MicroRNA 144 (miR-144), a small non-coding RNA, is frequently dysregulated in human several tumour progression, but its role and the underlying mechanisms in hepatocellular carcinoma (HCC) is poorly investigated. In the present study, the expression of miR-144 was firstly analysed in datasets derived from GSE21362 and TCGA, and then detected in HCC tissues and cell lines by quantitative RT-PCR (qRT-PCR) analysis. MiR-144 was shown to be significantly down-regulated in HCC tissues and cell lines. Subsequently, overexpression of miR-144 was transfected into HCC cell lines so as to investigate its biological function, including MTT, colony formation, and transwell assays. Gain of function assay revealed miR-144 remarkably inhibited cell proliferation, migration and invasion. In addition, bioinformatical analysis and luciferase reporter assay identified ZFX as a novel target of miR-144 in HCC cells, as confirmed by qRT-PCR and Western blot. Furthermore, ZFX was found to be significantly up-regulated using Oncomine database analysis. Loss of function assay further indicated knockdown of ZFX had similar effects of miR-144-mediated HCC cell proliferation and invasion. Therefore, miR-144 has been demonstrated to act as a tumour suppressor in HCC cell growth and motility by directly targeting ZFX, which implicates its potential applications in the development of HCC treatment.

Oxidative stress–induced vascular endothelial cell (VEC) injury is a major mechanism in the initiation and development of atherosclerosis. Lunasin, a soybean‐derived 43‐aa peptide, has been previously shown to possess potent antioxidant and anti‐inflammatory activities other than its established anticancer activities. This study investigated the effects of lunasin on protecting VECs from oxidative damage and inhibiting atherosclerotic plaque progression in apolipoprotein E–deficient (ApoE−/−) mice and explored its underlying mechanism. Biochemical and histologic analyses were performed by using EA.hy926 human VECs and a high‐fat diet (HFD) ApoE−/− mouse atherosclerosis model. Our data indicated that lunasin attenuated H2O2‐induced, mitochondria‐dependent endothelial apoptosis via down‐regulating Bax and up‐regulating Bcl‐2, inhibiting the mitochondrial depolarization, and reducing the release of cytochrome c, as well as decreasing the activation of casρase‐9 and caspase‐3 in vitro and in vivo. Mechanic studies showed that lunasin significantly up‐regulated heme oxygenase‐1 via the PI3K/Akt/nuclear factor erythroid 2–related factor 2/antioxidant response element pathway, and reduced H2O2‐induced ROS production in VECs, thereby attenuating oxidant‐induced endothelial injury and inhibiting atherosclerotic plaque progression in ApoE−/− mice. In conclusion, our in vitro and in vivo data suggest that lunasin protects VECs from oxidative damage by enhancing heme oxygenase‐1 expression via activation of the PI3K/Akt/ nuclear factor erythroid 2–related factor 2/antioxidant response element pathway and inhibiting mitochondria‐dependent apoptosis, thereby effectively attenuating atherosclerosis in HFD‐fed ApoE−/− mice. Lunasin may act as a potential therapeutic agent for the prevention and treatment of atherosclerosis.—Gu, L., Ye, P., Li, H., Wang, Y., Xu, Y., Tian, Q., Lei, G., Zhao, C., Gao, Z., Zhao, W., Tan, S. Lunasin attenuates oxidant‐induced endothelial injury and inhibits atherosclerotic plaque progression in ApoE−/− mice by up‐regulating heme oxygenase‐1 via PI3K/Akt/ Nrf2/ARE pathway. FASEB J. 33, 4836–4850 (2019). http://www.fasebj.org

Background Hepatocellular carcinoma (HCC) occurs frequently in China, with high morbidity and mortality. Cell division cycle 20 homolog ( CDC20 ) is reportedly related to many cancers. In this study, we discuss a potential link of CDC20 expression to HCC patients’ prognoses. Material/Methods Quantitative real-time polymerase chain reaction (qRT-PCR) was performed to assess CDC20 expression in HCC and the paired noncancerous tissues. Chi-square analysis was used to assess potential association of CDC20 expression with clinicopathologic profiles among HCC patients. The overall survival for HCC patients with different CDC20 expressions was assessed using the Kaplan-Meier method. To evaluate the prognostic value for HCC patients, Cox regression analyses were performed. Results The expression of CDC20 was elevated among HCC specimens compared with adjacent noncancerous ones ( P <0.05). The expression of CDC20 was significantly related to differentiation ( P <0.001), tumor node metastasis stage ( P <0.001), and lymphatic metastasis ( P <0.001). Moreover, HCC patients with high CDC20 expression had dismal overall survival rates compared with low CDC20 expression ( P <0.05). CDC20 alone could forecast HCC prognoses according to multivariable Cox regression analysis (hazard ratio=2.354, 95% confidence interval=1.177–4.709, P =0.016). Conclusions Overexpressed CDC20 may act as a reliable biomarker for dismal prognoses among HCC patients.

BackgroundIt has been long-known that endometrium-secreted cytokines play a critical role during embryo implantation. However, whether cytokines secreted from the embryo are relevant to the process of embryo implantation remains unclear. MethodsThe concentration of cytokines in embryo culture medium was tested using a newly developed, highsensitivity single-cell proteomic platform and evaluated in comparison to embryo quality and clinical outcome. The effect of TNF-α on embryo and endometrium Ishikawa cells was investigated using immuno uorescence staining, CCK-8 assay, TUNEL staining, and RT-qPCR. ResultsOf the 10 cytokines measured, only TNF-α concentration was signi cantly higher in the group with embryo implantation failure. Immuno uorescence staining showed that the expression of TNF-α was unevenly distributed in blastocysts, and the expression level was signi cantly correlated with the blastocyst inner cell mass (ICM) quality score. Gene pro ling showed that addition of TNF-α led to increased expression of tumor necrosis factor receptor 1 (TNFR1) and apoptosis-related genes and that this could be inhibited by the TNF-α receptor inhibitor entanecept (ETA). In addition, an increased expression of water and ion channels, including AQP3, CFTR, ENaCA and CRISP2 was also observed which could also be inhibited by ETA. ConclusionsOur results show that higher embryo-secreted TNF-α levels are associated with implantation failure through activation of TNF-α receptor, and TNF-α may be an independent predictor for pre-transfer assessment of the embryo development potential in IVF patients.

Fusion expression provides an effective means for the biosynthesis of longer peptides in Escherichia coli. However, the commonly used fusion tags are primarily suitable for laboratory scale applications due to the high cost of commercial affinity resins. Herein, a novel approach exploiting hirudin as a multipurpose fusion tag in combination with tobacco etch virus (TEV) protease cleavage has been developed for the efficient and cost-effective production of a 43-amino acid model peptide lunasin in E. coli at preparative scale. A fusion gene which allows for lunasin to be N-terminally fused to the C-terminus of hirudin through a flexible linker comprising a TEV protease cleavage site was designed and cloned in a secretion vector pTASH. By cultivation in a 7-L bioreactor, the fusion protein was excreted into the culture medium at a high yield of ~380 mg/L, which was conveniently recovered and purified by inexpensive HP20 hydrophobic chromatography at a recovery rate of ~80%. After polishing and cleavage with TEV protease, the finally purified lunasin was obtained with ≥95% purity and yield of ~86 mg/L culture medium. Conclusively, this hirudin tagging strategy is powerful in the production of lunasin and could be applicable for the production of other peptides at preparative scale.