=0.0238, Mann-Whitney) and INHBB to invasive ability (p=0.0476, Mann-Whitney). The BlCa model did not include a metastatic component. To broaden the model, cell lines were established from an invaded lymph-node (B8-RSP-LN) and a bone-metastasis (B8-RSP-BN) after subcutaneous and intra-cardiac injection of B8-RSP-gck cells. No significant differences were observed in the migratory capability and anchorage-independent colony formation of these metastatic cells compared with B8 cells. Evaluation of expression of the panel of TGF-beta genes (BMP2, INHBB, FST, NOG, ID4 and TGF- ß1) and metastasis-related genes (MMP9, MMP2 and KAI1) indicated that expression of BMP2, FST, ID4 and MMP9 was decreased or lost in the metastatic sublines

Zoledronic Acid Preserves Bone Structure and Increases Survival but Does Not Limit Tumour Incidence in a Prostate Cancer Bone Metastasis Model

Background The bisphosphonate, zoledronic acid (ZOL), can inhibit osteoclasts leading to decreased osteoclastogenesis and osteoclast activity in bone. Here, we used a mixed osteolytic/osteoblastic murine model of bone-metastatic prostate cancer, RM1(BM), to determine how inhibiting osteolysis with ZOL affects the ability of these cells to establish metastases in bone, the integrity of the tumour-bearing bones and the survival of the tumour-bearing mice. Methods The model involves intracardiac injection for arterial dissemination of the RM1(BM) cells in C57BL/6 mice. ZOL treatment was given via subcutaneous injections on days 0, 4, 8 and 12, at 20 and 100 µg/kg doses. Bone integrity was assessed by micro-computed tomography and histology with comparison to untreated mice. The osteoclast and osteoblast activity was determined by measuring serum tartrate-resistant acid phosphatase 5b (TRAP 5b) and osteocalcin, respectively. Mice were euthanased according to predetermined criteria and survival was assessed using Kaplan Meier plots. Findings Micro-CT and histological analysis showed that treatment of mice with ZOL from the day of intracardiac injection of RM1(BM) cells inhibited tumour-induced bone lysis, maintained bone volume and reduced the calcification of tumour-induced endochondral osteoid material. ZOL treatment also led to a decreased serum osteocalcin and TRAP 5b levels. Additionally, treated mice showed increased survival compared to vehicle treated controls. However, ZOL treatment did not inhibit the cells ability to metastasise to bone as the number of bone-metastases was similar in both treated and untreated mice. Conclusions ZOL treatment provided significant benefits for maintaining the integrity of tumour-bearing bones and increased the survival of tumour bearing mice, though it did not prevent establishment of bone-metastases in this model. From the mechanistic view, these observations confirm that tumour-induced bone lysis is not a requirement for establishment of these bone tumours

Molecular profiling of bladder cancer: Involvement of the TGF-Beta pathway in bladder cancer progression

A human bladder cancer model of nine cell sublines derived from the BL17/2 cell line was used to evaluate genes related to disease progression. Molecular profiling of sublines that were non-tumorigenic and invasive in nude mice was performed and identified 1367 differentially-expressed genes. Quantitative real-time PCR analysis of six transforming growth factor-beta (TGF-β) pathway genes using the entire panel of nine cell lines was performed. Bone morphogenetic protein-2 expression was significantly associated with in vivo tumorigenicity of the cell lines (p = 0.0228, Mann-Whitney); inhibin-βB was related to their invasiveness (p = 0.0468, Mann-Whitney). Analysis of conditioned medium showed TGF-β1 production to be significantly associated with the phenotype of the cell line. The study shows the possible involvement of the TGF-β pathway in bladder cancer progression

Text-Guided Graph Neural Networks for Referring 3D Instance Segmentation

This paper addresses a new task called referring 3D instance segmentation, which aims to segment out the target instance in a 3D scene given a query sentence. Previous work on scene understanding has explored visual grounding with natural language guidance, yet the emphasis is mostly constrained on images and videos. We propose a Text-guided Graph Neural Network (TGNN) for referring 3D instance segmentation on point clouds. Given a query sentence and the point cloud of a 3D scene, our method learns to extract per-point features and predicts an offset to shift each point toward its object center. Based on the point features and the offsets, we cluster the points to produce fused features and coordinates for the candidate objects. The resulting clusters are modeled as nodes in a Graph Neural Network to learn the representations that encompass the relation structure for each candidate object. The GNN layers leverage each object's features and its relations with neighbors to generate an attention heatmap for the input sentence expression. Finally, the attention heatmap is used to "guide" the aggregation of information from neighborhood nodes. Our method achieves state-of-the-art performance on referring 3D instance segmentation and 3D localization on ScanRefer, Nr3D, and Sr3D benchmarks, respectively

Monitoring Prostate Tumor Growth in an Orthotopic Mouse Model Using Three-Dimensional Ultrasound Imaging Technique

Prostate cancer (CaP) is the most commonly diagnosed and the second leading cause of death from cancer in males in USA. Prostate orthotopic mouse model has been widely used to study human CaP in preclinical settings. Measurement of changes in tumor size obtained from noninvasive diagnostic images is a standard method for monitoring responses to anticancer modalities. This article reports for the first time the usage of a three-dimensional (3D) ultrasound system equipped with photoacoustic (PA) imaging in monitoring longitudinal prostate tumor growth in a PC-3 orthotopic NODSCID mouse model (n = 8). Two-dimensional and 3D modes of ultrasound show great ability in accurately depicting the size and shape of prostate tumors. PA function on two-dimensional and 3D images showed average oxygen saturation and average hemoglobin concentration of the tumor. Results showed a good fit in representative exponential tumor growth curves (n = 3; r2 = 0.948, 0.955, and 0.953, respectively) and a good correlation of tumor volume measurements performed in vivo with autopsy (n = 8, r = 0.95, P < .001). The application of 3D ultrasound imaging proved to be a useful imaging modality in monitoring tumor growth in an orthotopic mouse model, with advantages such as high contrast, uncomplicated protocols, economical equipment, and nonharmfulness to animals. PA mode also enabled display of blood oxygenation surrounding the tumor and tumor vasculature and angiogenesis, making 3D ultrasound imaging an ideal tool for preclinical cancer research

Histological analysis with tetrachrome stain for effects of zoledronic acid-treatment on normal and tumour-bearing bones.

<p>Tetrachrome stain for differentiation of mineralised and unmineralised bone at 120x magnification Normal (A), or RM1(BM) tumour bearing bones (B–D). (A&B) Untreated, (C) 20 µg/kg ZOL (D) 100 µg/kg ZOL. T = tumour, Arrow = Trabecular bone.</p

Zoledronic acid treatment did not reduce establishment of metastases in RM1(BM) injected mice.

<p>(A) Total number of metastases, (B) bone-metastases, or (C) soft tissue metastases as assessed by one-way ANOVA. Each point represents the number of metastases in an individual mouse, bars indicate the median within the group.</p

Zoledronic acid treatment alters bone volume and the bone surface-area:volume ratio in normal and tumour-bearing bones.

<p>There was no significant change in bone surface area (A) induced by the presence of a tumour or treatment with zoledronic acid. However, bone volume was dramatically reduced in RM1(BM) containing bones and the treatment with ZOL prevented this loss in a dose-dependent manner (B), resulting in lower surface area/volume ratio (increased bone density) compared to normal or tumor-bearing bones (C). Results are based on measurements obtained from CT scans as described in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0019389#s4" target="_blank">Materials and Methods</a> section. Statistical analysis was performed by one-way ANOVA followed by Tukey's post test, * p<0.05, ** p<0.01, *** p<0.001.</p