HR-positive/HER2-negative breast cancer arising in patients with or without BRCA2 mutation: different biological phenotype and similar prognosis.

Background: BRCA2 plays a key role in homologous recombination. However, information regarding its mutations in Chinese patients with breast cancer remains limited. Objectives: This study aimed to assess the clinicopathological characteristics of BRCA2 mutation breast cancer and explore the mutation's effect on hormone receptor (HR)-positive/human epidermal growth factor receptor 2 (HER2)-negative breast cancer survival in China. Design: This hospital-based cohort study prospectively included 629 women with breast cancer diagnosed from 2008 to 2023 at Zhejiang Cancer Hospital in China. Methods: We compared the clinicopathological characteristics and metastatic patterns and analysed the invasive disease-free survival (iDFS), distant relapse-free survival (DRFS) and first-line progression-free survival (PFS1) of patients with HR-positive/HER2-negative breast cancer according to BRCA2 mutations. Results: Among the 629 patients, 78 had BRCA2 mutations (12.4%) and 551 did not (87.6%). The mean age at diagnosis was lower in the BRCA2 mutation breast cancer group than in the non-mutation breast cancer group (38.91 versus 41.94 years, p = 0.016). BRCA2 mutation breast cancers were more likely to be lymph node-positive than non-mutation breast cancers (73.0% versus 56.6%, p = 0.037). The pathological grade was higher in 47.1% of BRCA2 mutation breast cancers than in 29.6% of non-mutation breast cancers (p = 0.014). The proportions of patients with BRCA2 mutations who developed contralateral breast cancer (19.2% versus 8.8%, p = 0.004), breast cancer in the family (53.8% versus 38.3%, p = 0.009) and ovarian cancer in the family (7.6% versus 2.4%, p = 0.022) were higher than those of patients without the mutation. The median follow-up time was 92.78 months. Multivariate analysis showed that BRCA2 mutation was not associated with poorer iDFS [hazard ratio = 0.9, 95% confidence interval (CI) = 0.64-1.27, p = 0.56] and poorer distant relapse-free survival (DRFS) (hazard ratio = 1.09, 95% CI = 0.61-1.93, p = 0.76). There was no significant difference between the two groups with regard to metastatic patterns in the advanced disease setting. In the first-line metastatic breast cancer setting, PFS1 expression was broadly similar between the two groups irrespective of chemotherapy or endocrine therapy. Conclusion: HR-positive/HER2-negative breast cancer with BRCA2 mutations differs from those without mutations in clinical behaviour and reflects more aggressive tumour behaviour. Our results indicate that BRCA2 mutations have no significant effect on the survival of Chinese women with HR-positive/HER2-negative breast cancer.

Giant dumbbell-shaped prostatic cystadenoma presenting as pelvic and scrotal mass.

This study reports the case of an elderly man with a large tumour of the pelvic cavity and scrotum which was once diagnosed as a prostate cyst. Imaging studies considered the source of the tumour to be prostate, and the tumour was ultimately diagnosed by confirmed tissue expression of prostate specific antigen (PSA) and prostate acid phosphatase (PSAP) after surgery. This is the first report about dumbbell-shaped prostatic cystadenoma with invasive growth and even urethral damage, but there was no evidence of clear malignancy. Early diagnosis and treatment are crucial in such kinds of diseases.

A significant response to a combination of trastuzumab and vinorelbine in HER2-negative metastatic breast cancer with HER2 V777L mutation.

Metastatic breast cancer (MBC) is the most life-threatening disease in women worldwide. HER2-mutated breast carcinoma has been reported to benefit from HER2-targeted tyrosine kinase inhibitors recently. Here, we presented a heavy pretreated and harbored HER2 V777L mutation de novo stage IV Luminal B (HER2 unamplified) breast cancer patient who achieved an unexpected good response to trastuzumab combined with vinorelbine therapy. Although HER2-unamplified MBC patients do not regularly benefit from anti-HER2 target therapy, HER2 V777L mutation detected by next-generation sequencing from ctDNA may present as a predictive biomarker for anti-HER2-based strategy therapy in HER2-negative MBC patients.

Meta-analysis of microRNAs as biomarkers for muscle-invasive bladder cancer.

Bladder cancer is the most common cancer of the urinary tract. A quarter of bladder cancer patients presenting with muscle-invasive bladder cancer (MIBC) suffer significant morbidity and succumb to the disease. MicroRNA (miRNA) from tissue, urine or blood samples of MIBC patients have been demonstrated to differ from healthy individuals, and possibly have diagnostic value. The aim of the present meta-analysis was to access the overall diagnostic accuracy comprehensively and quantitatively. Systematic searching in PubMed, Web of Science, Embase and Chinese National Knowledge Infrastructure database was conducted. The pooled sensitivity, specificity, positive and negative likelihood ratios (PLR and NLR) and diagnostic odds ratio (DOR) were calculated via the random effects model to evaluate the overall test performance. Deeks' funnel plot asymmetry test was used to test the publication bias. A total of 10 studies were included in the meta-analysis, with a total of 577 patients and 412 controls. The pooled sensitivity and specificity were 0.78 [95% confidence interval (CI), 0.69-0.86] and 0.77 (95% CI, 0.72-0.81), respectively. The pooled PLR was 2.9 (95% CI, 2.1-3.8), the NLR was 0.31 (95% CI, 0.27-0.35), the DOR was 7 (95% CI, 4-13) and the pooled AUC was 0.80 (95% CI, 0.69-0.87). In conclusion, the current miRNA assays support their use as markers for MIBC diagnosis.

CTA combined with CT perfusion for assessing the efficacy of anti-angiogenic therapy in rabbit VX2 tumors.

RATIONALE AND OBJECTIVES: The aim of this study was to validate the feasibility of assessing the efficacy of antiangiogenic therapy on VX2 tumors using three-dimensional computed tomographic (CT) angiography (CTA) combined with CT perfusion. MATERIALS AND METHODS: Forty rabbits with VX2 tumors were randomly assigned to four groups according to different doses of antiangiogenic drug, which were administered intraperitoneally daily for 14 days. In each group, 10 animals were scanned using three-dimensional CTA and CT perfusion on days 1 and 2 after the latest administration of the drug. Tumor masses were sectioned, stained by immunohistochemistry, and processed for correlation between CT imaging and histology. RESULTS: The numbers of new tumor vessels from CTA were significantly different among the four groups (P < .001). As the dose of the drug increased, blood flow and blood volume on CT perfusion increased linearly, but the mean transit time and permeability surface-area product decreased linearly (P < .001). Immunohistochemical analyses showed that microvascular density decreased, while both luminal vascular number and mature vessel number increased linearly as the drug dose increased (P < .001). CT manifestations were correlated well with histologic findings (P < .05). CONCLUSIONS: It is feasible to assess the efficacy of antiangiogenic therapy on VX2 tumors using three-dimensional CTA combined with CT perfusion. Three-dimensional CTA can display the morphologic changes of tumor vessels, while CT perfusion can predict the functional changes of tumor vessels after antiangiogenic therapy.

Slit-Robo GTPase-activating proteins are differentially expressed in murine dorsal root ganglia: modulation by peripheral nerve injury.

The Slit-Robo GTPase-activating proteins (srGAPs) play an important role in neurite outgrowth and axon guidance; however, little is known about its role in nerve regeneration after injury. Here, we studied the expression of srGAPs in mouse dorsal root ganglia (DRG) following sciatic nerve transection (SNT) using morphometric and immunohistochemical techniques. Reverse transcriptase polymerase chain reaction and Western blot analysis indicated that srGAP1 and srGAP3, but not srGAP2, were expressed in normal adult DRG. Following unilateral SNT, elevated mRNA and protein levels of srGAP1 and srGAP3 were detected in the ipsilateral relative to contralateral L(3-4) DRGs from day 3 to day 14. Immunohistochemical results showed that srGAP1 and srGAP3 were largely expressed in subpopulations of DRG neurons in naïve DRGs. However, after SNT, srGAP3 in neurons was significantly increased in the ipsilateral relative to contralateral DRGs, which peaked at day 7 to day 14. Interestingly, DRG neurons with strong srGAP3 labeling also coexpressed Robo2 after peripheral nerve injury. These results suggest that srGAPs are differentially expressed in murine DRG and srGAP3 are the predominant form. Moreover, srGAP3 may participate in Slit-Robo signaling in response to peripheral nerve injury or the course of nerve regeneration.

A comparison of 3D-CTA and 4D-CE-MRA for the dynamic monitoring of angiogenesis in a rabbit VX2 tumor.

PURPOSE: To compare three-dimensional computed tomography angiography (3D-CTA) and four-dimensional contrast-enhanced magnetic resonance angiography (4D-CE-MRA) for the in vivo monitoring of tumor angiogenesis. MATERIALS AND METHODS: VX2 tumors were implanted into the right thigh muscle of 30 New Zealand white rabbits. The animals were randomly assigned to 5 groups, which, respectively, were scanned by 3D-CTA and 4D-CE-MRA on day 4, 7, 10, 13, or 16 after tumor implantation. After scanning, tumors were resected and processed for conventional histology and CD-31 immunohistochemistry. Tumor volume measurements derived from CT and MR imaging were compared with histopathological data. The minimum tumor diameter and the number of new tumor blood vessels detectable by 3D-CTA and 4D-CE-MRA were also compared. RESULTS: There were no significant differences in the tumor volume measurements derived from CT, MR, and histological analysis. The minimum diameter of tumor vessels detectable by 3D-CTA (0.68 ± 0.07 mm) was significantly less than that by 4D-CE-MRA (0.85 ± 0.12 mm) (P=0.005). The number of tumor vessels detected by each imaging method was not significantly different until day 13 after implantation, when 3D-CTA detected a greater number (P<0.001). The morphologic process of tumor angiogenesis was demonstrated dynamically by 3D-CTA and 4D-CE-MRA in vivo. CONCLUSIONS: Tumor angiogenesis can be dynamically monitored in vivo by 3D-CTA and 4D-CE-MRA. Of the two methods, 3D-CTA has better spatial resolution, but 4D-CE-MRA allows temporal resolution of tumor angiogenesis.

Studies of pathology and VEGF expression in rabbit cerebrospinal fluid metastasis: application of dynamic contrast-enhanced MRI.

OBJECTIVE: The objective was to analyze the correlation of dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) with vascular endothelial growth factor (VEGF) protein expression and to assess the potential application of DCE-MRI to the rabbit cerebrospinal fluid (CSF) metastasis model. METHODS: Thirty New Zealand rabbits were divided into experimental and control groups. In the experimental group, VX2 tumor cells were injected into the subarachnoid space at the plane of cisterna magna in 24 rabbits. In the control group, physiological saline was injected into the subarachnoid space at the plane of cisterna magna in six rabbits. DCE-MRI was performed at multiple time points, and several pharmacokinetic parameters, including K(trans), K(ep) and V(e), were calculated. Also, VEGF levels in plasma and CSF were evaluated by enzyme-linked immunosorbent assay prior to DCE-MRI examination. After DCE-MRI examination, the rabbits were sacrificed, and the corresponding tumor specimens were harvested. Hematoxylin-eosin staining and VEGF immunohistochemical staining were carried out, and VEGF expression in the specimens was evaluated by the immunohistochemical scoring system. RESULTS: Vascular endothelial growth factor positive staining was localized in the cytoplasm and cell membranes of tumor cells, as well as in a subset of epithelial cells. Both VEGF immunohistochemical scores and VEGF expression in CSF and plasma exhibited positive correlations with K(trans) and K(ep) values as demonstrated by rank correlation statistical analysis. CONCLUSIONS: Vascular endothelial growth factor expression in plasma and CSF in the CSF metastasis model was higher than in normal tissues. Therefore, DCE-MRI reliably indicated VEGF expression in the rabbit CSF metastasis model.

MicroRNAs 144, 145, and 214 are down-regulated in primary neurons responding to sciatic nerve transection.

MicroRNAs (miRNAs) play an important role in the development, differentiation, proliferation, survival, and oncogenesis of cells and organisms including nervous system. However, the role of miRNAs in primary neurons of dorsal root ganglion (DRG) after injury was not clear. In this study, a miRNA microarray analysis was performed, and a total of 21 miRNAs were found to be down-regulated following unilateral sciatic nerve transection. The miR-144, miR-145, and miR-214 were further validated using quantitative reverse transcriptase PCR (qRT-PCR). Moreover, in situ hybridization (ISH) experiments using locked nucleic acid (LNA)-modified DNA oligonucleotide probes verified that miR-144, miR-145, and miR-214 were expressed in primary neurons of DRG and down-regulated following sciatic nerve transection. Predictions of potential miRNA targets involved were identified by performing a bioinformatics analysis. These predictions were tested using miRNA luciferase reporter vectors, with Robo2 and srGAP2 evaluated as the potential targets of miR-145 and miR-214, respectively. The role of miR-145 in cultured primary neurons was also investigated, and the result found that miR-145 miR-145 inhibited neurite growth and down-regulated Robo2 expression. Finding from this study suggested that miRNAs of DRG can mediated the course of regeneration including through Slit-Robo-srGAP signaling pathway after injury.

Combination of vascular endothelial growth factor antisense oligonucleotide therapy and radiotherapy increases the curative effects against maxillofacial VX2 tumors in rabbits.

PURPOSE: To study the effects of combination of vascular endothelial growth factor (VEGF) antisense oligonucleotide therapy and radiotherapy on maxillofacial VX2 tumors in rabbits. METHODS: We used 24 New Zealand white rabbits as a model to induce maxillofacial VX2 tumor. The rabbits were randomly divided into the following 4 groups: radiotherapy group (group A), treated with 16 Gy of radiotherapy; VEGF antisense oligonucleotide treatment group (group B), treated with an injection of 150 µg of VEGF antisense oligonucleotide into the local tumor; VEGF antisense oligonucleotide combined with radiotherapy group (group C), treated with an injection of 150 µg of VEGF antisense oligonucleotide into the local tumor immediately after 16 Gy of radiotherapy; and control group (group D), treated with an injection of 300 µl 5% aqueous glucose solution into the local tumor. On days 3 and 14 after treatment, dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) was performed to calculate maximal enhancement ratio (MER), slope of enhancement (SLE), and tumor volume change. Rabbits were killed on day 14 to obtain samples for pathological examination and immunohistochemical staining for VEGF. RESULTS: In group C, tumor volume was significantly reduced on day 14 after treatment, and the difference was statistically different as compared to that before treatment, on day 3 after treatment and other groups (P < 0.01). Values of both MER and SLE after treatment were significantly lower than the values before treatment (P < 0.05). Pathological specimen revealed tumor cell edema, bleeding, necrosis, vascular wall thickening and occlusion, and decreased VEGF expression. The immunohistochemical score (IHS) of group C was significantly different from groups A and D respectively (P < 0.05). CONCLUSION: Injecting the tumor with VEGF antisense oligonucleotide immediately after radiotherapy can enhance the curative effect on rabbit maxillofacial VX2 tumor, and DCE-MRI can serve as a reliable technique for in vivo monitoring.

Expression of HGF/c-Met is dynamically regulated in the dorsal root ganglions and spinal cord of adult rats following sciatic nerve ligation.

Hepatocyte growth factor (HGF) and its receptor c-Met play pivotal roles in post-traumatic regeneration of the nervous system. However, following peripheral nerve injury, the role and regulation of the HGF/c-Met system is less clear. Therefore, using a sciatic nerve ligation (SNL) model, spatiotemporal changes in HGF and c-Met expression were detected in the dorsal root ganglions (DRGs) and lumbar spinal cords of adult rats. HGF expression following SNL was found to be significantly decreased in ipsilateral L4-L5 DRGs from day 3 to day 14, with the lowest levels of expression detected on days 5 and 7. In contrast, no significant change in HGF expression was detected in the lumbar spinal cords. c-Met expression in ipsilateral L4-L5 DRGs and within the ipsilateral dorsal horn was found to be significantly up-regulated following SNL, particularly from day 5 to day 14, with peak levels of expression detected on days 7 and 14. In contrast, c-Met levels following SNL consistently remained stable in the spinal ventral horn. These findings suggest that the HGF/c-Met system is spatiotemporally regulated by a unique pattern of signaling pathways induced by peripheral nerve injury, and these pathways have a role in promoting the survival of injured neurons, especially adult DRG sensory neurons.

Slit1 promotes regenerative neurite outgrowth of adult dorsal root ganglion neurons in vitro via binding to the Robo receptor.

Secreted Slit proteins have previously been shown to signal through Roundabout (Robo) receptors to negatively regulate axon guidance and cell migration. During vertebrate development, Slit proteins have also been shown to stimulate branching and elongation of sensory axons and cortical dendrites. In this study, Slit1/Robo2 mRNA and protein expressions were detected in adult rat dorsal root ganglion (DRG) and in cultured DRG neurons. Treatment of both models with recombinant, soluble Slit1 protein was found to promote neurite outgrowth and elongation. In contrast, treatment with a recombinant human Robo2/Fc chimera inhibited neurite outgrowth and elongation. When adult DRG and cultured DRG neurons were pretreated with soluble recombinant human Robo2/Fc chimera, neurite outgrowth and elongation was not induced. These findings indicate that Slit1/Robo2 signaling may have a role in regulating peripheral nerve regeneration.

[Comparison between Gd-DTPA-enhanced MRI and USPIO-enhanced MRI in diagnosing benign and metastatic cervical lymph nodes in rabbit models].

OBJECTIVE: To compare the diagnostic value of gadolinium diethylenetriaminepenta-acetic acid (Gd-DTPA)-enhanced MRI with ultrasmall superparamagnetic iron oxide (USPIO)-enhanced MRI in differentiating reactive hyperplastic lymph nodes from metastatic lymph nodes in rabbit models. METHODS: Reactive hyperplastic cervical lymph node model was established in 18 rabbits as hyperplasia group, and tumor-bearing lymph node model was established in another 18 rabbits as tumor group. For Gd-DTPA-enhanced MRI, T1WI and T2WI were performed on 9 animals of each model, and T1WI was acquired 80 seconds after administration of Gd-DTPA. For USPIO-enhanced MRI, T1WI, T2WI and T2*WI were performed on another 9 animals of each model before and 24 hours after administration of USPIO. MRI images were analyzed and correlated with surgical specimens and pathological results. RESULTS: In the tumor group, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of Gd-DTPA-enhanced MRI were 62.5%, 91.3%, 88.2%, 70.0% and 76.6%, respectively. The corresponding values of USPIO-enhanced MRI were 95.0%, 90.9%, 90.5%, 95.2% and 92.9%, respectively. The sensitivity and accuracy of USPIO-enhanced MRI differ significantly from those of Gd-DTPA-enhanced MRI. In the hyperplasia group, the accuracy of Gd-DTPA-enhanced MRI was 74.2%, while 87.1% for USPIO-enhanced MRI. CONCLUSION: USPIO-enhanced MRI has higher accuracy in diagnosing metastatic lymph nodes than Gd-DTPA-enhanced MRI.

Calcitonin gene-related peptide dynamics in rat dorsal root ganglia and spinal cord following different sciatic nerve injuries.

We examined calcitonin gene-related peptide (CGRP) expression dynamics in the dorsal root ganglia (DRGs) and spinal cords of adult rats subjected to one of the following three types of unilateral sciatic nerve injury: crush (SNC), ligation (SNL), or transection combined with subsequent neurorrhaphy (SNT). Following SNC, CGRP immunoreactivity (IR) was increased in ipsilateral primary sensory neurons of L4-L5 DRGs, laminae I-II and spinal motoneurons; an area of CGRP-labeled fibers in ipsilateral laminae III-V was also increased in size following SNC. CGRP up-regulation exhibited a distinct temporospatial pattern and expression levels had returned to baseline levels by the end of the 28-day test period. Similar to SNC, SNT also resulted in an increase of CGRP-IR in these areas, though to a slightly lesser degree in the three latter areas. By contrast SNL, which is associated with complete blockade of axonal transport, induced a sustained decrease in CGRP-IR in primary sensory neurons of L4-L5 DRGs and superficial laminae (I-IIo), as well as in an ipsilateral area occupied by CGRP-labeled fibers. Interestingly, SNL did not affect CGRP-IR in spinal motoneurons, but did result in an accumulation of nerve growth factor (NGF) distal to ligature that was apparent as early as 1 day post-injury and persisted throughout the experimental period. These findings indicate that the nature of peripheral nerve injury has an impact on CGRP expression dynamics and that the response involves target tissues in vivo. Our results have important implications for elucidating the mechanisms of nerve regeneration.

Dynamic changes in Robo2 and Slit1 expression in adult rat dorsal root ganglion and sciatic nerve after peripheral and central axonal injury.

Robos are transmembrane receptors that mediate Slit signaling to repel growth cone outgrowth and neural migration in the developing central nervous system. Their distribution and function in the peripheral nervous system remains unclear. In the present study, we examined expression of Slit1 and Robo2 in adult rat dorsal root ganglion (DRG), spinal cord and sciatic nerve after peripheral nerve injury (axotomy). In control rats, Slit1 and Robo2 mRNA and protein were expressed at basic levels in the L5 and L6 DRGs. Sciatic transection resulted in a significant up-regulation of both Robo2 and Slit1 mRNA and protein (p<0.05 versus control). The peak of Slit1 and Robo2 expression occurred at days 7 and 14, respectively, and returned to control levels at days 28 and 21 post-axotomy, respectively. By contrast, injury to the central axons of the DRG by dorsal rhizotomy did not up-regulate Slit1 and Robo2 expression. Robo2 staining was stronger in small diameter neurons than in large diameter neurons in control DRG. Interestingly, post-axotomy, Robo2 immunostaining increased in the large diameter neurons and the number of Robo2 positive large diameter neurons increased significantly relative to controls. Non-neuronal cells surrounding the primary sensory neurons, including the satellite cells, were Slit1-positive, and Slit1 protein was expressed in the myelin sheath and non-neural cells in both intact and degenerating sciatic nerve axons. Sciatic nerve transection also led to an accumulation of Slit1 protein in peripheral region of the traumatic neuroma. In conclusion, we report an altered expression and redistribution of Robo2 and Slit1 in the DRG and sciatic nerve trunk after peripheral axotomy. Our results indicate that Slit1 and Robo2 likely play an important role in regeneration after peripheral nerve injury.