IL-6 Up-Regulates Expression of LIM-Domain Only Protein 4 in Psoriatic Keratinocytes through Activation of the MEK/ERK/NF-κB Pathway.

Psoriasis is a chronic inflammatory skin disease characterized by the activation of keratinocytes and the infiltration of immune cells. Overexpression of the transcription factor LIM-domain only protein 4 (LMO4) promoted by IL-23 has critical roles in regulating the proliferation and differentiation of psoriatic keratinocytes. IL-6, an autocrine cytokine in psoriatic epidermis, is a key mediator of IL-23/T helper 17-driven cutaneous inflammation. However, little is known about how IL-6 regulates the up-regulation of LMO4 expression in psoriatic lesions. In this study, human immortalized keratinocyte cells, clinical biopsy specimens, and an animal model of psoriasis induced by imiquimod cream were used to investigate the role of IL-6 in the regulation of keratinocyte proliferation and differentiation. Psoriatic epidermis showed abnormal expression of IL-6 and LMO4. IL-6 up-regulated the expression of LMO4 and promoted keratinocyte proliferation and differentiation. Furthermore, in vitro and in vivo studies showed that IL-6 up-regulates LMO4 expression by activating the mitogen-activated extracellular signal-regulated kinase (MEK)/extracellular signal-regulated kinase (ERK)/NF-κB signaling pathway. These results suggest that IL-6 can activate the NF-κB signaling pathway, up-regulate the expression of LMO4, lead to abnormal proliferation and differentiation of keratinocytes, and promote the occurrence and development of psoriasis.

Pro-inflammatory cytokine IL-6 regulates LMO4 expression in psoriatic keratinocytes via AKT/STAT3 pathway.

The transcription factor LIM-only protein 4 (LMO4) is overexpressed in the psoriatic epidermis and regulates keratinocyte proliferation and differentiation. High LMO4 expression levels are induced by interleukin-23 (IL-23) to activate the AKT/STAT3 signaling pathway. Interleukin-6 (IL-6) is mainly involved in regulating T cell functions and development in patients with psoriasis. However, whether LMO4 expression is regulated by IL-6 remains unclear. Therefore, the purpose of this study is to explore the role and molecular mechanisms of IL-6 in regulating LMO4 expression. The interleukin-6 (IL-6) levels in human plasma were determined using a chemiluminescence immunoassay system. A psoriasis-like mouse model was established using imiquimod induction. Epidermal keratinocytes (HaCaT) were cultured in defined keratinocyte-serum-free medium and stimulated by IL-6 alone or with inhibitors. The proteins of interest were detected using western blot analysis, immunofluorescence, and immunohistochemistry. The 5-ethynyl-2'-deoxyuridine assay was used to detect cell proliferation. The results revealed that IL-6 levels were markedly increased in the plasma of patients with psoriasis, compared to healthy control. The high expression of LMO4 was consistent with high levels of IL-6, p-AKT, and p-STAT3 in the lesions of both psoriasis patients and imiquimod-induced psoriasis-like mice. IL-6 activates the AKT/STAT3 signaling pathway, followed by LMO4 high-expression in HaCaT cells. IL-6 induces HaCaT proliferation and differentiation via AKT/STAT3 signaling pathway activation. We think that the high expression of LMO4 in psoriatic keratinocytes requires IL-6 to activate the AKT/STAT3 signaling pathway and leads to epidermal keratinocytes abnormal proliferation and differentiation.

GPR108 is required for gambogic acid inhibiting NF-κB signaling in cancer.

GPCRs are the most potential targets for drug discovery, however, their role in oncology is underappreciated and GPCR-based anti-cancer drug is not fully investigated. Herein, we identified GPR108, a GPCR protein described in innate immune system, is a potential therapeutic target of cancer. Depletion of GPR108 dramatically inhibited the survival of various cancers. Notably, TNFα activation of NF-κB was totally impaired after GPR108 knockout. We identified gambogic acid (GA), a natural prenylated xanthone, selectively targeting GPR108. Importantly, GA engaged with GPR108 and promoted its degradation, knockout of GPR108 remarkably blocked GA inhibition of NF-κB signaling. Furthermore, in vitro and in vivo assays demonstrated that GA was dependent on GPR108 to exert anti-cancer activity. Overall, our findings supported GPR108 as a promising therapeutic target of cancer, and provided a small molecule inhibitor GA directly and selectively targeting GPR108 for cancer therapy.

A novel four-gene signature predicts immunotherapy response of patients with different cancers.

BACKGROUND: Immune checkpoint blockade (ICB) therapy has demonstrated favorable clinical efficacy, particularly for advanced or difficult-to-treat cancer types. However, this therapy is ineffective for many patients displaying lack of immune response or resistance to ICB. This study aimed to establish a novel four-gene signature (CD8A, CD8B, TCF7, and LEF1) to provide a prognostic immunotherapy biomarker for different cancers. METHODS: Transcriptome profiles and clinical data were obtained from The Cancer Genome Atlas database. Multivariate Cox regression analysis was used to establish a four-gene signature. The R package estimate was used to obtain the immune score for every patient. RESULTS: Risk scores of the novel four-gene signature could effectively divided all patients into high- and low-risk groups, with distinct outcomes. The immune score calculated via the estimate package demonstrated that the four-gene signature was significantly associated with the immune infiltration level. Furthermore, the four-gene signature could predict the response to atezolizumab immunotherapy in patients with metastatic urothelial cancer. CONCLUSIONS: The novel four-gene signature developed in this study is a good prognostic biomarker, as it could identify many kinds of patients with cancer who are likely to respond to and benefit from immunotherapy.

Combined short- and long-axis method for internal jugular vein catheterization in premature newborns: A randomized controlled trial.

BACKGROUND: Rapid central venous catheterization is critical for the rescue and perioperative management of premature infants requiring surgery. Ultrasound-guided dynamic needle tip positioning (DNTP) has been widely used as a very effective technique, especially in paediatric vascular puncture and catheterization. However, for low-weight premature newborns, central vein catheterization still poses greater difficulties for paediatricians and paediatric anaesthesiologists. This prospective randomized control study aimed to evaluate the efficacy of combined short- and long-axis (CSLA) internal jugular vein catheterization for premature newborns in comparison with the DNTP technique. METHODS: A total of 90 premature newborns (gestational age < 37 weeks and < 28 days after birth) who were scheduled for surgery were included in this study. All enrolled premature newborns were randomly divided into two groups (n = 45): the CSLA group and the DNTP group. We compared the first-puncture success rate, total success rate, procedure time, number of needle passes, occurrence of complications and other outcome measures between the two groups. RESULTS: The two groups (n = 45 per group) were similar in sex, gestational age, weight, mean arterial blood pressure, and vein-related measurements of the internal jugular vein. Total success was achieved in 43 (95.6%) and 36 (80.0%) patients in the CSLA and DNTP groups respectively. Compared with the DNTP group, the CSLA group showed a significantly higher first-attempt success rate (71.1% vs 46.7%, χ2  = 5.5533, P = .0184) and significantly fewer needle passes (1.0[1.0-2.0] vs 2.0[1.0-3.0], χ2  = -2.6094, P = .0091). There was no significant difference between the groups in the procedure time (368[304-573] vs 478[324-79]s, Z = -1.7690, P = .0769). Complications occurred in both groups, but the incidence was significantly lower in the CSLA group than in the DNTP group (6.7% vs 22.2%, χ2  = 4.4056, P = .0358). CONCLUSIONS: Ultrasound-guided internal jugular vein catheterization by the CSLA method is effective and safe. The CSLA method may be superior to the DNTP technique in premature newborns.

The therapeutic effect of controlled reoxygenation on chronic hypoxia-associated brain injury.

Cardiopulmonary bypass (CPB) is the most general technique applied in congenital heart disease (CHD). However, standard CPB poses a specific pathologic condition for patients during surgery: exposure to reoxygenation. When surgery is performed on cyanotic infants, standard CPB is usually initiated at a high concentration of oxygen without consideration of cytotoxic effects. Controlled reoxygenation is defined as using normoxic CPB with a pump primed to the PO2 (oxygen tension in the blood), which is matched to the patient's preoperative saturation. The aim of this study was to determine whether controlled reoxygenation could avoid standard reoxygenation injury and also to clarify the molecular signaling pathways during hypoxia. We successfully reproduced the abnormal brain observed in mice with chronic hypoxia during early postnatal development - equivalent to the third trimester in human. Mice were treated with standard reoxygenation and controlled reoxygenation after hypoxia for 24 h. We then assessed the brain tissue of these mice. In standard reoxygenation-treated hypoxia mice, the caspase-3-dependent neuronal apoptosis was enhanced by increasing concentration of oxygen. Interestingly, controlled reoxygenation inhibited neuron and glial cell apoptosis through suppressing cleavage of caspase-3 and PARP. We also found that controlled reoxygenation suppressed LCN2 expression and inflammatory cytokine (including TNF-α, IL-6, and CXCL10) production, in which the JAK2/STAT3 signaling pathway might participate. In conclusion, our findings propose the novel therapeutic potential of controlled reoxygenation on CPB during CHD.

LMO4 Is a Disease-Provocative Transcription Coregulator Activated by IL-23 in Psoriatic Keratinocytes.

Psoriasis is an autoimmune disease characterized by abnormal differentiation and hyperproliferation of epidermal keratinocytes. LIM-domain only protein 4 (LMO4) is a transcription factor coregulator that promotes the assembly of multiprotein complexes to regulate mammary epithelium and keratinocyte differentiation and proliferation during embryogenesis. In this study, LMO4 has been found to be abundantly expressed in psoriatic epidermis. LMO4 expression is increased in human keratinocytes induced to differentiate by calcium ex vivo, and LMO4 overexpression induces spontaneous differentiation and growth acceleration of human keratinocytes in the absence of calcium. IL-23, a cytokine highly expressed in psoriatic skin lesions, induces differentiation and promotes proliferation of human keratinocytes. The IL-23-mediated effects are accompanied by an increase in LMO4 expression mediated by signal transducer and activator of transcription 3 through an IL-23/acutely transforming retrovirus AKT8 in rodent T-cell lymphoma/signal transducer and activator of transcription 3 pathway in keratinocytes. Knockdown of LMO4 effectively inhibits differentiation and growth of keratinocytes both ex vivo and in IL-23-injected ears of mice. LMO4 appears to mediate IL-23-related responses in psoriatic keratinocytes and is a potential therapeutic target in psoriasis.

`Folate-Conjugated Phosphorescent Silica Nanoparticles for Bioimaging and Cancer Cell Targeting.

Besides their superior photophysical properties, phosphorescent transition-metal complexes have some drawbacks for biological applications because of their poor solubility in aqueous solutions and toxicity. In order to avoid the biological environmental impacts on their optical function and solve the problems of water-solubility and toxicity in sensing and bioimaging, two biocompatible organic/inorganic hybrid phosphorescent nanoprobes (Ir-MSN) by using mesoporous silica nanoparticles (MSN) as carriers and an iridium(III) complex as signaling unit were prepared in the present study. In addition, folic acid (FA) was covalently attached to them to endow the particles with characteristic of tumor targeting. The photophysical properties, cell staining and tumor cell targeting functions of FA-ligated Ir-MSN were investigated. These results demonstrated that such a design strategy of phosphorescent nanoprobes is an effective way to develop excellent phosphorescent cellular probes for living cell applications.

Grhl3 induces human epithelial tumor cell migration and invasion via downregulation of E-cadherin.

Grainyhead genes are involved in wound healing and developmental neural tube closure. Metastasis is a multistep process during which cancer cells disseminate from the site of primary tumors and establish secondary tumors in distant organs. The adhesion protein E-cadherin plays an essential role in metastasis. In light of the high degree of similarity between the epithelial-mesenchymal transition (EMT) occurring in wound-healing processes and the EMT occurring during the acquisition of invasiveness in skin or breast cancer, we investigated the role of the Grainyhead genes in cancer invasion. Here, we show that there is an inverse relationship between Grainyhead-like 3 (Grhl3) and E-cadherin expression in some epithelial tumor cell lines. Overexpression of Grhl3 in the E-cadherin-positive epithelial tumor cell line, characterized by less invasiveness, generated a transcriptional blockage of the E-cadherin gene and promoted cell migration and cell invasion. Conversely, Grhl3 depletion inhibited cell migration and cell invasion and was associated with a gain of E-cadherin expression. To further explore the mechanism by which Grhl3 regulated E-cadherin expression, an E-cadherin promoter report analysis was performed and results showed that Grhl3 repressed E-cadherin gene expression by directly or indirectly binding to the E-boxes present in the proximal E-cadherin promoter. Taken together, our findings define a major role for Grhl3 in the induction of migration and invasion by the downregulation of E-cadherin in cancer cells.

NFκB up-regulation of glucose transporter 3 is essential for hyperactive mammalian target of rapamycin-induced aerobic glycolysis and tumor growth.

Accumulating evidence indicates that mammalian target of rapamycin (mTOR) exerts a crucial role in aerobic glycolysis and tumorigenesis, but the underlying mechanisms remain largely obscure. Results from Tsc1- or Tsc2-null mouse embryonic fibroblasts (MEFs) and human cancer cell lines consistently indicate that the expression of glucose transporter 3 (Glut3) is dramatically up-regulated by mTOR. The rapamycin-sensitive mTOR complex 1 (mTORC1), but not the rapamycin-insensitive mTOR complex 2 (mTORC2), was involved in the regulation of Glut3 expression. Moreover, mTORC1 enhances Glut3 expression through the activation of the IKK/NFκB pathway. Depletion of Glut3 led to the suppression of aerobic glycolysis, the inhibition of cell proliferation and colony formation, and the attenuation of the tumorigenic potential of the cells with aberrantly hyper-activated mTORC1 signaling in nude mice. We conclude that Glut3 is a downstream target of mTORC1, and it is critical for oncogenic mTORC1-mediated aerobic glycolysis and tumorigenesis. Hence Glut3 may be a potential target for therapy against cancers caused by the aberrantly activated mTORC1 signaling.

Therapeutic effect of the treatment for colorectal cancer with adenoviral vectors mediated estrogen receptor ß gene therapy combined with thermotherapy.

INTRODUCTION: Our preliminary study on repressing colorectal tumors by recombinant adenoviruses (Ads) delivering the human ERß gene (Ad-ERß) has achieved positive result. METHODS: In this study, hydrophobic fluorescent dyes ICG-Der-01 was entrapped into the N-succinyl-N'-octyl chitosan (SOC) micelles to form the near infrared absorbing dyes SOC-ICG-Der-01 and SOC-ICG-Der-01 mediated near infrared laser (SOC-ICG-Der-01/NIR) thermotherapy was combined with Ad-ERß gene therapy to regress colon cancer in vivo. RESULTS: Firstly, the antitumor efficacies of SOC-ICG-Der-01/NIR thermotherapy were investigated on S180 ascites tumor-bearing mice. Results indicated that, the average tumor volume of SOC-ICG-Der-01/NIR group was the smallest among the three treatment groups. Then, thermotherapy with SOC-ICG-Der-01/NIR combined with Ad-ERß gene therapy to treat HCT-116 colon cancer xenograft model was investigated. Further results demonstrated that, SOC-ICG-Der-01/NIR thermotherapy showed the significantly inhibitory efficiency compared with control group and Ad-ERß enhanced the therapeutic effect of SOC-ICG-Der-01/NIR. CONCLUSION: These findings demonstrated that combined administration of Ad-ERß with SOC-ICG-Der-01/NIR thermotherapy represents a promising colon cancer therapeutic strategy.

Inhibitory effects of ERß on proliferation, invasion, and tumor formation of MCF-7 breast cancer cells--prognostication for the use of ERß-selective therapy.

CONTEXT: Estrogen is well-known as an important factor in the physiological functions and pathological processes of breast. Estrogen receptor ß (ERß) is expressed in the majority of breast cancers at lower levels compared with the normal breast tissue. OBJECTIVE: The effect of ERß on the characteristics of breast tumor cells and its prognostication for the use of ERß-selective therapy were investigated here for the first time. MATERIALS AND METHODS: ERß was overexpressed in ERα positive MCF-7 breast cancer cells by gene transfection. The proliferation, motility, and xenografts growth of MCF-7 cells were investigated by MTT assays, wound-healing assay and animal study. RESULTS: Results demonstrated that ERß-GFP localized in both the cytoplasm and the nucleus in the presence of 17ß-estradiol (E2), with stronger fluorescence-signal intensity in the nucleus, 2.8-times higher than that in the cytoplasm. The ERß overexpressed MCF-7 cells resulted in a 38.7% decreased growth rate and motility in vitro. Furthermore, ERß overexpression enhanced the antiproliferative effects of phytoestrogen, antiestrogen, and histone deacetylase inhibitor. Exogenous ERß expression reduced tumor volume by 99% at 27 days postadministration, indicated that overexpression of ERß led to retardation of tumor formation and growth in immunodeficient mice. DISCUSSION AND CONCLUSION: This study provided a relatively new evidence to support that ERß is an important modulator of proliferation and motility of breast cancer cells, and implied for the first time a possibility for the use of novel ERß-selective therapies in breast cancer treatment.

The enhanced antiproliferative response to combined treatment of trichostatin A with raloxifene in MCF-7 breast cancer cells and its relevance to estrogen receptor ß expression.

Antiestrogen is one type of the endocrine therapeutic agents for estrogen receptor α (ERα)-positive breast cancer. Unfortunately, this treatment alone is insufficient. Here we reported a novel potential anticancer strategy by using histone deacetylase (HDAC) inhibitor to enhance the action of endocrine therapy in ERα-positive breast cancer cell. The well-described HDAC inhibitor, trichostatin A (TSA), and antiestrogen raloxifene were found to, respectively, inhibit E2-induced proliferation of MCF-7 breast cancer cell in a dose-responsive and time-dependent manner. TSA and raloxifene enhanced the antiproliferative activity of each other by promoting cell death via apoptosis and cell cycle arrest. Thus, they displayed better antiproliferative effects in combined treatment than that with either agent alone. The expression level of estrogen receptor ß (ERß) showed a marked increase after TSA or/and raloxifene treatment. Treatments with TSA or/and raloxifene resulting in the up-regulation of ERß are in accordance with the antiproliferative effects of the two agents. Furthermore, the over-expression of ERß by adenovirus delivery could inhibit the proliferation of MCF-7 tumor cells and drastically enhanced the antiproliferative effects of TSA and raloxifene. These results demonstrated that the interference of ERß on the antiproliferative effects of HDAC inhibitor and antiestrogen constitutes a promising approach for breast cancer treatment.

Estrogen receptor ß potentiates the antiproliferative effect of raloxifene and affects the cell migration and invasion in HCT-116 colon cancer cells.

BACKGROUND: Estrogen receptor ß (ERß) is the predominant ER in the colorectal epithelium, whose expression is greatly reduced in colorectal cancer compared with normal colon tissue. Recent in vitro studies suggested that ERß may suppress tumor growth. No research was reported whether ERß can be used as therapeutic agent for colon cancer. METHODS: In this study, ERß gene constructed into adenoviral (Ad) vectors was used to treat colon cancer HCT-116 cells alone or in combination with raloxifene. In vitro and in vivo studies were conducted to investigate the therapeutic effects of ERß and raloxifene in HCT-116 cells. RESULTS: Our results indicated that, although Ad-ERß alone had no effect on the proliferation of HCT-116 cells, the combination of Ad-ERß with raloxifene significantly inhibited the proliferation of HCT-116 cells. The apparently apoptotic induction effects may partly explain the cytotoxicity of the two agents. The results of the study of ERß on migration and invasion of HCT-116 cells demonstrated that overexpression of ERß significantly decreased cell migration and increased invasion of cells. The antitumor efficacies of ERß as well as raloxifene were further investigated on HCT-116 tumor bearing mice. Results demonstrated that both Ad-ERß and raloxifene individually inhibited tumor growth. The combination group showed the highest inhibitory efficiency compared with other three groups. CONCLUSION: These findings demonstrated that combined administration of Ad-ERß with raloxifene represents a promising colon cancer therapeutic strategy.

In vivo monitoring of organ-selective distribution of CdHgTe/SiO2 nanoparticles in mouse model.

CdHgTe/SiO(2) nanoparticles were prepared by SiO(2) capping on the surface of CdHgTe QDs. The characteristics, such as optical spectra, photostability, size and cell toxicity were investigated. The dynamic distribution of CdHgTe/SiO(2) nanoparticles was in vivo monitored by near infrared fluorescence imaging system. CdHgTe/SiO(2) nanoparticles acted as a novel fluorescence probe have a maximum fluorescence emission of 785 nm and high photo-stability. The hydrodynamic diameter of CdHgTe/SiO(2) nanoparticles could be adjusted to 122.3 nm. Compared to CdHgTe QDs, inhibitory effects of CdHgTe/SiO(2) nanoparticles on proliferation of HCT116 cells decreased to a certain extent. CdHgTe/SiO(2) nanoparticles had their specific dynamic distribution behavior, which provided new perspectives for bio-distribution of nanoparticles.