Novel lignan glycosides 1a–1 h as analogues of cleistanthin A were designed and synthesized. Most of these compounds displayed significant cytotoxicities against four cancer cell lines. Compound 1e displayed better cytotoxicity than cleistanthin A with IC50 values from 1.0 nm to 8.3 nm. Further lysosome acidity assay disclosed that 1e could totally inhibit the activity of V‐ATPase at 60 nm.

Novel series of Farnesylthiosalicylic acid‐diamine/phenylpropenoic acid hybrids were designed and synthesized. Their in vitro growth inhibitory assays showed that most compounds displayed strong antiproliferation activity against seven cancer cells. Especially, the new hybrid 12f, by the conjugation of 10a with ferulic acid, could selectively suppress the proliferation of tumor cells and display significantly lower toxicities to normal cells than its intermediate 10a. Furthermore, 12f dose‐dependently induced SMMC‐7721 cell apoptosis. Additionally, our observations demonstrated that 12f inhibited both Ras‐related signaling and phosphorylated NF‐κB synergistically, which may be advantageous to the strong antitumor activities of 12f. Our findings suggest that these novel hybrids may hold a great promise as therapeutic agents for the intervention of human cancers.

A novel series of hybrids was designed and synthesized by combining key elements from farnesylthiosalicylic acid (FTS) and hydroxamic acid. Several 3,7,11-trimethyldodeca-2,6,10-trien-1-yl) thio)benzamide derivatives, particularly those with branched and linear aliphatic linkers between the hydroxamic zinc binding group (ZBG) and the benzamide core, not only displayed significant antitumor activities against six human cancer cells but also exhibited histone deacetylase (HDAC) inhibitory effects invitro. Among them, N-(4-(hydroxyamino)-4-oxobutyl)-2-(((2E,6E)-3,7,11-trimethyldodeca-2,6, 10-trien-1-yl)thio)benzamide (8d) was the most potent, with IC50 values of 4.9-7.6M; these activities are eight- to sixteen-fold more potent than FTS and comparable to that of suberoylanilide hydroxamic acid (SAHA). Derivative 8d induced cell cycle arrest in the G0/G1 phase, inhibited the acetylation of histone H3 and -tubulin, and blocked Ras-related signaling pathways in a dose-dependent manner. The improved tumor growth inhibition and cell-cycle arrest invitro might result from the dual inhibition. These findings suggest dual inhibitors of Ras-related signaling pathway and HDAC hold promise as therapeutic agents for the treatment of cancer.

E2F transcription factors regulate a wide range of biological processes, including cell cycle, apoptosis and DNA damage response. In the present study, we examined whether E2F2 is related to the poor prognosis of NSCLC and its role in progress of NSCLC. Firstly, we analyzed 86 NSCLC samples by immunohistochemistry and found that E2F2 expression was markedly increased in 62.8% (54/86) of all samples compared with the normal tissues. Further study showed that E2F2 expression was closely associated with clinical stage (P = 0.039) and tumor size (P = 0.045). Furthermore, Kaplan-Meier analysis indicated that high Bad expression was significantly correlated to overall survival (P = 0.045) but not disease-free survival (P = 0.288). In addition, our results showed that knockdown E2F2 expression could reduce cell viability and colony formation in NSCLC cells. The results in our study for the first time revealed that E2F2 act as an activator in tumor progress of NSCLC and could become a promising marker for the prognosis of patients with NSCLC.

Hypoxia is a characteristic of cancer and plays a key role in tumorigenesis, angiogenesis and resistance to cancer therapies. SiRNA treatment is effective against hypoxic tumors by gene silencing. However, siRNA delivery to the hypoxic regions of solid tumors still presents a challenge due to the distance from blood vessels and the increased presence of efflux transporters. Therefore, tumor therapies would be improved through the immediate development of an effective siRNA delivery system to hypoxic regions. To this end, we synthesized a system to deliver HIF-1α siRNA into hypoxic tumor cells. The system consists of a functional shell composed of 2-deoxyglucose (DG)-polyethylene glycol (PEG) connected with the compound of lipoic acid, lysine and 9-poly-d-arginine (LA-Lys-9R) by a hydrazone bond and a core of CdTe quantum dots (QDs). The molecular structure of DG-PEG-LA-Lys-9R was confirmed by liquid chromatography-mass spectrometry (LC-MS), nuclear magnetic resonance (NMR) spectroscopy, Fourier transform infrared spectroscopy (FTIR), and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The multifunctional CdTe QDs measured approximately 200 nm and showed excellent biocompatibility, perfect siRNA binding capability and enhanced hypoxic tumor targeting. Importantly, the system described here is pH-responsive with a hydrazone bond; therefore, it avoids GLUT1 receptor-mediated endocytic recycling, resulting in irreversible delivery of the siRNA. We used Western blots to confirm the superior gene silencing efficiency induced by the DG-PEG-LA-Lys-9R with hydrazone modified CdTe QDs. Here, we demonstrate high efficacy of the siRNA tumor delivery system using in vitro and in vivo experiments. In addition, these studies demonstrate that pH-responsive hybrid quantum dots show improved antitumor efficacy with decreased organ toxicity, indicating a promising siRNA delivery system for hypoxic cancer therapy.

Novel farnesylthiosalicylic acid (FTS) derivatives were synthesized by coupling with different substituted diamines. Their in vitro growth inhibitory activities against seven human cancer cell lines were evaluated. The results revealed that the synthetic farnesylthiosalicylamides displayed significant antitumor activities compared to the positive control FTS. Especially, compound 8f exhibited the strongest antitumor activities with IC50 values of 6.20-7.83 µM, which were one- to threefold less than those of sorafenib and six- to tenfold less than that of FTS against each cell line in vitro. Furthermore, 8f could inhibit the Ras-related signaling pathway and induce SMMC-7721 cell apoptosis superior to FTS in a dose-dependent manner. These data indicate that 8f may hold greater promise as therapeutic agent for the intervention of human cancers.

通过偶联法尼基硫代水杨酸(FTS)的羧基与羟基肉桂酸的酚羟基,设计合成了16个新型FTS/羟基肉桂酸偶联物(7a～7p),并对其进行了体外抗肿瘤活性研究.结果表明,大部分化合物对6种人肿瘤细胞具有较强的抗增殖活性,其中,化合物7b表现出最佳的肿瘤细胞增殖抑制活性,对所测肿瘤细胞的IC5o为5.51～9.25 μmol/L,优于FTS和索拉非尼的活性.并且,化合物7b可以选择性地抑制肿瘤细胞的生长,而对正常细胞损伤较小.此外,流式细胞分析显示化合物7b可以浓度依赖性地诱导SMMC-7721细胞凋亡.

Previous studies showed that prostacyclin inhibited fibrosis. However, both receptors of prostacyclin, prostacyclin receptor (IP) and peroxisome proliferator-activated receptor (PPAR), are abundant in cardiac fibroblasts. Here we investigated which receptor was vital in the anti-fibrosis effect of prostacyclin. In addition, the possible mechanism involved in protective effects of prostacyclin against cardiac fibrosis was also studied. We found that beraprost, a prostacyclin analogue, inhibited angiotensin II (Ang II)-induced neonatal rat cardiac fibroblast proliferation in a concentration-dependent and time-dependent manner. Beraprost also suppressed Ang II-induced collagen I mRNA expression and protein synthesis in cardiac fibroblasts. After IP expression was knocked down by siRNA, Ang II-induced proliferation and collagen I synthesis could no longer be rescued by beraprost. However, treating cells with different specific inhibitors of PPAR subtypes prior to beraprost and Ang II stimulation, all of the above attenuating effects of beraprost were still available. Moreover, beraprost significantly blocked transforming growth factor β (TGF β) expression as well as Smad2 phosphorylation and reduced Smad-DNA binding activity. Beraprost also increased phosphorylation of cAMP response element binding protein (CREB) at Ser133 in the nucleus. Co-immunoprecipitation analysis revealed that beraprost increased CREB but decreased Smad2 binding to CREB-binding protein (CBP) in nucleus. In conclusion, beraprost inhibits cardiac fibroblast proliferation by activating IP and suppressing TGF β-Smad signal pathway.

Hybrid 5f significantly inhibited both Ras-related signaling and phosphorylated NF-κB, which may synergistically contribute to its apoptosis induction and tumor growth inhibition in vitro and in vivo.

N-acetyl-L-cysteine (NAC) capped quantum dots (QDs) were synthesized by a hydrothermal method and coated with 2-amino-2-deoxy-D-glucose (DG), polyethylene glycol (PEG), and 9-D-arginine (9R). The optical properties, morphology and structure of 9R/DG-coated CdTe QDs were characterized by ultraviolet-visible spectrometry, fluorescence spectrum, Fourier transform infrared (FTIR), proton nuclear magnetic resonance (1H NMR), liquid chromatography-mass spectrometer (LC-MS), sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transmission electron micrographs (TEM). Furthermore, the biocompatibility, tumor targeted ability and transmembrane action of 9R/DG-coated CdTe QDs were studied. Results indicated that 9R/DG-coated CdTe QDs was constructed successfully by ligand exchange. The 9R/DG-coated CdTe QDs with the size of 8-10 nm had good dispersity and the absorbance and fluorescence peaks of CdTe QDs after modification were red shifted from 480 nm to 510 nm and 627 nm to 659 nm, respectively. In addition, the CdTe QDs modified by PEG, DG and 9R displayed good biocompatibility, high targeted ability to the cancer cells with glucose transporter type 1 (GLUT1) receptor high expression and obvious transmembrane ability.

We sought to investigate whether TSG suppressed the ICAM-1/VCAM-1 expression in dietary atherosclerotic rats and in Ox-LDL-induced U937 cells. For this purpose, 60 male Sprague–Dawley rats were randomly-and-equally divided into six groups. Atherosclerosis was induced by feeding rats a hyperlipidemic diet. TSG (120, 60 or 30 mg/kg/day) was administered by oral gavage. Simvastatin (2 mg/kg/day) was administered as positive control whereas physiological saline (0.9 % NaCl) served as untreated control. After 12 weeks, rats were euthanized by ethyl carbonate (1,200 mg/kg) and aortic wall samples were collected. Besides, U937 cells were stimulated for 48 h by Ox-LDL (80 μg/mL) with and without TSG (120, 60, 30 μg/L) or simvastatin (100 μg/L). ICAM-1/VCAM-1 mRNA expression was determined by RT-PCR and protein expression was detected by immunohistochemistry and/or western blotting. The data show that ICAM-1/VCAM-1 mRNA/protein expression was significantly enhanced in atherosclerotic aortas compared with normal diet group. Ox-LDL-induced ICAM-1/VCAM-1 mRNA/protein expression in U937 cells. Importantly, TSG significantly inhibited ICAM-1/VCAM-1 expression in atherosclerotic aortas in a dose-dependent manner. TSG-pretreatment also inhibited ICAM-1/VCAM-1 expression in Ox-LDL-induced U937 cells. Therefore, we concluded that TSG suppressed the expression of adhesion (ICAM-1/VCAM-1) molecules both in vivo (in aortic wall of dietary atherosclerotic rats) and in vitro (U937 cells).

Most cardiac diseases are associated with fibrosis. Calcineurin (CaN) is regulated by Ca(2+)/calmodulin (CaM). The CaN-NFAT (nuclear factor of activated T cell) pathway is involved in the process of cardiac diseases, such as cardiac hypertrophy, but its effect on myocardial fibrosis remains unclear. The present study investigates whether the CaN-NFAT pathway is involved in cardiac fibroblast (CF) proliferation induced by electrical field stimulation (EFS), which recently became a popular treatment for heart failure and cardiac tissue engineering. CF proliferation was evaluated by a cell survival assay (MTT) and cell counts. Myocardial fibrosis was assessed by collagen I and collagen III protein expression. Green fluorescent protein (GFP)-tagged NFAT was used to detect NFAT nuclear translocation. CF proliferation, myocardial fibrosis, CaN activity, and NFAT nuclear translocation were enhanced by EFS. More importantly, these effects were abolished by CaN inhibitors, dominant negative CaN (DN-CaN), and CaN gene silenced with siRNA. Furthermore, buffering intracellular Ca(2+) with BAPTA-AM and blocking Ca(2+) influx with nifedipine suppressed EFS-induced increase in intracellular Ca(2+) and CF proliferation. These results suggested that the CaN-NFAT pathway mediates CF proliferation, and that the CaN-NFAT pathway might be a possible therapeutic target for EFS-induced myocardial fibrosis and cardiac tissue engineering.

OBJECTIVE To develop a novel Escherichia coli cell surface display system by using C-terminally truncated NCgl1221 as the anchoring protein, which greatly enriched or optimized the bacterial displayed systems. METHODS We amplified the sequence of C-terminally truncated NCgl1221 and beta-amylase, and constructed the fusion expression vector. Then we transformed the recombinant plasmids PET-NA and PET-28a into Rosetta (DE3) pLysS. The fusion protein expression was induced by IPTG and identified by SDS-PAGE and Western blot analysis. The IPTG induced strains were immunostained and investigated by fluorescence microscope and flow cytometry to detect the displayed beta-amylase. Finally, we analyzed the activity of beta-amylase and starch hydrolization in order to determine whether the displayed beta-amylase has the activity or not. RESULTS The fusion protein was successfully expressed in E. coli, and the active beta-amylase was displayed on the cell surface by fusing it to the C terminus of the anchor. The recombinant strain displaying beta-amylase can utilize soluble starch in the medium. CONCLUSION A novel E. coli surface display system by using C-terminally truncated NCgl1221 as the anchor motif was successfully developed. The active enzyme with a molecular size of 56 kDa was displayed on E. coli by this system, which provided the basis for the application of the system in whole-cell biocatalyst or biosorbent.

Summary Cardiac fibroblasts (CF) have direct and potent effects on myocardial remodelling by proliferating, differentiating and secreting extracellular matrix proteins. Prolonged activation of CF leads to cardiac fibrosis and reduces myocardial contractile function. In previous studies we showed that 2,3,4′,5‐tetrahydroxystilbene‐2‐O‐β‐d‐glucoside (TSG) exerts cardiac protection, but the mechanism involved remains unclear. The aim of the present study was to evaluate the effects of TSG on angiotensin (Ang) II‐induced CF proliferation and to explore the underlying intracellular mechanisms. Angiotensin II (100 nmol/L)‐induced proliferation of rat neonatal fibroblasts was significantly inhibited by TSG (3–100 μmol/L), as evidenced by investigations of cell numbers and 5‐bromodeoxyuridine (BrdU) incorporation. In addition, 30 μmol/L TSG suppressed AngII‐induced expression of nuclear antigen, matrix metalloproteinase (MMP)‐2 and MMP‐9. Moreover, TSG attenuated AngII‐induced activation of mitogen‐activated protein kinase kinase (MEK) and extracellular signal‐regulated kinase (ERK) 1/2. Angiotensin II (100 nmol/L)‐induced generation of reactive oxygen species (ROS) was reduced by 30 μmol/L TSG, as was H2O2‐induced activation of ERK1/2. However, the MEK inhibitor 50 μmol/L PD98059 did not reduce ROS generation, although it did inhibit cell proliferation. There was a significant correlation between the inhibition of ERK1/2 activation and suppression of cell proliferation by TSG. However, there were no additive effects on either the inhibition of ERK1/2 or the suppression of cell proliferation following treatment of cells with both PD98059 and TSG. In conclusion, the results of the present study suggest that TSG inhibits ERK1/2 activation, likely via buffering of ROS, and consequently suppresses cell proliferation.

SUMMARY In coronary artery disease, the typical atheromatous plaque consists of a lipid core containing various inflammatory cells and a fibrous cap composed mostly of extracellular matrix. Both matrix metalloproteinases (MMPs) and inflammation are involved in the initiation of atherosclerotic plaques and plaque instability. 2,3,4¢,5‐Tetrahydroxystilbene‐2‐O‐b‐d‐glucoside (TSG) reduces the blood lipid content and prevents the atherosclerotic process, but the mechanism of action of TSG is unclear. The purpose of the present study was to test whether TSG can suppress MMP activation and inflammation in atherosclerotic rats. Sixty male Sprague‐Dawley rats were randomly divided into six groups. Atherosclerosis was induced by feeding rats a hyperlipidaemic diet; TSG (120, 60 or 30 mg/kg per day) was administered by oral gavage. After 12 weeks of treatment, rats were killed (ethyl carbamate 1200 mg/kg) and serum lipids, C‐reactive protein (CRP), interleukin (IL)‐6 and tumour necrosis factor (TNF)‐a were measured. Haematoxylin–eosin (H&E) staining was used to examine histopathological changes in the aorta. The mRNA and protein expression of MMPs were assayed by reverse transcription–polymerase chain reaction, immunohistochemistry and western blotting. Simvastatin (2 mg/kg per day) was administered as a positive control, whereas the vehicle (0.9% NaCl) group served as the untreated control. In the present study, TSG significantly and dose‐dependently attenuated the hyperlipidaemic diet‐induced alterations in serum lipid profile and increases in CRP, IL‐6 and TNF‐a levels. In addition, TSG normalized the structure of the aortic wall and suppressed the expression of MMP‐2 and MMP‐9 at both the mRNA and protein level in the rat aortic wall. In summary, TSG suppresses the expression of MMP‐2 and MMP‐9 and inhibits inflammation in the diet‐induced atherosclerotic rats.