Abstract Background Familial Hypocalciuric Hypercalcemia (FHH) is a group of rare inherited diseases caused by loss-of-function mutations in the calcium-sensing receptor (CaSR) gene or related proteins. A significant link has not been discovered between germline inactivating CaSR mutations and parathyroid tumors in sporadic PHPT. However, it has been observed that most adenomatous and hyperplastic tumors have decreased CaSR expression. Case presentation A 59-year-old woman was referred to our clinic for evaluation of hypercalcemia despite having no active complaints. She underwent total thyroidectomy and parathyroidectomy due to parathyroid adenoma and multinodular goiter. Pathology revealed a parathyroid lipoadenoma. Postoperatively, calcium levels did not return to physiological ranges, raising suspicion of FHH. Genetic analysis identified compound heterozygosity for CASR c.665G > A (p.Gly222Glu) and CASR c.2027 C > G (p.Tyr676Arg) variants, confirming the diagnosis of FHH. Conclusion Co-existence of PHPT due to a parathyroid lipoadenoma and FHH is rare. Patients may still exhibit mild to moderate hypercalcemia following surgery, thus more invasive procedures should be avoided; failure of parathyroid exploration should not be assumed.

The mechanical state of pre-tumoral epithelia controls subsequent Drosophila tumor aggressiveness.

Mouse mammary glands comprise ductal trees, which are lined by epithelial cells and have one opening at the tip of each nipple. The epithelial cells play a major role in mammary gland function and are the origin of most mammary tumors. Introducing genes of interest into mouse mammary epithelial cells is a critical step in evaluating gene function in epithelial cells and generating mouse mammary tumor models. This goal can be accomplished through the intraductal injection of a viral vector carrying the genes of interest into the mouse mammary ductal tree. The injected virus subsequently infects mammary epithelial cells, bringing in the genes of interest. The viral vector can be lentiviral, retroviral, adenoviral, or adenovirus-associated viral (AAV). This study demonstrates how a gene of interest is delivered into mammary epithelial cells through mouse mammary intraductal injection of a viral vector. A lentivirus carrying GFP is used to show stable expression of a delivered gene, and a retrovirus carrying Erbb2 (HER2/Neu) is used to demonstrate oncogene-induced atypical hyperplastic lesions and mammary tumors.

The incidence of hyperparathyroidism has increased in the USA. The previous work from our institution detected environmental chemicals (EC) within hyperplastic parathyroid tumors. The National Health and Nutrition Examination Survey (NHANES) is a program designed to assess the health status of people in the USA and includes measurements of EC in serum. Our aim was to determine which EC are associated with elevated parathyroid hormone (PTH) and calcium levels within NHANES. NHANES was queried from 2003-2016 for our analysis with calcium. A separate subgroup was queried from 2003-2006 that included PTH levels. Subjects with elevated calcium, and elevated PTH and normal Vitamin D levels were identified. Wilcoxon rank sum tests were used to analyze levels of EC in those with elevated calcium, and those with elevated PTH in the subgroup. All EC with p < 0.05 were then included in separate multivariate models adjusting for serum vitamin D and creatinine for PTH and albumin for calcium. There were 51,395 subjects analyzed, and calcium was elevated in 2.1% (1080) of subjects. Our subgroup analysis analyzed 14,681 subjects, and PTH was elevated without deficient Vitamin D in 9.4% (1,377). Twenty-nine different polychlorinated biphenyls and the organochlorine pesticides hexachlorobenzene, transnonachlor, oxychlordane, and p,p'-dichlorodiphenyldichloroethylene (DDE) were found to be associated with elevated calcium and separately with elevated PTH (all p < 0.05). In NHANES, 33 ECs were found to be associated with elevated calcium as well as elevated PTH levels on our subgroup analysis. These chemicals may lead us toward a causal link between environmental factors and the development of hyperparathyroidism and should be the focus of future studies looking at chemical levels within specimens.

The tumor suppressor gene phosphatase and tensin homolog (PTEN) in PI3K/Akt/mTOR pathway is essential in inhibiting tumor growth and metastasis. However, whether the mutation of PTEN gene could induce tumorigenesis and impact the treatment of gastric cancer is still unclear. The purpose of the study was to investigate the combined treatment of gastric tumorigenesis using Rapamycin and Fluorouracil (5-Fu) through interfering with the Akt/mTOR pathway in a mouse model with PTEN conditional deletion. Three groups of mice were exposed for 5days to Rapamycin and 5-Fu separately and together. The gene expression of the Akt/mTOR pathway, the protein expression of caspase-3 and p-Akt, p-S6K and p-4EBP1, and the pathological changes in stomachs were analyzed. Our study demonstrates that the conditional PTEN deletion in the cells of glandular stomach induces hyperplastic gastric tumors in mice. The combined Rapamycin administration with 5-Fu resulted in better outcomes than their separate administration for the treatment of gastric cancer by inhibiting the mTOR signal pathway. Our study indicates that Rapamycin has a synergistic interaction with chemotherapeutic 5-Fu, and demonstrates a potential therapeutic combination treatment on glandular stomach tumor with PTEN functional absence or aberrantly activated Akt/mTOR pathway. It provides important insights into the inhibition of the Akt/mTOR pathway in gastric cancer clinical therapy.

Background: A cylindrical piezoelectric element and a specific multi-channel circular microelectromechanical systems (MEMS)-transducer array of ultrasonic system were used for ultrasonic energy generation and ablation. A relatively long time is required for the heat to be conducted to the target position. Ultrasound thermal therapy has great potential for treating deep hyperplastic tissues and tumors, such as breast cancer and liver tumors. Methods: Ultrasound ablation technology produces thermal energy by heating the surface of a target, and the heat gradually penetrates to the target’s interior. Beamforming was performed to observe energy distribution. A resonance method was used to generate ablation energy for verification. Energy was generated according to the coordinates of geometric graph positions to reach the ablation temperature. Results: The mean resonance frequency of Channels 1–8 was 2.5 MHz, and the cylindrical piezoelectric ultrasonic element of Channel A was 4.2546 Ω at 5.7946 MHz. High-intensity ultrasound has gradually been applied in clinical treatment. Widely adopted, ultrasonic hyperthermia involves the use of high-intensity ultrasound to heat tissues at 42–45 °C for 30–60 min. Conclusion: In the ultrasonic energy method, when the target position reaches a temperature that significantly reduces the cell viability (46.9 °C), protein surface modification occurs on the surface of the target.

The X-linked acrogigantism-associated gene gpr101 is a regulator of early embryonic development and growth in zebrafish

Environmental chemicals and metabolic disruption in primary and secondary human parathyroid tumors

Receptor tyrosine kinase signaling plays prominent roles in tumorigenesis, and activating oncogenic point mutations in the core pathway components Ras, Raf, or MEK are prevalent in many types of cancer. Intriguingly, however, analogous oncogenic mutations in the downstream effector kinase ERK have not been described or validated in vivo To determine if a point mutation could render ERK intrinsically active and oncogenic, we have assayed in Drosophila the effects of a mutation that confers constitutive activity upon a yeast ERK ortholog and has also been identified in a few human tumors. Our analyses indicate that a fly ERK ortholog harboring this mutation alone (RolledR80S), and more so in conjunction with the known sevenmaker mutation (RolledR80S+D334N), suppresses multiple phenotypes caused by loss of Ras-Raf-MEK pathway activity, consistent with an intrinsic activity that is independent of upstream signaling. Moreover, expression of RolledR80S and RolledR80S+D334N induces tissue overgrowth in an established Drosophila cancer model. Our findings thus demonstrate that activating mutations can bestow ERK with pro-proliferative, tumorigenic capabilities and suggest that Drosophila represents an effective experimental system for determining the oncogenicity of ERK mutants and their response to therapy.

Gastric polyps are commonly found on upper endoscopy. With the increasing use of proton pump inhibitors and decreasing incidence of Helicobacter pylori infection, fundic gland polyps are now the most common gastric polyps in Western countries. Most of the other polyps, such as hyperplastic polyps, gastric adenomas, and gastric neuroendocrine tumors (NETs), are strongly associated with the presence of chronic atrophic gastritis, commonly due to either H. pylori infection or autoimmune gastritis. Gastric NETs, previously termed carcinoids, are rare neoplasms that often present as polypoid lesions and can be subcategorized into three subtypes. The most common subtype, type 1, is associated with chronic atrophic gastritis and generally thought to have low malignant potential. Type 2 NETs behave similarly to type I NETs but are specifically associated with the Zollinger-Ellison syndrome. Type 3 NETs are sporadic and highly malignant. All gastric polypoid lesions require histopathologic examination for diagnosis. The key aspect to the management of gastric NETs and other gastric polyps is to determine the malignant potential of the lesion in question. This then informs whether the patient needs removal of additional polyps if multiple, whether the patient needs further endoscopic surveillance, or whether surgery is indicated. This review contains 5 figures, 5 tables, and 50 references. Key Words: atrophic gastritis, fundic gland polyp, gastric adenoma, gastric carcinoid, gastric neuroendocrine tumor, gastric polyp, hyperplastic polyp, intestinal metaplasia

Gastric polyps are commonly found on upper endoscopy. With the increasing use of proton pump inhibitors and decreasing incidence of Helicobacter pylori infection, fundic gland polyps are now the most common gastric polyps in Western countries. Most of the other polyps, such as hyperplastic polyps, gastric adenomas, and gastric neuroendocrine tumors (NETs), are strongly associated with the presence of chronic atrophic gastritis, commonly due to either H. pylori infection or autoimmune gastritis. Gastric NETs, previously termed carcinoids, are rare neoplasms that often present as polypoid lesions and can be subcategorized into three subtypes. The most common subtype, type 1, is associated with chronic atrophic gastritis and generally thought to have low malignant potential. Type 2 NETs behave similarly to type I NETs but are specifically associated with the Zollinger-Ellison syndrome. Type 3 NETs are sporadic and highly malignant. All gastric polypoid lesions require histopathologic examination for diagnosis. The key aspect to the management of gastric NETs and other gastric polyps is to determine the malignant potential of the lesion in question. This then informs whether the patient needs removal of additional polyps if multiple, whether the patient needs further endoscopic surveillance, or whether surgery is indicated. This review contains 5 figures, 5 tables, and 50 references. Key Words: atrophic gastritis, fundic gland polyp, gastric adenoma, gastric carcinoid, gastric neuroendocrine tumor, gastric polyp, hyperplastic polyp, intestinal metaplasia

Abstract Introduction: BET inhibitors have raised high expectations for cancer treatment given their anti-proliferative effect by inhibiting BRD4 controlled enhancer activity of highly transcribed genes such as MYC(N). However, current inhibitors also target BRD2 and BRD3 which are functionally nonredundant with BRD4. In neuroblastoma only MYCN amplified tumors respond well to these drugs. Methods: We performed an integrated bioinformatics approach to scrutinize BET family genes as well as further candidate epigenetic regulators as targets for novel therapies in neuroblastoma. Results: First we performed a time-resolved expression data analysis of week 1 and 2 hyperplastic lesions and tumors derived from the TH-MYCN transgenic mouse model and confirmed dynamic regulation during tumor development for established neuroblastoma oncogenes and tumor suppressor genes. Next, we filtered within the highest upregulated genes for Cancer Gene Census (CGC) genes and identified 21 upregulated CGC genes mainly involved in chromatin remodeling and DNA repair. Finally, after further selection based on expression in CCLE and survival in neuroblastoma patients, BRD3 was identified as the top-ranked candidate. BRD3 exhibits drastic upregulation during tumor formation. Elevated BRD3 expression is the highest expressed gene in neuroblastoma cell lines upon analysis of the CCLE panel and associated with very poor prognosis. To explore the nonredundant functions of BRD3 in relation to BRD4, we performed RNA-sequencing after stable knockdown of BRD3 in neuroblastoma cell lines and compared the downstream effects on the transcriptome as well as the impact on cell viability to knockdown of BRD4 and pharmacological treatment with BET-inhibitors (JQ1, OTX015). In addition, we dissected the BRD3 protein complex by means of label-free mass spectrometry analysis to gain further insights into the BRD3 specific functions in relation to control of gene transcription and putative interaction with transcription factors such as MYCN. Current efforts are ongoing to test cooperative interaction of BRD3 versus BRD4 in dbh-MYCN driven neuroblastoma formation in zebrafish as well as BRD3 and BRD4 ChIP-sequencing in neuroblastoma cells. Conclusion: We identified BRD3 as a candidate novel driver gene in neuroblastoma and will present differential transcriptional control and protein interactions of BRD3 versus BRD4. This study can open the way towards developing BRD3 specific inhibitors for neuroblastoma and other BRD3 overexpressing cancers such as T-ALL and small cell lung carcinoma. Citation Format: Kaat Durinck, Jolien Dewyn, Anneleen Beckers, Siebe Loontiens, Suzanne Vanhauwaert, Daniel Carter, Belamy Chueng, Glenn Marshall, Katleen Depreter, Frank Westermann, Frank Speleman. BRD3 as a specific vulnerable therapeutic target in neuroblastoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1527. doi:10.1158/1538-7445.AM2017-1527

Induction of Chromosome Instability by Activation of Yes-Associated Protein and Forkhead Box M1 in Liver Cancer

Abstract Currently there is no cure for a metastatic disease and it is therefore critical to target the early events that foster metastasis. It is now also recognized that a favorable microenvironment in the metastatic site, primed by the tumor, is crucial for metastasis. Our study is geared towards deciphering cellular and molecular mechanisms governing the metastatic niche that may lead to novel targeted anti-metastatic therapeutics. We utilize the multi-stage MMTV-PyMT breast cancer mouse model, which shares significant similarities with human breast cancer. By injecting a reporter metastatic cell line into hyperplasia-bearing mice, we were able to probe the susceptibility of the lung microenvironment to metastatic seeding. We demonstrate that early during mammary tumorigenesis, before metastasis has occurred, a metastatic niche is formed in the lung microenvironment. This niche is initiated in part by tumor-induced systemic pro-inflammatory factors and local extracellular matrix remodelers, as matrix metalloproteinases (MMPs). We show that the metastatic niche is associated with MMP9-expressing CD11b+Gr1+ and other lung stromal cells. MMP9, which is also expressed by tumor cells, appears as a pivotal player in the process and is therefore considered as a desirable therapeutic target. To examine the role of MMP9 activity in lung metastatic colonization, we utilized novel endogenous-like, function-specific antibodies (SDS3) that block the transiently-activated enzyme conformation of MMP9, which presumably contributes to disease progression. The therapeutic potential of SDS3 has been demonstrated in models of inflammatory bowel disease. We show that metastatic seeding within the lung microenvironment can be inhibited by SDS3, not only in experimental metastasis models but also when the lung microenvironment is primed by hyperplastic mammary tumors. Primary tumor burden was not changed with SDS3, suggesting that blocking active MMP9 is effective in preventing early metastasis rather than established tumors. To study the biodistribution and pharmacokinetics of SDS3, we utilized whole-body bioluminescence, intravital and ex-vivo live microscopy as well as flow cytometry. We show that SDS3 is retained in myeloid cells within the microenvironment of mammary tumors and lung metastatic foci. In situ zymography shows high MMP activity in premetastaic MMTV-PyMT lungs, which is reduced after SDS3. SDS3 also inhibits colony formation of cultured metastatic cells. Our results suggest that a metastatic niche is present in the lungs of hyperplastic mammary tumor-bearing mice and that it can be targeted by blocking MMP9 activity. Our study offers new insights into effectively blocking the in vivo activity of dysregulated MMPs as early anti-metastatic therapy of various cancers. Citation Format: Vicki Plaks, Jonathan Chou, Carrie Maynard, Nguyen H. Nguyen, Niwen Kong, Inna Solomonov, Dalit Talmi-Frank, Caroline Bonnans, Irit Sagi, Zena Werb. Targeting matrix metalloproteinases (MMP) for anti-metastatic therapy: Blocking active MMP9 abrogates metastatic niche formation and prevents metastatic seeding in a breast cancer model. [abstract]. In: Proceedings of the 106th Annual Meeting of the American Association for Cancer Research; 2015 Apr 18-22; Philadelphia, PA. Philadelphia (PA): AACR; Cancer Res 2015;75(15 Suppl):Abstract nr 4724. doi:10.1158/1538-7445.AM2015-4724

Neuroblastoma is the one of the most common extracranial childhood malignancies, accounting for ∼15% of tumor-associated deaths in children. It is generally considered that neuroblastoma originates from neural crest cells in the paravertebral sympathetic ganglia and the adrenal medulla. However, the mechanism by which neuroblastoma arises during sympathetic neurogenesis and the cellular mechanism that drives neuroblastoma development remains unclear. The present study investigated the cell components during neuroblastoma development in the tyrosine hydroxylase-v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (TH-MYCN) mouse model, a transgenic mouse model of human neuroblastoma. The present study demonstrates that paired-like homeobox 2b (Phox2B)+ neuronal progenitors are the major cellular population in hyperplastic lesions and primary tumors. In addition, Phox2B+ neuronal progenitors in hyperplastic lesions or primary tumors were observed to be in an actively proliferative and undifferentiated state. The current study also demonstrated that high expression levels of Phox2B promotes neuroblastoma cell proliferation and xenograft tumor growth. These findings indicate that the proliferation of undifferentiated Phox2B+ neuronal progenitors is a cellular mechanism that promotes neuroblastoma development and indicates that Phox2B is a critical regulator in neuroblastoma pathogenesis.

4-vinyl-2, 6-dimethoxyphenol (canolol) is an antioxidant phenolic compound extracted from crude canola oil. In current research, K19-C2mE transgenic mice, developing hyperplastic tumors spontaneously in the glandular stomach, were used to study the mechanisms involved in the anti-inflammation and anti-tumor effects of canolol. Tg mice receiving canolol diet had a reduced tumor incidence, to 41.2%, compared with Non-treatment Tg mice, 77.8% of which had gastric tumor (P=0.002). Besides that, the mean tumor diameter was decreased from 6.5mm to 4.5mm (P<0.001) after canolol administration. COX-2/PGE2 pathway is known to play pivotal role in inflammation-induced gastric tumorigenesis. The neutrophils and lymphocytes infiltration was suppressed significantly, and the mRNA levels of the proinflammatory cytokines COX-2, IL-1β and IL-12b were also downregulated in gastric mucosa. Additionally, immunohistochemical analysis showed that COX-2, EP2, Gαs and β-catenin, key factors involving in PGE2 signal transduction, were positive staining with higher H scores in Non-treatment Tg mice, while the expressions were suppressed significantly by 0.1% canolol (P<0.001). In addition, tumor-suppressor miR-7 was reactivated after canolol administration, and COX-2 was showed to be a functional target of miR-7 to suppress the tumor progression. In conclusion, canolol could inhibit the gastritis-related tumor initiation and progression, and the suppression effect was correlated with the blocking up of canonical COX-2/PGE2 signaling pathway and might be regulated by miR-7.

Canolol inhibits hyperplastic gastric tumors initiation and progression in K19-C2mE transgenic mic

Abstract Recent studies have identified the involvement of receptor tyrosine kinases (RTKs) and their downstream signaling pathways, including MAPK, PI3K/Akt/mTOR, and Stat3, as key regulatory factors of epidermal proliferative capacity during skin carcinogenesis. Through development and study of transgenic mice with overexpression of a constitutively active form of Stat3 (Stat3C) in the basal cell layer, previous studies in the laboratory discovered that constitutive Stat3 activation accelerates tumor progression in the two-stage skin carcinogenesis model, which utilizes the tumor initiator 7,12-dimethylbenz[α]anthracene (DMBA) and tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA). Twist1, a well-known inducer of EMT, has been shown to be regulated by Stat3 in a number of systems. Western blot analysis of epidermal protein lysates from Stat3C transgenic mice indicated a two-fold induction of Twist1 expression as compared to wild type mice. In addition, immunofluorescence staining analysis of tumors from wild type mice [papillomas and squamous cell carcinomas (SCCs)] and Stat3C mice (SCCs) showed nuclear co-localization of Twist1 and Stat3. In the present study, we investigated the activation and subsequent downstream signaling of Twist1 during TPA-induced epidermal hyperproliferation. Western blot analysis of epidermal lysates from mice treated topically with multiple applications of TPA showed an initial decrease (4-6 h) followed by a subsequent rebound of Twist1 protein at 18 hours post TPA treatment. This initial decrease and then rebound coincided with that of the cell cycle regulatory proteins cyclin D1, cyclin E1, E2F-1, and c-myc. Interestingly, basal levels of Twist1 were reduced in epidermis specific Stat3 deficient mice. Furthermore, Stat3 deficient mice treated with TPA failed to show a rebound in expression of Twist1 in the epidermis. Immunofluorescence experiments of TPA treated epidermis revealed that nuclear localization of Twist1 was restricted to the proliferative compartment. Collectively, these results, combined with other data to be presented, suggest that Stat3 regulates basal levels of Twist1. This protein is localized in the nucleus of cells found primarily in the proliferative compartment of hyperplastic epidermis and skin tumors. The current results further suggest that Twist1 may play a role in cell cycle progression in keratinocytes during tumor promotion. Research supported by CA76520. Citation Format: Jaya Srivastava, Everardo Macias, Dharanija Rao, Kaoru Kiguchi, John DiGiovanni. Investigations to evaluate the role of Twist1 during tumor promotion and epithelial carcinogenesis. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 1290. doi:10.1158/1538-7445.AM2013-1290

Heme oxygenase-1 (HMOX1) is a cytoprotective enzyme degrading heme to biliverdin, iron ions, and carbon monoxide, whose expression is induced in response to oxidative stress. Its overexpression has been suggested as a strategy improving survival of transplanted muscle precursors. Here we demonstrated that HMOX1 inhibits differentiation of myoblasts and modulates miRNA processing: downregulates Lin28 and DGCR8, lowers the total pool of cellular miRNAs, and specifically blocks induction of myomirs. Genetic or pharmacological activation of HMOX1 in C2C12 cells reduces the abundance of miR-1, miR-133a, miR-133b, and miR-206, which is accompanied by augmented production of SDF-1 and miR-146a, decreased expression of MyoD, myogenin, and myosin, and disturbed formation of myotubes. Similar relationships between HMOX1 and myomirs were demonstrated in murine primary satellite cells isolated from skeletal muscles of HMOX1(+/+), HMOX1(+/-), and HMOX1(-/-) mice or in human rhabdomyosarcoma cell lines. Inhibition of myogenic development is independent of antioxidative properties of HMOX1. Instead it is mediated by CO-dependent inhibition of c/EBPδ binding to myoD promoter, can be imitated by SDF-1, and partially reversed by enforced expression of miR-133b and miR-206. Control C2C12 myoblasts injected to gastrocnemius muscles of NOD-SCID mice contribute to formation of muscle fibers. In contrast, HMOX1 overexpressing C2C12 myoblasts form fast growing, hyperplastic tumors, infiltrating the surrounding tissues, and disseminating to the lungs. We evidenced for the first time that HMOX1 inhibits differentiation of myoblasts, affects the miRNA processing enzymes, and modulates the miRNA transcriptome. HMOX1 improves the survival of myoblasts, but concurrently through regulation of myomirs, may act similarly to oncogenes, increasing the risk of hyperplastic growth of myogenic precursors.

BackgroundHedgehog signalling has been implicated in prostate tumorigenesis in human subjects and mouse models, but its effects on transforming normal basal/stem cells toward malignant cancer stem cells remain poorly understood.MethodsWe produced pCX-shh-IG mice that overexpress Hedgehog protein persistently in adult prostates, allowing for elucidation of the mechanism during prostate cancer initiation and progression. Various markers were used to characterize and confirm the transformation of normal prostate basal/stem cells into malignant cancer stem cells under the influence of Hedgehog overexpression.ResultsThe pCX-shh-IG mice developed prostatic intraepithelial neoplasia (PIN) that led to invasive and metastatic prostate cancers within 90 days. The prostate cancer was initiated through activation of P63+ basal/stem cells along with simultaneous activation of Hedgehog signalling members, suggesting that P63+/Patch1+ and P63+/Smo+ cells may serve as cancer-initiating cells and progress into malignant prostate cancer stem cells (PCSCs). In the hyperplastic lesions and tumors, the progeny of PCSCs differentiated into cells of basal-intermediate and intermediate-luminal characteristics, whereas rare ChgA+ neuroendocrine differentiation was seen. Furthermore, in the metastatic loci within lymph nodes, kidneys, and lungs, the P63+ PCSCs formed prostate-like glandular structures, characteristic of the primitive structures during early prostate development. Besides, androgen receptor (AR) expression was detected heterogeneously during tumor progression. The existence of P63+/AR-, CK14+/AR- and CD44+/AR- progeny indicates direct procurement of AR- malignant cancer trait.ConclusionsThese data support a cancer stem cell scenario in which Hedgehog signalling plays important roles in transforming normal prostate basal/stem cells into PCSCs and in the progression of PCSCs into metastatic tumor cells.