Cucurbitacin induces autophagy through mitochondrial ROS production which counteracts to limit caspase-dependent apoptosis

Targeted disruption of STAT3 function has proven to be a useful cancer therapeutic approach by inducing apoptotic cell death. Cucurbitacin is currently under development as a small molecule of STAT3 inhibitor to trigger cell death in many cancers. Here, we systematically studied the molecular mechanisms underlying cucurbitacin-induced cell death, in particular the involvement of autophagy. Treatment with cucurbitacin resulted in non-apoptotic cell death in a caspase-independent manner. Notably, cucurbitacin enhanced excessive conversion of lipidated LC3 (LC3-II) and accumulation of autophagosomes in many cell types. Such autophagy and cell death induced by cucurbitacin were independent of its ability to inhibit STAT3 function, but mainly mediated by enhanced production of mitochondrial-derived reactive oxygen species (ROS), and subsequently activation of extracellular signal-regulated kinase (ERK) and c-jun NH2-terminal kinase (JNK). Interestingly, both the autophagy inhibitor wortmannin and knockdown of Atg5 or Beclin 1 failed to rescue the cells from cucurbitacin-induced cell death, as suppression of autophagy induced the mode of cell death to shift from autophagic cell death to caspase-dependent apoptosis. Thus the present study provides new insights into the molecular mechanisms underlying cucurbitacin-mediated cell death and supports cucurbitacin as a potential anti-cancer drug through modulating the balance between autophagic and apoptotic modes of cell death.     

抗速灭威单链抗体基因在巴斯德毕赤酵母中的表达及性质研究

选用巴斯德毕赤酵母(Pichia pastoris)系统表达特异性抗速灭威单链抗体(scFv)基因,以为速灭威特异性抗体的大量制备奠定基础。设计引物扩增阳性克隆scFv基因,亚克隆至表达载体pPICZαC,获得重组酵母表达质粒pPICZαC-scFv,线性化pPICZαC-scFv并高效电转P.pastoris(X-33),对转化子进行抗性梯度筛选得到一株高效表达的X-33-Pp-SMW-12-6菌株。对获得的菌株先后进行表达条件的优化、优化条件下的诱导表达及单链抗体性质研究。结果表明:P.pastoris-scFv的质量接近其亲本E.coli-scFv的质量,X-33-Pp-SMW12-6在优化条件下的表达产量达28mg/L,比未优化前的产量提高了约8mg/L,经纯化后抗体纯度可达85%以上。因此,利用P.pastoris表达系统制备抗速灭威单链抗体比细菌表达系统更有效、更经济。     

Molecular cloning, distribution and ontogenetic expression of the oligopeptide transporter PepT1 mRNA in Tibetan suckling piglets

The gene encoding the oligopeptide transporter PepT1 (HGMW-approved gene symbol SLC15A1) from Tibetan porcine intestine was cloned. The open reading frame of this cDNA encodes 708 deduced amino acid residues that show high sequence similarity with its ovine and bovine counterparts. The putative protein has 12 putative transmembrane domains, including many structural features that are highly conserved among the vertebrate orthologs. PepT1 mRNA expression can be detected in duodenum, jejunum and ileum from Tibetan pigs at 28 days by RT-PCR. Real-time PCR analysis indicated that the jejunum had the highest expression of PepT1 when compared with the duodenum and ileum. PepT1 mRNA expression in the duodenum and proximal jejunum increases continuously from day 1 to day 14: expression was highest at day14 (P < 0.01) and then decreased gradually from day 21 to day 35. Our findings show that PepT1 mRNA expression in the distal jejunum increased gradually with age in suckling Tibetan piglet, and this may have important implications for amino acid and protein nutrition in young animals.     

Androgen receptor coregulator ARA267-α interacts with death receptor-6 revealed by the yeast two-hybrid

ARA267-α is a newly identified androgen receptor coactivator. In order to further elucidate its precise role in cells, using the ARA267-α fragment containing four PHD and one SET conserved domains as bait we revealed an ARA267-α-PHD-SET-interacting protein, death receptor-6 (DR6), in the yeast two-hybrid screening. DR6 is the member of TNF receptor family and has a death domain in its intracellular cytoplasmic portion (DR6cp) to mediate the cell apoptosis. The interaction between ARA267-α-PHD-SET and DR6cp was confirmed in vitro and in vivo. Our finding implied that androgen signaling pathway might cross talk with apoptosis signaling pathway through the interaction between ARA267-α and DR6.     

Proteomic Analysis of Shoot Tips from Two Alfalfa Cultivars with Different Florescence

Plant Molecular Biology Reporter - Flowering is an indispensable biological process for the complete life cycle of angiosperms, crucial to the regeneration of plants and the continuation of...     

Effects of L-serine supplementation on the daily rhythms of growth hormone and corticosterone concentrations in mice

The impact of L-serine on the daily rhythms of growth hormone (GH) and corticosterone remains unknown. We explored whether the daily rhythms of these hormones were affected by L-serine supplementation as well as the supplementation time. The results showed that plasma GH concentration at Zeitgeber time (ZT)4 and 8 were significantly increased by L-serine supplementation at ZT22, while the diurnal rhythms peaks of plasma corticosterone at ZT12 were suppressed by L-serine supplementation at ZT10. After the supplementation was stopped, the effects of L-serine on the diurnal rhythms of plasma GH and corticosterone lasted for 2 days then they were fading on day 4. L-serine concentrations in plasma and hypothalamus after supplementation at ZT22 was lower than those after supplementation at ZT10. In conclusion, L-serine modulates the daily rhythms of GH and corticosterone depending on its supplementation time. The modulation effect might be association with the daily rhythms of L-serine metabolism.