Jeong Keun Ahn

Antiviral effect of Curcuma longa Linn extract against hepatitis B virus replication

ETHNOPHARMACOLOGICAL RELEVANCE A medicinal herb Curcuma longa Linn has been used for treating various liver diseases caused by hepatitis B virus (HBV) in Asia. AIM OF THE STUDY The study was performed in order to investigate the antiviral activity of Curcuma longa Linn against HBV replication in liver cells. MATERIALS AND METHODS Aqueous extract of Curcuma longa Linn (CLL) was prepared and used to analyze its antiviral activity against HBV replication in HepG 2.2.15 cells, which contain HBV genomes. The inhibitory effect of CLL on HBV replication was examined by testing the levels of secreted HBV surface antigens (HBsAg), HBV DNAs, and HBV RNAs in HepG 2.2.15 cells using ELISA, Southern blot, and Northern blot analyses. Cytotoxic activities of CLL extract on various liver cells were analyzed by MTT assay. To dissect the inhibitory mechanism of CLL extract on HBV replication, the levels of p53 protein and p53 mRNAs were analyzed by Western blot and RT-PCR in HepG 2.2.15 cells. The repression of CLL extract on HBV transcription was analyzed by RT-PCR and CAT assay. RESULTS CLL extract repressed the secretion of HBsAg from HepG 2.2.15 cells. CLL extract also suppressed the production of HBV particles and the level of intracellular HBV RNAs in HepG 2.2.15 cells, suggesting that CLL extract inhibits HBV replication. We found that the anti-HBV activity of CLL extract is mediated through enhancing the cellular accumulation of p53 protein by transactivating the transcription of p53 gene as well as increasing the stability of p53 protein. It turned out that CLL extract repressed the transcription of HBx gene by suppressing HBV enhancer I and X promoter through p53 protein. In addition, CLL extract did not have any cytotoxic effects on liver cells. CONCLUSION These data showed that CLL extract represses HBV replication through enhancing the level of p53 protein. CLL extract can be used as a safe and specific drug for patients with liver diseases caused by HBV infection.

Hepatitis B virus X protein induces apoptosis by enhancing translocation of Bax to mitochondria

Hepatitis B virus X protein (HBx) is essential for viral replication and plays an important role in viral pathogenesis. HBx transactivates many viral and cellular genes and participates in cellular signal transduction pathways, proliferation, and apoptosis. In the present study, we report that HBx induces apoptosis by enhancing the translocation of Bax to mitochondria, followed by inducing the loss of mitochondrial membrane potential and release of cytochrome C. In addition, Bcl‐2, inhibitor of Bax, rescues the disruption of mitochondrial membrane potential and DNA fragmentation induced by serum starvation in HepG2‐X cells expressing HBx. We also found that HBx binds directly to Bax and interferes with the interaction between Bax and 14‐3‐3ε to enhance the translocation of Bax to mitochondria. Taken together, our data suggest that HBx induces apoptosis by interacting with Bax and enhancing its translocation to mitochondria.

Superparamagnetic γ-Fe2O3 nanoparticles as an easily recoverable catalyst for the chemical recycling of PET

There have been numerous studies to develop catalysts for the chemical recycling of poly(ethylene terephthalate) (PET) via glycolysis. However, in the field of PET glycolysis, only a few have attempted to recover and reuse the catalysts. This research utilized easily recoverable superparamagnetic γ-Fe2O3 nanoparticles as a reusable catalyst for PET glycolysis. γ-Fe2O3 nanoparticles were produced by calcining Fe3O4 nanoparticles prepared by the co-precipitation method. The produced γ-Fe2O3 nanoparticles had an average size of 10.5 ± 1.4 nm, and a very high surface area reaching 147 m2 g−1. Its superparamagnetic property was also confirmed. Glycolysis reactions were carried out, and the γ-Fe2O3 catalysts were recovered after the reactions by simple magnetic decantation. The use of magnetic iron oxide allowed the easy recovery of the catalyst from the glycolysis products. At 300 °C and a 0.05 catalyst/PET weight ratio, the maximum bis(2-hydroxyethlyl) terephthalate (BHET) monomer yield reached more than 90% in 60 min. At 255 °C and a 0.10 catalyst/PET weight ratio, the BHET yield reached more than 80% in 80 min. The catalyst was reused 10 times, giving almost the same BHET yield each time.

Aqueous extract of Tribulus terrestris Linn induces cell growth arrest and apoptosis by down-regulating NF-κB signaling in liver cancer cells

ETHNOPHARMACOLOGICAL RELEVANCE A medicinal herb Tribulus terrestris Linn has been used to treat various diseases including hepatocellular carcinoma. The aim of the present study was to investigate the anticancer activity of Tribulus terrestris Linn (TT) in liver cancer cells. MATERIALS AND METHODS The antitumor activity of aqueous TT extract was analyzed by testing the cytotoxicity and the effect on clonogenecity in HepG2 cells. Apoptosis and cell cycle arrest induced by TT were dissected by flow cytometry and its inhibitory effect on NF-κB activity was determined by analyzing the expression levels of NF-κB/IκB subunit proteins. The suppression of NF-κB-regulated gene expression by TT was assessed by RT-PCR. RESULTS TT extract repressed clonogenecity and proliferation, induced apoptosis, and enhanced accumulation in the G0/G1 phase of liver cancer cells. It also turned out that TT extract inhibited NF-κB-dependent reporter gene expression and NF-κB subunit p50 expression, while it enhanced the cellular level of IκBα by inhibiting the phosphorylation and degradation of IκBα. In addition, IKK activity was inhibited in a dose-dependent manner. Furthermore, TT extract suppressed the transcription of genes associated with cell cycle regulation, anti-apoptosis, and invasion. CONCLUSION These data showed that TT extract blocks proliferation and induces apoptosis in human liver cancer cells through the inhibition of NF-κB signaling. Aqueous TT extract can be used as an anticancer drug for hepatocellular carcinoma patients.

Herpes simplex virus type 1 ICP27 induces apoptotic cell death by increasing intracellular reactive oxygen species

HSV-1 ICP27, an immediate early protein of herpes simplex virus type 1, is involved in viral replication, transcriptional activation, RNA stability, apoptosis, and reactivation from viral latency. The reactivation from viral latency is closely related to apoptosis and oxidative stress. To understand the effect of ICP27 upon apoptosis, we tested for cell death of ICP27-expressing cells (3-3) in the presence of hydrogen peroxide. Under oxidative stress, 3-3 cells were sensitive to death, displaying typical apoptosis features such as oligonucleosomal DNA fragmentation, chromatin condensation, and G0/G1 accumulation. To dissect the sensitizing mechanism of ICP27 in apoptosis, we analyzed the level of intracellular reactive oxygen species (ROS) in 3-3 cells. We found that 3-3 cells exhibited increased ROS levels compared to Vero cells devoid of ICP27. We also observed that the increased levels of intracellular ROS in 3-3 cells were caused by disturbances in antioxidant enzymes. In 3-3 cells, the elevated ROS increased the AP-1 activity, subsequently the expression of Bax increased, and the expression of Bcl2 was reduced. In addition, 3-3 cells were sensitive to various apoptotic agents. Taken together, these results indicate that HSV-1 ICP27 sensitizes the cells to apoptosis by elevating the intracellular levels of ROS.

HSV-1 ICP27 induces apoptosis by promoting Bax translocation to mitochondria through interacting with 14-3-3θ

The subcellular localization of Bax plays a crucial role during apoptosis. In response to apoptotic stimuli, Bax translocates from the cytoplasm to the mitochondria, where it promotes the release of cytochrome c to the cytoplasm. In cells infected with HSV-1, apoptosis is triggered or blocked by diverse mechanisms. In this study, we demonstrate how HSV-1 ICP27 induces apoptosis and modulates mitochondrial membrane potential in HEK 293T cells. We found that ICP27 interacts with 14-3-3θ which sequesters Bax to the cytoplasm. In addition, ICP27 promotes the translocation of Bax to the mitochondria by inhibiting the interaction between 14-3-3θ and Bax. Our findings may provide a novel apoptotic regulatory pathway induced by ICP27 during HSV-1 infection.

VBP1 represses cancer metastasis by enhancing HIF‐1α degradation induced by pVHL

von Hippel‐Lindau‐binding protein 1 (VBP1) physically interacts with pVHL, an E3‐ubiquitin ligase, which degrades HIF‐1α in an oxygen‐dependent manner. HIF‐1 is a key regulator of adaptive responses to a lack of oxygen that controls glucose metabolism, angiogenesis, proliferation, invasion, and metastasis. However, the role of VBP1 in pVHL‐mediated degradation of HIF‐1α is not yet known. In this study, we show that VBP1 enhances the stability of pVHL and facilitates pVHL‐mediated ubiquitination of HIF‐1α. Furthermore, VBP1 suppresses HIF‐1α‐induced epithelial‐mesenchymal transition in vitro and tumor metastasis in vivo. These findings suggest that VBP1 is a bona fide tumor suppressor protein associated with HIF‐1α regulation.

SIRT2 reduces actin polymerization and cell migration through deacetylation and degradation of HSP90

SIRT2, a member of the class III histone deacetylase family, has been identified as a tumor suppressor, which is associated with various cellular processes including metabolism and proliferation. However, the effects of SIRT2 on cancer cell migration caused by cytoskeletal rearrangement remain uncertain. Here we show that SIRT2 inhibits cell motility by suppressing actin polymerization. SIRT2 regulates actin dynamics through HSP90 destabilization and subsequent repression of LIM kinase (LIMK) 1/cofilin pathway. SIRT2 directly interacts with HSP90 and regulates its acetylation and ubiquitination. In addition, the deacetylase activity of SIRT2 is required for the regulation of actin polymerization and the ubiquitin-mediated proteasomal degradation of HSP90 induced by SIRT2.

Effects of drop size and measuring condition on static contact angle measurement on a superhydrophobic surface with goniometric technique

It is not a simple task to measure a contact angle of a water drop on a superhydrophobic surface with sessile drop method, because a roll-off angle is very low. Usually contact angle of a water drop on a superhydrophobic surface is measured by fixing a drop with intentional defects on the surface or a needle. We examined the effects of drop size and measuring condition such as the use of a needle or defects on the static contact angle measurement on superhydrophobic surface. Results showed that the contact angles on a superhydrophobic surface remain almost constant within intrinsic measurement errors unless there is a wetting transition during the measurement. We expect that this study will provide a deeper understanding on the nature of the contact angle and convenient measurement of the contact angle on the superhydrophobic surface.