Pharmacological blockade of cholesterol trafficking by cepharanthine in endothelial cells suppresses angiogenesis and tumor growth

Lyu, Junfang,Yang, Eun Ju,Head, Sarah A.,Ai, Nana,Zhang, Baoyuan,Wu, Changjie,Li, Ruo-Jing,Liu, Yifan,Yang, Chen,Dang, Yongjun,Kwon, Ho Jeong,Ge, Wei,Liu, Jun O.,Shim, Joong Sup Elsevier 2017 Cancer letters Vol.409 No.-

<P><B>Abstract</B></P> <P>Cholesterol is an important modulator of membrane protein function and signaling in endothelial cells, thus making it an emerging target for anti-angiogenic agents. In this study, we employed a phenotypic screen that detects intracellular cholesterol distribution in endothelial cells (HUVEC) and identified 13 existing drugs as cholesterol trafficking inhibitors. Cepharanthine, an approved drug for anti-inflammatory and cancer management use, was amongst the candidates, which was selected for in-depth mechanistic studies to link cholesterol trafficking and angiogenesis. Cepharanthine inhibited the endolysosomal trafficking of free-cholesterol and low-density lipoprotein in HUVEC by binding to Niemann-Pick disease, type C1 (NPC1) protein and increasing the lysosomal pH. The blockade of cholesterol trafficking led to a cholesterol-dependent dissociation of mTOR from the lysosomes and inhibition of its downstream signaling. Cepharanthine inhibited angiogenesis in HUVEC and in zebrafish in a cholesterol-dependent manner. Furthermore, cepharanthine suppressed tumor growth in vivo by inhibiting angiogenesis and it enhanced the antitumor activity of the standard chemotherapy cisplatin in lung and breast cancer xenografts in mice. Altogether, these results strongly support the idea that cholesterol trafficking is a viable drug target for anti-angiogenesis and that the inhibitors identified among existing drugs, such as cepharanthine, could be potential anti-angiogenic and antitumor agents.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A phenotypic screen identified 13 existing drugs, including cepharanthine, as cholesterol trafficking inhibitors. </LI> <LI> Cepharanthine inhibited lysosomal cholesterol trafficking by binding to NPC1 protein and increasing the lysosomal pH. </LI> <LI> The blockade of cholesterol trafficking led to a cholesterol-dependent dissociation of mTOR from the lysosomes. </LI> <LI> Cepharanthine inhibited angiogenesis in HUVEC and in zebrafish in a cholesterol-dependent manner. </LI> <LI> Cepharanthine treatment enhanced the antitumor activity of cisplatin in lung and breast cancer xenografts in mice. </LI> </UL> </P>

Oxygen tank for synergistic hypoxia relief to enhance mitochondria-targeted photodynamic therapy

Xianghui Li,Haoran Wang,Zhiyan Li,Dandan Li,Xiaofeng Lu,Shichao Ai,Yuxiang Dong,Song Liu,Jinhui Wu,Wenxian Guan 한국생체재료학회 2022 생체재료학회지 Vol.26 No.4

Background: Mitochondria play an essential role in cellular redox homeostasis maintenance and meanwhile serve as an important target for organelle targeted therapy. Photodynamic therapy (PDT) is a promising strategy for organelle targeted therapy with noninvasive nature and highly spatiotemporal selectivity. However, the efficacy of PDT is not fully achieved due to tumor hypoxia. Moreover, aerobic respiration constantly consumes oxygen and leads to a lower oxygen concentration in mitochondria, which continuously limited the therapeutic effects of PDT. The lack of organelle specific oxygen delivery method remains a main challenge. Methods: Herein, an Oxygen Tank is developed to achieve the organelle targeted synergistic hypoxia reversal strategy, which not only act as an oxygen storage tank to open sources and reduce expenditure, but also coated with red blood cell membrane like the tank with stealth coating. Within the oxygen tank, a mitochondrion targeted photosensitizer (IR780) and a mitochondria respiration inhibitor (atovaquone, ATO) are co-loaded in the RBC membrane (RBCm) coated perfluorocarbon (PFC) liposome core. Results: Inside these bio-mimic nanoparticles, ATO effectively inhibits mitochondrial respiration and economized endogenous oxygen consumption, while PFC supplied high-capacity exogenous oxygen. These Oxygen modulators reverse the hypoxia status in vitro and in vivo, and exhibited a superior anti-tumor activity by mitochondria targeted PDT via IR780. Ultimately, the anti-tumor effects towards gastric cancer and colon cancer are elicited in vivo. Conclusions: This oxygen tank both increases exogeneous oxygen supply and decreases endogenous oxygen consumption, may offer a novel solution for organelle targeted therapies.

Gene Silencing of β-catenin by RNAi Inhibits Proliferation of Human Esophageal Cancer Cells by Inducing G0/G1 Cell Cycle Arrest

Wang, Jin-Sheng,Ji, Ai-Fang,Wan, Hong-Jun,Lu, Ya-Li,Yang, Jian-Zhou,Ma, Li-Li,Wang, Yong-Jin,Wei, Wu Asian Pacific Journal of Cancer Prevention 2012 Asian Pacific journal of cancer prevention Vol.13 No.6

Objectives: The aim of the present study was to explore mechanisms underlying the effects of down-regulating ${\beta}$-catenin expression on esophageal carcinoma (EC) cells. Methods: Cell cycle distribution and apoptosis were determined using flow cytometry and annexin V apoptosis assay, respectively. Transmission electron microscopy (TEM) was used to examine changes in ultrastructure, while expression of cyclin D1 protein and mRNA was detected by western blot and real-time PCR. Proliferating cell nuclear antigen (PCNA) and extracellular signal-regulated kinase (ERK) 1-2 were evaluated by Western blot analysis. PCNA labeling index (LI) was determined by immunocytochemistry. Results: Compared with pGen-3-con transfected and Eca-109 cells, the percentage of G0/G1-phase pGen-3-CTNNB1 transfected cells was obviously increased (P<0.05), with no significant difference among the three groups with regard to apoptosis (P>0.05). pGen-3-CTNNB1 transfected cells exhibited obvious decrease in cyclin D1 mRNA and protein expression (P<0.05) and the ultrastructure of Eca-109 cells underwent a significant change after being transfected with pGen-3-CTNNB1, suggesting that down-regulating ${\beta}$-catenin expression can promote the differentiation and maturation. The expression of PCNA and the ERKI/2 phosphorylation state were also down-regulated in pGen-3-CTNNB1 transfected cells (P<0.05). At the same time, the PCNA labeling index was decreased accordingly (P<0.05). Conclusion: Inhibition of EC Eca-109 cellproliferation by down-regulating ${\beta}$-catenin expression could improve cell ultrastructure by mediating blockade in G0/G1 through inhibiting cyclin D1, PCNA and the MAPK pathway (p-ERK1/2).

Real-time modeling prediction for excavation behavior

Li-FengNi,Ai-QunLi,Fu-YiLiu,HonoreYin,J.R.Wu 국제구조공학회 2003 Structural Engineering and Mechanics, An Int'l Jou Vol.16 No.6

Exogenous hydrogen sulfide improves chilling tolerance by regulating hydrogen peroxide production in cucumber seedlings

Wu Guoxiu,Li Shengli,Dong Yaoxin,Bi Huangai,Ai Xizhen 한국원예학회 2022 Horticulture, Environment, and Biotechnology Vol.63 No.5

Hydrogen sulfi de (H 2 S) and hydrogen peroxide (H 2 O 2 ) play signifi cant roles as signaling molecules in multiple physiological activities. However, the mechanism and relationship between H 2 S and H 2 O 2 in regulating chilling tolerance remain unclear. This study found that chilling stress induced both endogenous H 2 S and H 2 O 2 systems in cucumber seedlings. Exogenous NaHS (an H 2 S donor) treatment notably strengthened the mRNA abundance of respiratory burst oxidase homolog ( RBOH ), contributing to endogenous H 2 O 2 accumulation. However, H 2 O 2 had little eff ect on mRNA abundances and activities of L-/D-cysteine desulfhydrase (L/DCD), and endogenous H 2 S levels. NaHS and H 2 O 2 increased chilling tolerance in cucumber seedlings, as shown by decreased electrolyte leakage (EL), chilling injury indexes, reactive oxygen species (ROS), and malonaldehyde (MDA) content, and improved activities and expression levels of antioxidant enzymes as well as glutathion (GSH) and ascorbic acid (AsA) contents. Additionally, the photosynthetic rates (Pn), actual photochemical effi ciency (φPSII), maximum photochemical effi ciency (Fv/Fm), and mRNA abundances and activities of Calvin cycle enzymes (Rubisco, SBPase, FBA, and TK) were induced by NaHS and H 2 O 2 . Interestingly, the H 2 S-induced chilling tolerance was suppressed by dimethylthiourea (DMTU, an H 2 O 2 scavenger). The results illustrated that H 2 S repressed the negative eff ects of chilling stress by improving photosynthesis and alleviating oxidative damage, and H 2 O 2 may be a downstream signaling molecule in this process.

Protection Against Helicobacter pylori Infection by a Trivalent Fusion Vaccine Based on a Fragment of Urease B-UreB414

Li Wang,Xiao-Fei Liu,Shi Yun,Xiao-Peng Yuan,Xu-Hu Mao,Chao Wu,Wei-Jun Zhang,Kai-Yun Liu,Gang Guo,Dong-Shui Lu,Wen-De Tong,Ai-Dong Wen,Quan-Ming Zou 한국미생물학회 2010 The journal of microbiology Vol.48 No.2

A multivalent fusion vaccine is a promising option for protection against Helicobacter pylori infection. In this study, UreB414 was identified as an antigenic fragment of urease B subunit (UreB) and it induced an antibody inhibiting urease activity. Immunization with UreB414 partially protected mice from H. pylori infection. Furthermore, a trivalent fusion vaccine was constructed by genetically linking heat shock protein A (HspA), H. pylori adhesin A (HpaA), and UreB414, resulting in recombinant HspA-HpaA-UreB414 (rHHU). Its protective effect against H. pylori infection was tested in BALB/c mice. Oral administration of rHHU significantly protected mice from H. pylori infection, which was associated with H. pylori-specific antibody production and Th1/Th2-type immune responses. The results show that a trivalent fusion vaccine efficiently combats H. pylori infection, and that an antigenic fragment of the protein can be used instead of the whole protein to construct a multivalent vaccine.

The seismic responses of girder bridges with novel sliding lead rubber bearings

Wu, Yi-feng,Li, Ai-qun,Wang, Hao Techno-Press 2021 Structural monitoring and maintenance Vol.8 No.3

Based on the commonly used lead rubber bearing (LRB) and sliding rubber bearing (SRB), a novel sliding lead rubber bearing (SLRB) is introduced. The mechanical properties of the three types of bearings were investigated by experiment. After that, a simply supported girder bridge with a 1/4 scale ratio was designed and fabricated, and the dynamic characteristics and seismic response of the bridge equipped with the above three types of bearings were studied. Results show that the girder's acceleration response has been effectively reduced by setting bearings only for relatively high earthquake intensity. Compared with LRB and SRB, SLRB works with more compositive seismic isolation effect. The "slide" action of the telflon-stainless-steel interface in SLRB can significantly reduce the acceleration response of girder, while the relative displacement between the pier and girder for this novel bearing is not increased due to the occurrence of collision in the bearing.

The compression-shear properties of small-size seismic isolation rubber bearings for bridges

Wu, Yi-feng,Wang, Hao,Sha, Ben,Zhang, Rui-jun,Li, Ai-qun Techno-Press 2018 Structural monitoring and maintenance Vol.5 No.1

Taking three types of bridge bearings with diameter being 100 mm as examples, the theoretical analysis, the experimental research as well as the numerical simulation of these bearings is conducted. Since the normal compression and shear machines cannot be applied to the small-size bearings, an improved equipment to test the properties of these bearings is proposed and fabricated. Besides, the simulation of the bearings is conducted based on the explicit finite element software ANSYS/LS-DYNA, and some parameters of the bearings are modified in the finite element model to reduce the computation cost effectively. Results show that all the research methods are capable of revealing the fundamental properties of the small-size bearings, and a combined use of these methods can better catch both the integral properties and the inner detailed mechanical behaviors of the bearings.

Promoted Growth of Maize by the Phosphate Solubilizing Bacteria Isolated from North-east China

Wu, Hai-Yan,Wang, Li-Chun,Gao, Xing-Ai,Jin, Rong-De,Fan, Zuo-Wei,Kim, Kil-Yong,Zhao, Lan-Po Korean Society of Soil Science and Fertilizer 2011 한국토양비료학회지 Vol.44 No.1

A strain of phosphate solubilizing bacterium was isolated from rhizosphere and identified as Burkholderia sp. by 16S-rRNA gene sequence analyses. The bacterium was found to release gluconic acid and the solubilization of hydroxyapatite in the liquid medium by a significant drop in pH to 3.7 from an initial pH 7.0. The soluble-P concentration continuously increased during the incubation periods and the total amount of soluble P released in culture filtrate was detected at 990 mg $L^{-1}$ after 10 days of inoculation. Most promoted maize growth was found in the standard NPK (240-120-120 kg $ha^{-1}$) soil inoculation with Burkholderia sp. (Twenty milliliters/plant, 106 CFU) and also in the absence of Burkholderia sp. inoculation, the soil amended with only 2/3 levels of P gave significant higher plant yield compared to 1/3 levels of P or without P supplementation.

Global Finite-time Stabilization for a Class of High-order Nonlinear Systems with Multiple Unknown Control Directions

Jing Li,Jian Wu,Xiguo Yuan,Xiaobo Li,Liefu Ai 제어·로봇·시스템학회 2017 International Journal of Control, Automation, and Vol.15 No.1

This paper considers the problem of the global finite-time stabilization for the high-order nonlinear systemswith unknown control directions. Due to the uncertainty of control directions, the paper analyzes all possibleconditions of the directions. The Lyapunov-based logic switching rule ensures that we can find the correct controldirections. The adaptive switching controller with a switching parameter which is to be tuned online guarantees thatthe derivative of Lyapunov function is less than a negative definite function and the closed-loop system is globallyfinite-time stable. The effectiveness of the proposed method is illustrated by an example.