9,9'-O-Isopropyllidene-isolariciresinol

Imaging of hypoxia-inducible factor 1alpha and septin 9 interaction by bimolecular fluorescence complementation in live cancer cells.[Pubmed:28380438]

Oncotarget. 2017 May 9;8(19):31830-31841.

Hypoxia-inducible factor 1 (HIF-1) is a major mediator of the hypoxic response involved in tumor progression. We had earlier described the interaction between septin 9 isoform 1 (SEPT9_i1) protein and the oxygen-regulated subunit, HIF-1alpha. SEPT9_i1 is a member of the conserved family of GTP-binding cytoskeleton septins. SEPT9_i1 stabilizes HIF-1alpha and facilitates its cytoplasmic-nuclear translocation. We utilized split yellow fluorescent protein (YFP) bimolecular fluorescence complementation (BiFC) methodology to monitor the interaction between HIF-1alpha and SEPT9_i1 in live cells. N-terminal (YN) and C-terminal (YC) split YFP chimeras with HIF-1alpha and SEPT9_i1 on both their amino and carboxyl termini were generated. HIF-1alpha and SEPT9_i1 chimeras were expressed in cancer cells and screened for functional complementation. SEPT9_i1-YN and YC-HIF-1alpha formed a long-lived highly stable complex upon interaction. The BiFC signal was increased in the presence of hypoxia-mimicking agents. In contrast, YC-DeltaHLH-HIF-1alpha chimera, which lacked the helix-loop-helix domain that is essential for the interaction with SEPT9_i1 as well as the expression of SEPT9_i1 252-379 amino acids fragment required for the interaction with HIF-1alpha, significantly reduced the BiFC signal. The signal was also reduced when cells were treated with 17-N-allylamino-17-demethoxygeldanamycin, an HSP90 inhibitor that inhibits HIF-1alpha. It was increased with fourchlorfenuron, a small molecule that increases the interaction between HIF-1alpha and SEPT9_i1. These results reconfirmed the interaction between HIF-1alpha and SEPT9_i1 that was imaged in live cells. This BiFC system represents a novel approach for studying the real-time interaction between these two proteins and will allow high-throughput drug screening to identity compounds that disrupt this interaction.

Growth performance of immunologically castrated pigs slaughtered at 5, 7, or 9 weeks after the second Improvest dose and fed diets containing corn dried distillers grains with solubles.[Pubmed:28380585]

J Anim Sci. 2017 Feb;95(2):806-819.

Growth performance of immunologically castrated (IC) pigs (863 total) was determined at increasing time intervals between the second Improvest (gonadotropin releasing factor analog-diphtheria toxoid conjugate; Zoetis Inc., Florham Park, NJ) dose and slaughter (TD) and with 4 different dried distillers grains with solubles (DDGS) feeding strategies (FS) in a 4 x 3 factorial arrangement of treatments. The feeding period was divided into 4 separate diet phases. Dietary treatments included 1) corn-soybean meal control diets (PCon), 2) a gradual decrease of dietary DDGS inclusion rate from 40%, 30%, 20%, and 10% in phases 1 to 4 (GD), respectively, 3) feeding 40% DDGS diets in phases 1 to 3 and removal of DDGS from the phase 4 diet (WD), and 4) feeding 40% DDGS diets in all 4 phases (NCon). Pigs received the second Improvest dose at 9 (TD9), 7 (TD7), or 5 (TD5) wk before slaughter. In each group, all pigs were slaughtered on the same day. There were no 3-way interactions among FS, TD, and week of feeding period for any measure of growth performance. Pigs fed PCon and WD had greater ( < 0.05) overall ADFI than pigs fed NCon, especially when slaughtered 9 wk after the second Improvest dose (2.45 and 2.44 vs. 2.31 +/- 0.08 kg/d, respectively). This response was partly due to withdrawing DDGS from the diet at 19 wk of age (WD), which led to a tendency ( < 0.10) for increased ADFI from the wk 19 to 21 interval to the wk 21 to 24 interval (3.26 vs. 3.51 +/- 0.09 kg/d, respectively). During the same time period, ADFI was unchanged ( > 0.05) in pigs fed PCon, GD, and NCon. Overall G:F was improved ( < 0.05) in TD5 pigs compared with TD9 pigs and tended ( < 0.10) to be improved compared with TD7 pigs. Final BW was similar among pigs fed GD, WD, and PCon (123.1, 122.3, and 125.3 kg, respectively), but pigs fed PCon and GD had greater ( < 0.05) BW than pigs fed NCon (120.0 kg). Throughout the growing-finishing period, BW was similar among TD treatments. The GD FS was more effective than the WD FS in maintaining overall G:F (0.424 and 0.414 +/- 0.005, respectively) and ADG (0.94 and 0.93 +/- 0.03 kg/d, respectively), which were similar ( > 0.05) to those of pigs fed PCon (0.427 +/- 0.005 and 0.96 +/- 0.03 kg/d, respectively). Growth performance of pigs fed GD more closely reflected that of pigs fed PCon than that of pigs fed WD. Delaying the second dose of Improvest from 9 to 5 wk before slaughter resulted in improved growth performance.

Caspase 12 degrades IkappaBalpha protein and enhances MMP-9 expression in human nasopharyngeal carcinoma cell invasion.[Pubmed:28380444]

Oncotarget. 2017 May 16;8(20):33515-33526.

Caspase-12 (Casp12), an inflammatory caspase, functions as a dominant-negative regulator of inflammatory responses and is associated with the signaling of apoptosis. However, the physiological function of Casp12 presented in cancer cells is still unclear. This study demonstrated that overexpression of Casp12 mediated IkappaBalpha degradation and significantly increased NF-kappaB activity. Exposure of human nasopharyngeal carcinoma (NPC) cells to phorbol-12-myristate-13-acetate (PMA) increased the levels of Casp12 and MMP-9 resulting in NPC cell invasion. Target suppression of Casp12 by small interfering RNA (siRNA) or an inhibitor of Casp12 markedly decreased the level of PMA-induced MMP-9 protein and cell invasion. Moreover, suppression of Casp12 significantly inhibited the basal activity of NF-kappaB and decreased the PMA-induced NF-kappaB reporter activity. The effect of Casp12 on NF-kappaB activation was indicated via the post-translational degradation of IkappaB. This study revealed that a critical role of Casp12 on the activation of NF-kappaB via IkappaBalpha degradation which provides a link between inflammatory and aggressive invasion in NPC cells.