Opposing roles of RUBCN isoforms in autophagy and memory B cell generation.

RUBCN (also known as Rubicon) was originally identified as a negative regulator of autophagy, a process by which cells degrade and recycle damaged components or organelles and that requires the activity of the class III PI3K VPS34 and the mTORC1 protein complex. Here, we characterized the role of a shorter isoform, RUBCN100, as an autophagy-promoting factor in B cells. RUBCN100 was translated from alternative translation initiation sites and lacked the RUN domain of the longer, previously characterized RUBCN130 isoform. Specific deficiency of RUBCN130 in B cells enhanced autophagy, which promoted memory B cell generation. In contrast to RUBCN130, which is localized in late endosomes and lysosomes and suppresses the enzymatic activity of VPS34, an effect thought to mediated by its RUN domain, RUBCN100 was preferentially located in early endosomes and enhanced VPS34 activity, presumably because of the absence of the RUN domain. Furthermore, RUBCN100, but not RUBCN130, enhanced autophagy and suppressed mTORC1 activation. Our findings reveal that the opposing roles of two RUBCN isoforms are critical for autophagy regulation and memory B cell differentiation.

The Extracts of Astragalus membranaceus Inhibit Melanogenesis through the ERK Signaling Pathway.

Melanin is a normal production protecting skin from environment-causing damage. Plants produce some agents in response to their environment. These agents could be applied in cosmetic production. Some Chinese herbals have immunomodulatory activities and modulate the symptoms of several diseases. Melanogenesis represents a complex group of conditions that are thought to be mediated through a complex network of regulatory processes. Previously, some studies found that the extracts of Astragalus membranaceus (PG2) regulated immunity and supported hematopoiesis. Herein, we want to determine the molecular mechanisms by which PG2 inhibits melanogenesis in B16F10 melanoma cells. The cellular melanin contents and expression of melanogenesis-related protein, including microphthalmia associated transcription factor (MITF) and tyrosinase were significantly reduced after PG2 treatment. Moreover, PG2 increased phosphorylation of ERK, without affecting phosphorylation of p38. These results suggested that PG2 as a new target in reducing hyperpigmentation through the ERK signal pathway. PG2 has potential for cosmetic usage in the future.

The inhibition of indoleamine 2, 3-dioxygenase 1 by connexin 43.

Upregulation of connexin 43 (Cx43) showed potential in enhancing immune surveillance that was suppressed in the tumor microenvironment. The expression of indoleamine 2, 3-dioxygenase (IDO) is one of the crucial factors contributing to tumor immune tolerance by depletion of tryptophan and IDO-mediated tryptophan metabolites. Here, we aim to investigate the role of Cx43 in IDO production in murine tumor by using Cx43 inducers. Resveratrol (trans-3, 5, 4 '-trihydroxystilbene) is a natural plant-derived polyphenol possessing positive effect against cancer. Salmonella enterica serovar choleraesuis (S.C.) was proved to target and inhibit tumor growth. Both of them regulated Cx43 expression in tumor cells and led to either chemosensitizing or immune-activating. In this study, the correlation between Cx43 and IDO were determined by the treatment of resveratrol and S.C. Our data showed an increase in Cx43 while IDO protein and IDO-mediated inhibited effects on T cell decreased after tumor cells are given with resveratrol and S.C. TREATMENTS: All of which could be inhibited once the expression of Cx43 was blocked. Cx43 involved in IDO regulation might be useful in developing IDO-targeted cancer immune therapy.

Salmonella overcomes tumor immune tolerance by inhibition of tumor indoleamine 2, 3-dioxygenase 1 expression.

Over the past decades, Salmonella has been proven capable of inhibiting tumor growth. It can specifically target tumors and due to its facultative anaerobic property, can be more penetrative than other drug therapies. However, the molecular mechanism by which Salmonella inhibits tumor growth is still incompletely known. The antitumor therapeutic effect mediated by Salmonella is associated with an inflammatory immune response at the tumor site and a T cell-dependent immune response. Many tumors have been proven to have a high expression of indoleamine 2, 3-dioxygenase 1 (IDO), which is a rate-limiting enzyme that catalyzes tryptophan to kynurenine, thus causing immune tolerance within the tumor microenvironment. With decreased expression of IDO, increased immune response can be observed, which might be helpful when developing cancer immunotherapy. The expression of IDO was decreased after tumor cells were infected with Salmonella. In addition, Western blot analysis showed that the expression levels of phospho-protein kinase B (P-AKT), phospho-mammalian targets of rapamycin (P-mTOR), and phospho-p70 ribosomal s6 kinase (P-p70s6K) in tumor cells were decreased after Salmonella infection. In conclusion, our results indicate that Salmonella inhibits IDO expression and plays a crucial role in anti-tumor therapy, which might be a promising strategy combined with other cancer treatments.

The treatment of mouse colorectal cancer by oral delivery tumor-targeting Salmonella.

Systemic administration of Salmonella to tumor-bearing mice leads to its preferential accumulation in tumor sites, the enhancement of host immunity, and the inhibition of tumor growth. However, the underlying mechanism for Salmonella-induced antitumor immune response via oral delivery remained uncertain. Herein, we used mouse colorectal cancer (CT26) as tumor model to study the therapeutic effects after oral delivery of Salmonella. When orally administered into tumor-bearing mice, Salmonella significantly accumulated in the tumor sites, inhibited tumor growth and extended the survival of mice. No obvious toxicity was observed during orally administered Salmonella by examining body weight and inflammatory cytokines. As indoleamine 2, 3-dioxygenase 1 (IDO) is a crucial mediator for tumor-mediated immune tolerance, we examined the expression of IDO. We demonstrated that Salmonella inhibited IDO expression in mouse cancer cells. Furthermore, immunohistochemical studies of the tumors revealed the infiltration of neutrophils and T cells in mice treated with Salmonella. In conclusion, our results indicate that Salmonella exerts its tumoricidal effects and stimulates T cell activities by inhibiting IDO expression. Oral delivery of Salmonella may, represent a potential strategy for the treatment of tumor.

The extract of Rhodobacter sphaeroides inhibits melanogenesis through the MEK/ERK signaling pathway.

Reducing hyperpigmentation has been a big issue for years. Even though pigmentation is a normal mechanism protecting skin from UV-causing DNA damage and oxidative stress, it is still an aesthetic problem for many people. Bacteria can produce some compounds in response to their environment. These compounds are widely used in cosmetic and pharmaceutical applications. Some probiotics have immunomodulatory activities and modulate the symptoms of several diseases. Previously, we found that the extracts of Rhodobacter sphaeroides (Lycogen™) inhibited nitric oxide production and inducible nitric-oxide synthase expression in activated macrophages. In this study, we sought to investigate an anti-melanogenic signaling pathway in α-melanocyte stimulating hormone (α-MSH)-treated B16F10 melanoma cells and zebrafish. Treatment with Lycogen™ inhibited the cellular melanin contents and expression of melanogenesis-related protein, including microphthalmia-associated transcription factor (MITF) and tyrosinase in B16F10 cells. Moreover, Lycogen™ reduced phosphorylation of MEK/ERK without affecting phosphorylation of p38. Meanwhile, Lycogen™ decreased zebrafish melanin expression in a dose-dependent manner. These findings establish Lycogen™ as a new target in melanogenesis and suggest a mechanism of action through the ERK signaling pathway. Our results suggested that Lycogen™ may have potential cosmetic usage in the future.

Salmonella enhance chemosensitivity in tumor through connexin 43 upregulation.

The use of preferentially replicating bacteria as oncolytic agents is one of the innovative approaches for the treatment of cancer. The capability of Salmonella to disperse within tumors and hence to delay tumor growth was augmented when combined with chemotherapy. This work is warranted to elucidate the underlying mechanism of antitumor effects by the combination therapy of Salmonella and cisplatin. The presence of functional gap junctions is highly relevant for the success of chemotherapy. Following Salmonella treatment, dose- and time-dependent upregulation of connexin 43 (Cx43) expressions were observed. Moreover, Salmonella significantly enhanced gap intercellular communication (GJIC), as revealed by the fluorescent dye scrape loading assay. To study the pathway underlying these Salmonella-induced effects, we found that Salmonella induced a significant increase in mitogen-activated protein kinases (MAPK) signaling pathways. The Salmonella-induced upregulation of Cx43 was prevented by treatment of cells with the phosphorylated p38 inhibitor, but not phosphorylated extracellular signal-regulated kinase (pERK) inhibitor or phosphorylated c-jun N terminal kinase (pJNK) inhibitor. Specific knockdown of Cx43 had an inhibitory effect on GJIC and resulted in a reduction of cell death after Salmonella and cisplatin treatment. Our results suggest that accumulation of Salmonella in tumor sites leads to increase Cx43 gap junction communication and enhances the combination of Salmonella and cisplatin therapeutic effects.

Tracking of mouse breast cancer stem-like cells with Salmonella.

Systemic administration of Salmonella to tumor-bearing mice leads to the preferential accumulation within tumor sites and retardation of tumor growth. The cancer stem-like cell (CSC) hypothesis suggests that CSCs are the root of cancer and induce metastasis and recurrence. The objective of this study was to examine if Salmonella could inhibit the growth of CSCs derived from mouse breast cancer. Systemically injected Salmonella preferentially accumulated within tumors for at least three weeks and the bacteria accumulated preferentially not only in subcutaneous but also in orthotopic tumors over livers and spleens at ratios ranging from 1000:1 to 10,000:1. Salmonella were capable of delaying tumor growth and enhancing survival in both subcutaneous and orthotopic tumor models. More strikingly, Salmonella acted to retard tumor growth and extensively prolong the survival time of the mice bearing CSC-induced tumors. Our results also found that Salmonella predominantly, although not exclusively, resided in the CSC regions of the tumor. These data suggest that Salmonella can inhibit the growth of breast cancer by targeting the CSC niche. In conclusion, Salmonella can be used for the management of breast cancer.

Resveratrol inhibits LPS-induced epithelial-mesenchymal transition in mouse melanoma model.

Epithelial to mesenchymal transition (EMT) has been linked to metastasis. Resveratrol exhibits potential antitumor activities; however, the inhibitory effects of resveratrol on the EMT of melanoma have not been demonstrated. Here, a new role for LPS in promoting EMT is described. LPS-induced EMT was identified by examining the markers of EMT. To assess the activation of NF-κB signal transduction pathway, we performed a reporter assay by using tumor cells transfected with the luciferase gene under the control of NF-κB response elements. The antitumor effects of resveratrol were evaluated in an experimental mouse metastasis tumor model. LPS increased N-cadherin and Snail expression and decreased zonula occludens-1 expression in a dose- and time-dependent manner. Meanwhile, LPS stimulated cell migration through activation of TLR4/NF-κB signal pathway. LPS-induced EMT is critical for inflammation-initiated metastasis. Nuclear translocation and transcriptional activity of p65 NF-κB, an important inducer of EMT, were inhibited by resveratrol. Resveratrol inhibited LPS-induced tumor migration and markers of EMT, significantly prolonged animal survival and reduced the tumor size. Thus, resveratrol plays an important role in the inhibition of LPS-induced EMT in mouse melanoma through the down-regulation of NF-κB activity. The data provide an insight into the mechanisms on the function of resveratrol during the processes of EMT.