SCFFBXW11 Complex Targets Interleukin-17 Receptor A for Ubiquitin-Proteasome-Mediated Degradation.

Interleukin-17 (IL-17) is a pro-inflammatory cytokine that participates in innate and adaptive immune responses and plays an important role in host defense, autoimmune diseases, tissue regeneration, metabolic regulation, and tumor progression. Post-translational modifications (PTMs) are crucial for protein function, stability, cellular localization, cellular transduction, and cell death. However, PTMs of IL-17 receptor A (IL-17RA) have not been investigated. Here, we show that human IL-17RA was targeted by F-box and WD repeat domain-containing 11 (FBXW11) for ubiquitination, followed by proteasome-mediated degradation. We used bioinformatics tools and biochemical techniques to determine that FBXW11 ubiquitinated IL-17RA through a lysine 27-linked polyubiquitin chain, targeting IL-17RA for proteasomal degradation. Domain 665-804 of IL-17RA was critical for interaction with FBXW11 and subsequent ubiquitination. Our study demonstrates that FBXW11 regulates IL-17 signaling pathways at the IL-17RA level.

Defect of IL17 Signaling, but Not Centrinone, Inhibits the Development of Psoriasis and Skin Papilloma in Mouse Models.

Patients with psoriasis tend to develop skin cancer, and the hyperproliferation of the epidermis is a histopathological hallmark of both psoriasis and cutaneous squamous cell carcinoma (SCC), indicating that they may share pathogenic mechanisms. Interleukin-17 (IL17) stimulates the proliferation of the epidermis, leading to psoriasis. Overexpression of Polo-like kinase 4 (PLK4), which controls centriole duplication, has been identified in SCC, which also shows the hyperproliferation of keratinocytes. To investigate the cooperation between IL17 signaling and centriole duplication in epidermal proliferation, we established psoriasis and skin papilloma models in wild type (WT), IL17 receptor A (T779A) knockin (Il17ra(T779A)-KI), and IL17 receptor C knockout (Il17rc-KO) mouse strains. Bioinformatics, Western blot, immunohistochemical staining, colony formation, and real-time PCR were used to determine the effect of IL17 signaling and centrinone on epithelial proliferation. In the psoriasis model, compared to WT and Il17ra(T779A)-KI, Il17rc-KO dramatically suppressed epidermal thickening. The proliferation of keratinocytes significantly decreased in this order from WT to Il17ra(T779A)-KI and Il17rc-KO mice. In the skin papilloma model, Il17ra(T779A)-KI significantly decreased tumor burden compared to the WT, while Il17rc-KO abolished papilloma development. However, centrinone, a selective inhibitor of PLK4, did not affect skin lesion formation in either model. Our data demonstrated that Il17ra(T779A)-KI and Il17rc-KO prevent the development of psoriasis and tumorigenesis in the skin, while the topical administration of centrinone does not have any effect.

Nonlinear Inverse Analysis for Predicting the Tensile Properties of Strain-Softening and Strain-Hardening UHPFRC.

The tensile stress-strain response is considered to be the most important and fundamental mechanical property of ultra-high-performance fiber-reinforced concrete (UHPFRC). Nevertheless, it is still a challenging matter for researchers to determine the tensile properties of UHPFRC. As a simpler alternative to the direct tensile test, bending tests are widely performed to characterize the tensile behavior of UHPFRC, but require further consideration and a sophisticated inverse analysis procedure. In order to efficiently predict the tensile properties of UHPFRC, a nonlinear inverse method based on notched three-point bending tests (3PBT) was proposed in this paper. A total of fifteen UHPFRC beams were fabricated and tested to evaluate the sensitivity of the predicted tensile behavior to variations in fiber volume fraction. A segmented stress-strain model was used, which is capable of describing the various tensile properties of UHPFRC, including strain softening and strain hardening. A more approximate formulation was adopted to simulate the load-deflection response of UHPFRC beam specimens. The closed-form analytical solutions were validated by tensile test results and existing methods in literature. Finally, parametric studies were also conducted to investigate the robustness of the proposed method. The load-deflection responses obtained from notched 3PBT could be easily converted into tensile properties with this inverse method.

Cyclic Behavior of Reinforced High Strain-Hardening UHPC under Axial Tension.

The cyclic tensile behavior of steel-reinforced high strain-hardening ultrahigh-performance concrete (HSHUHPC) was investigated in this paper. In the experimental program, 12 HSHUHPC specimens concentrically placed in a single steel reinforcement under cyclic uniaxial tension were tested, accompanied by acoustic emission (AE) source locating technology, and 4 identical specimens under monotonic uniaxial tension were tested as references. The experimental variables mainly include the loading pattern, the diameter of the embedded steel rebar, and the level of target strain at each cycle. The tensile responses of the steel-reinforced HSHUHPC specimens were evaluated using multiple performance measures, including the failure pattern, load-strain response, residual strain, stiffness degradation, and the tension-stiffening behavior. The test results showed that the enhanced bond strength due to the inclusion of steel fibers transformed the failure pattern of the steel-reinforced HSHUHPC into a single, localized macro-crack in conjunction with a sprinkling of narrow and closely spaced micro-cracks, which intensified the strain concentration in the embedded steel rebar. Besides, it was observed that the larger the diameter of the embedded steel rebar, the smaller the maximum accumulative tensile strain under cyclic tension, which indicated that the larger the diameter of the embedded steel rebar, the greater the contribution to the tensile stiffness of steel-reinforced HSHUHPC specimens in the elastic-plastic stage. In addition, it was found that a larger embedded steel rebar appeared to reduce the tension-stiffening effect (peak tensile strength) of the HSHUHPC. Moreover, the residual strain and the stiffness of the steel-reinforced HSHUHPC were reduced by increasing the number of cycles and finally tended toward stability. Nevertheless, different target strain rates in each cycle resulted in different eventual cumulative tensile strain rates; hence the rules about failure pattern, residual strain, and loading stiffness were divergent. Finally, the relationship between the accumulative tensile strain and the loading stiffness degradation ratio under cyclic tension was proposed and the tension-stiffening effect was analyzed.

CDK4/6 inhibitor palbociclib reduces inflammation in lupus-prone mice.

Lupus is an autoimmune inflammatory disease that affects multiple organs. Cyclin-dependent kinases (CDKs) have been associated with inflammation. The objective of this study was to explore the effects of palbociclib (a CDK4/6 inhibitor) on inflammation in a lupus-prone MRL-lpr mouse model. Twenty mice (10 females and 10 males) were randomized into control group (n=10, treated with vehicle) and treatment group (n=10, treated with 3 cycles of palbociclib at a dose of 120 mg/kg/day for 2 weeks on and 10 weeks off, through oral gavage). Animals were euthanized after the third cycle of treatment. We found that facial skin lesions developed later and smaller in the female mice treated with palbociclib than the female mice in the control group. The lymph node number was less and the lymph node weight was lighter in the female treatment group compared to the female control group. The inflammatory lesions in the kidneys of male mice treated with palbociclib were significantly reduced compared to the male mice in the control group. Our findings suggest that palbociclib treatment reduces inflammation in lupus-prone mice in a gender-specific manner, targeting the facial skin and lymph nodes in the female mice and the kidneys in the male mice.

HOTAIR and androgen receptor synergistically increase GLI2 transcription to promote tumor angiogenesis and cancer stemness in renal cell carcinoma.

Tumor angiogenesis is a major characteristic of renal cell carcinoma (RCC). Herein, we report a novel mechanism of how lncRNA and androgen receptor (AR) drive the Hedgehog pathway to promote tumor angiogenesis in RCC. We found that the high expression of lncRNA HOTAIR in RCC is associated with poor prognosis. Moreover, HOTAIR and AR form a feedback loop to promote the expression of each other. Interestingly, we also found that in RCC, HOTAIR is associated with the Hedgehog pathway, especially GLI2, via bioinformatics analysis. Furthermore, HOTAIR promotes GLI2 expression in the presence of AR. Mechanistically, HOTAIR interacts with AR and they cooperatively bind to GLI2 promoter and increase its transcription activity. We further confirmed how HOTAIR-AR axis regulates GLI2 expression by analyzing its function in RCC cells and found that HOTAIR and AR synergistically enhanced the expression of GLI2 downstream genes, such as VEGFA, PDGFA, and cancer stem cell transcription factors, and promoted tumor angiogenesis and cancer stemness in RCC cells both in vitro and in tumor xenografts. Overall, these findings suggest that HOTAIR and GLI2 could be novel therapeutic targets against RCC.

CD82 Suppresses ADAM17-Dependent E-Cadherin Cleavage and Cell Migration in Prostate Cancer.

CD82 acts as a tumor suppressor in a series of steps in malignant progression. Here, we identified a novel function of CD82 on posttranslational regulating E-cadherin in prostate cancer. In our study, the declined expression of CD82 was verified in prostate cancer tissues and cell lines compared with normal tissue and cell lines. Functionally, CD82 inhibited cell migration and E-cadherin cleavage from the cell membrane in prostate cancer cell. Further study proved that a disintegrin and metalloproteinase ADAM17 as an executor of E-cadherin cleavage mediated the inhibitory regulation of CD82 in E-cadherin shedding in prostate cancer. Specifically, CD82 interacted with ADAM17 and inhibited its metalloprotease activity, which led to the descent of E-cadherin shedding. These results show a nuanced but important role of CD82 in nontranscriptional regulation of E-cadherin, which may help to understand the intricate regulation of dysfunctional adhesion molecule in cancer progression.

Intra-arterial injection to create bone metastasis of prostate cancer in mice.

Human prostate cancer often metastasizes to the bone, but the mechanisms are not quite clear. The difficulties in studying the biology of bone metastasis are due to lack of animal models with high frequency of bone metastases. In the present study, we tested two intra-arterial injection methods, i.e., intra-caudal artery injection and intra-femoral artery injection. Mouse prostate cancer cell line MPC3-luc was injected into C57BL/6J male mice via intra-caudal artery injection (n = 8) or intra-femoral artery injection (n = 11). We found one mouse developed metastatic tumors in both hind limbs and the tail after intra-caudal artery injection. Two mice developed metastatic tumors in the hind limb after intra-femoral artery injection. The metastatic tumors were detected by bioluminescent imaging and X-ray, and confirmed by histological examination. Our study finds that intra-arterial (either caudal or femoral artery) injection may be a useful model in studying prostate cancer bone metastasis, however, the injection technique is difficult.

Genome and transcriptome profiling of FBXW family in human prostate cancer.

F-box and WD repeat domain containing (FBXW) family of E3 ligases has 10 members that ubiquitinate substrate proteins for proteasome-mediated degradation. Publicly archived datasets from The Cancer Genome Atlas (TCGA), Prostate Cancer Transcriptome Atlas (PCTA), and cBioPortal were analyzed for mRNA expression and genetic alterations of 10 FBXW genes. We found that FBXW7 mRNA expression was significantly decreased in primary prostate cancers compared to normal prostate tissues, whereas mRNA expression of FBXW8-10 was significantly increased in primary prostate cancers compared to normal prostate tissues. FBXW7 mRNA expression was also significantly decreased in breast invasive carcinoma, glioblastoma multiforme, head and neck squamous cell carcinoma, lung squamous cell carcinoma, and uterine corpus endometrial carcinoma. In contrast, FBXW7 mRNA expression was significantly increased in cholangiocarcinoma, colon adenocarcinoma, kidney renal clear cell carcinoma, kidney renal papillary cell carcinoma, liver hepatocellular carcinoma, lung adenocarcinoma, pheochromocytoma and paraganglioma, and thyroid carcinoma. Compared to normal tissues, FBXW5 mRNA expression was significantly increased in breast invasive carcinoma, cholangiocarcinoma, kidney chromophobe, kidney renal clear cell carcinoma, liver hepatocellular carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, prostate adenocarcinoma, thyroid carcinoma, and uterine corpus endometrial carcinoma, whereas FBXW5 mRNA expression was only significantly decreased in colon adenocarcinoma. There were not any significant differences in gene copy number gains, losses, or gene simple somatic mutations between primary prostate cancers and normal prostate tissues. The mRNA expression levels of FBXW5, 7, 8, 9, and 12 were significantly higher in metastatic castration-resistant prostate cancers (mCRPCs) than primary prostate cancers, whereas mRNA expression levels of FBXW1 and 4 were significantly lower in mCRPCs than primary prostate cancers. All 10 FBXW genes had significantly more overall gene alterations including gene amplifications in mCRPCs than primary prostate cancers. FBXW5 and 7 had significantly more gene deep deletions in mCRPCs than primary prostate cancers and FBXW7 had significantly more gene missense mutations in mCRPCs than primary prostate cancers. Our findings suggest that different FBXW genes have differential mRNA expression in prostate cancer and other cancer types and their gene amplifications are significantly more in mCRPCs than primary prostate cancers. FBXW7 mRNA expression is consistently decreased in primary prostate cancers compared to normal prostate tissues.

The effects of exercise on insulin, glucose, IGF-axis and CRP in cancer survivors: Meta-analysis and meta-regression of randomised controlled trials.

BACKGROUND: The purpose of this study was to investigate the relationship between physical activity and biological mediators of cancer recurrence and survival. METHODS: We conducted a comprehensive literature search of PubMed, ScienceDirect and Web of Science for randomised controlled trials examining the association between physical activity and C-reactive protein (CRP), glucose, insulin, insulin resistance and insulin growth factor-one (IGF-1) up to December 2017. Standardised mean difference (SMD) scores were calculated, and meta-regression was performed. RESULTS: The meta-analysis indicated that survivors randomised to physical activity conditions experienced greater improvements in Insulin (SMD = -0.59; 95% CI, -1.05 to -0.14), CRP (SMD = -0.52; 95% CI, -0.87 to -0.17), insulin resistance (SMD = -0.20; 95% CI, -0.41 to -0.003) and glucose (SMD = -0.19; 95% CI, -0.35 to -0.02) than survivors randomised to control conditions. The meta-regression showed that study duration was positively, albeit marginally related (p = .056) to change in CRP levels among survivors in the physical activity conditions. Furthermore, higher baseline insulin levels in the physical activity conditions were associated with improving insulin levels throughout the intervention (p = .007). CONCLUSIONS: Promoting physical activity throughout the survivorship continuum is an effective intervention strategy for improving levels of insulin, glucose control, insulin resistance and CRP among cancer survivors.

Th17 cells promote tumor growth in an immunocompetent orthotopic mouse model of prostate cancer.

Interleukin-17 (IL-17) has been demonstrated to promote development of a variety of cancers including prostate cancer in genetically modified mouse models. IL-17 is the main product secreted by T helper 17 (Th17) cells. A recent study has shown that Th17 cells and related genes are upregulated in human prostate cancers. However, there is no direct experimental evidence to demonstrate Th17's role in prostate cancer. In the present study, we co-implanted mouse prostate cancer MPC3-luc cells with Th17-polarized mouse splenocytes in the prostate of immunocompetent C57BL/6J male mice. We found that Th17-polarized splenocytes promoted orthotopic allograft prostate tumor growth compared to the control splenocytes. The numbers of IL-17-positive lymphocytes and macrophages were higher in the prostate tumors grown from co-implantation of MPC3-luc cells and Th17-polarized splenocytes, compared to the prostate tumors grown from co-implantation of MPC3-luc cells and control splenocytes. Our findings provide the first direct experimental evidence that Th17 cells may promote prostate cancer growth.

Silibinin attenuates TGF­ß1­induced migration and invasion via EMT suppression and is associated with COX­2 downregulation in bladder transitional cell carcinoma.

Transforming growth factor (TGF)­ß1 is highly expressed in bladder transitional cell carcinoma (TCC) and is positively associated with tumor grade. TGF­ß1 signaling promotes cell metastasis by inducing epithelial­mesenchymal transition (EMT), however, the underlying mechanisms are not fully understood. Our previous study demonstrated the anti­metastatic effects of silibinin, a natural flavonoid derived from milk thistle, against TCC. The present study investigated the effects of silibinin on TGF­ß1­induced EMT in TCC, focusing on the role of prostaglandin­endoperoxide synthase 2 (COX­2). Cell migration was determined by a wound healing assay and Transwell migration assay, and cell invasion was investigated using a Transwell invasion assay. Cell morphology was observed using an inverted microscope. Cell viability was evaluated by an MTT and cell counting assays. EMT markers were detected by reverse transcription­quantitative polymerase chain reaction and western blotting. Specific small interfering RNA was used to knockdown COX­2 gene expression. TGF­ß1 promoted cell migration and invasion, induced EMT and upregulated the expression of COX­2. COX­2 knockdown attenuated TGF­ß1­induced EMT, indicating that COX­2 upregulation was essential for TGF­ß1­induced EMT. Silibinin attenuated TGF­ß1­induced migration and invasion by inhibiting EMT, and was associated with COX­2 downregulation. TGF­ß1­induced COX­2 upregulation, which was inhibited by silibinin. In addition, TGF­ß1­induced EMT was further inhibited when silibinin treatment was combined with COX­2­knockdown. The results suggested that silibinin may be a potential future treatment for metastatic TCC.

Synthesis of magnetic molecularly imprinted polymers with excellent biocompatibility for the selective separation and inhibition of testosterone in prostate cancer cells.

PURPOSE: Androgen plays an important role in the progression of prostate cancer. In the present study, novel magnetic molecularly imprinted polymers (MMIPs) with good biocompatibility were produced for the selective separation and inhibition of testosterone in prostate cancer cells. MATERIALS AND METHODS: MMIPs were prepared by using magnetic nanospheres, gelatin, and testosterone as the supporting materials, functional monomer, and the template molecule, respectively. The characterization of the resultant products was investigated by transmission electron microscopy, X-ray diffraction, and vibrating sample magnetometry. To test whether MMIPs can remove testosterone in biologic samples, human LNCaP (androgen-dependent) and C4-2 (androgen-independent) prostate cancer cells were selected as cell models. The translocation of androgen receptor (AR) was detected by immunofluorescence assay, and the expression of PSA mRNA was detected by real-time quantitative polymerase chain reaction analysis. Cell flow cytometry analysis was performed to detect cell cycle arrest. RESULTS: The synthesized nanomaterials (MMIPs) possessed high crystallinity, satisfactory superparamagnetic properties, and uniform imprinted shell, and exhibited high adsorption capacity, fast kinetics, and high selectivity for testosterone. Moreover, the obtained imprinted nanomaterials could selectively enrich and detect testosterone in the LNCaP cell samples as a solid-phase extractant coupled with high-performance liquid chromatography. In addition, the MMIPs could freely enter prostate cancer cells and suppress the translocation of AR into the cell nucleus. We further found that MMIPs inhibited upregulation of AR downstream target genes in LNCaP and C4-2 cells; also, MMIPs inhibited cell growth and induced obvious cell cycle arrest in androgen-dependent LNCaP cells, but had no obvious effect on androgen-independent C4-2 cells. CONCLUSION: Our results indicate that the obtained imprinted nanomaterials can specifically and effectively bind testosterone and recover it from prostate cancer cells. Moreover, the MMIPs can freely enter prostate cancer cells and block the activation of testosterone-AR pathway. Thus, the MMIPs may be a new option for antiandrogen therapy in prostate cancer.

Metformin potentiates the anticancer activities of gemcitabine and cisplatin against cholangiocarcinoma cells in vitro and in vivo.

Metformin, an oral biguanide drug used to treat type 2 diabetes, has displayed anticancer activities in several types of cancer cells. The combination of gemcitabine and cisplatin is the standard chemotherapy regimen for cholangiocarcinoma, but its benefit is limited. The present study aimed to investigate whether metformin could enhance the activities of gemcitabine and cisplatin against cholangiocarcinoma, and the underlying mechanisms. Metformin inhibited the proliferation of human cholangiocarcinoma RBE and HCCC-9810 cells and induced cell cycle arrest at the G0/G1 phase by increasing the activation of AMP-activated protein kinase (AMPK) pathways. Metformin upregulated the expression of p21Waf1 and p27kip1, and downregulated the expression of cyclin D1, a key protein required for cell cycle progression. The combination of gemcitabine and cisplatin inhibited the proliferation and induced the apoptosis of human cholangiocarcinoma cells by inducing the phosphorylation of AMPK, downregulating cyclin D1, and activating caspase-3. Administration of metformin enhanced the efficacy of gemcitabine and cisplatin to suppress the growth of cholangiocarcinoma tumors established in experimental models by inhibiting cell proliferation and inducing cell apoptosis through their effects on AMPK, cyclin D1 and caspase-3. Given that metformin has been used to treat type 2 diabetes patients for over half a century due to its superior safety profile, the results presented here indicate that metformin may be a potent agent for enhancing the efficacy of gemcitabine and cisplatin in the treatment of cholangiocarcinoma.