Identification of glutamyl-prolyl-tRNA synthetase as a new therapeutic target in hepatocellular carcinoma via a novel bioinformatic approach.

Background: Hepatocellular carcinoma (HCC) has a high incidence, and current treatments are ineffective. We aimed to explore potential diagnostic and prognostic biomarkers for HCC by conducting bioinformatics analysis on genomic and proteomic data. Methods: Genome and proteome data were downloaded from The Cancer Genome Atlas (TCGA) and ProteomeXchange databases, respectively. Differentially expressed genes was determined using limma package. Functional enrichment analysis was conducted by Database for Annotation, Visualization, and Integrated Discovery (DAVID). Protein-protein analysis was established by STRING dataset. Using Cytoscope for network visualization and CytoHubba for hub gene identification. The gene mRNA and protein levels were validated using GEPIA and HPA, as well as RT-qPCR and Western blot. Results: A total of 127 up-regulated and 80 down-regulated common DEGPs were identified between the genomic and proteomic data, Mining 10 key genes/proteins(ACLY, ACACB, EPRS, CAD, HSPA4, ACACA, MTHFD1, DMGDH, ALDH2, and GLDC) through protein interaction networks. in addition, Glutamyl-prolyl-tRNA synthetase (EPRS) was highlighted as an HCC biomarker that is negatively correlated with survival. Differential EPRS expression analysis in HCC and paracancerous tissues showed that EPRS expression was elevated in HCC. RT-qPCR and Western blot analysis results showed that EPRS expression was upregulated in HCC cells. Conclusions: Our results suggest that EPRS is a potential therapeutic target for inhibiting HCC tumorigenesis and progression.

CircKIF4A combines EIF4A3 to stabilize SDC1 expression to activate c-src/FAK and promotes TNBC progression.

Triple-negative breast cancer (TNBC) is recognized for its poor prognosis and limited options for treatment. Circular RNA KIF4A (circKIF4A) was documented to be abnormally overexpressed in TNBC and was correlated with a poor survival rate. The objective of this study is to further examine the functional role of circKIF4A and its underlying mechanism. CircKIF4A was significantly upregulated in TNBC and the knockdown of circKIF4A suppressed TNBC cell proliferation, migration, and invasion. CircKIF4A was directly bound to EIF4A3, which interacted with SDC1. Knockdown of circKIF4A reduced interaction between EIF4A3 and SDC1 as well as SDC1 mRNA stability. SDC1 activated the c-src/FAK signaling pathways and finally promoted TNBC progression. circKIF4A induced TNBC progress in the in vivo mouse model via SDC1. CircKIF4A interacts with EIF4A3 to stabilize SDC1 mRNA, which activates the c-src/FAK signaling pathways and promotes TNBC progression. This may provide a potential therapy for TNBC treatment.

Azoreductase and Target Simultaneously Activated Fluorescent Monitoring for Cytochrome c Release under Hypoxia.

Hypoxia-induced cell apoptosis is closely related to degenerative diseases, autoimmune disorders, and tumor disease. In the process of apoptosis, the release of cytochrome c (Cyt c) is deemed to be a critical factor of the intrinsic pathway. Strategies for tracking Cyt c release in living cells based on the subcellular localization have been proposed recently. However, they are inherently lack of specificity for distinguishing the release of Cyt c in apoptotic process induced by hypoxia from other stimulus. In this paper, an azoreductase and target simultaneously activated fluorescent aptameric nanosensor integrating gold nanoparticles (AuNPs) and Cyt c-targeted aptamer-consisted double-stranded DNA hybridization complex (DSDHC) was proposed. It is worth noting that the employment of azobenzene moiety labeled on the DSDHC first ensured the aptameric nanosensor could be conjugated to the surface of AuNPs and then specifically reduced by hypoxia-related azoreductase. Upon Cyt c released from mitochondrion under hypoxia, the competitive displacement of Cyt c subsequently activated the fluorescence of the aptameric nanosensor and the fluorescence enhancement depended principally on the content of Cyt c release. Inspired by this, a new strategy for quantitative analysis and in situ imaging of Cyt c under hypoxic condition was proposed. The high spatial resolution monitoring of the dynamics of Cyt c release under hypoxia will offer a potentially rich opportunity to understand the apoptotic mechanism under hypoxic conditions, thus further facilitating risk assessment and risk reduction for hypoxic environments.

[Effect of cordyceps sinensis extract on Klotho expression and apoptosis in renal tubular epithelial cells induced by angiotensin II].

OBJECTIVE: To investigate the effect of cordyceps sinensis (CS) extract and losartan (Los) on the expression of Klotho (Kl), P53, P21, and apoptosis in renal tubular epithelial cell NRK-52E induced by angiotensin II (Ang II), and to elucidate its therapeutical mechanism in Ang II induced renal tubular epithelial cell apoptosis. METHODS: NRK-52E cells were incubated with CS with or without Ang II for 24 hours. Experimental groups were divided according to the increasing concentrations of CS:0 (serving as controls), 5, 10, 20, 40, and 80 mg/L. The optimal concentration of CS was selected and cells were divided into 5 groups: controls, Ang II (1*10(-8) mol/L), Ang II (1*10(-8) mol/L)+CS (40 mg/L), Ang II (1*10(-8) mol/L)+Los (1*10(-5) mol/L), and Ang II (1*10(-8) mol/L)+CS (40 mg/L)+Los (1*10(-5) mol/L). After 24 hours, cell proliferation was evaluated by MTT assay. The mRNA and protein expression of Kl, P53 and P21 were measured by RT-PCR. Activity of caspase-3 was evaluated by caspase-3 activity assay Kit. Cell apoptosis was determined by Annexin V-FITC/PI double staining and flow cytometry. RESULTS: Certain concentrations of CS promoted the proliferation of NRK-52E cells and increased cells proliferation inhibited by Ang II (P<0.01 or P<0.05 ). Ang II significantly down-regulated the mRNA and protein expression of Kl, and up-regulated the levels of P53 and P21. Caspase-3 activity and apoptotic rates were decreased, too (all P values<0.01). CS or/and Los significantly increased the expression of Kl mRNA and protein down-regulated by Ang II, decreased P53 mRNA and protein expression, P21 mRNA and protein expression,and inhibited caspase-3 activity and apoptotic rates(all P values<0.05). No cooperative effects were observed in the two drugs (P>0.05). CONCLUSION: CS can increase the expression of Kl down-regulated by Ang II, decrease P53 and P21 expression and caspase-3 activity, and reduce Ang II induced NRK-52E cell apoptosis, which may be part of its mechanism of the protective effects on hypertensive renal damage.