ZNF146 regulates cell cycle progression via TFDP1 and DEPDC1B in ovarian cancer cells.

In brief: Aberration in cell cycle progression is one of the essential mechanisms underlying tumorigenesis, making regulators of cell cycle reasonable anti-cancer therapeutic targets. Here, we dissected the regulatory mechanism involving the novel axis ZNF146/TFDP1/DEPDC1B in the cell cycle in ovarian cancer. Abstract: Ovarian cancer (OC) is the third most common kind of gynecological tumor, in addition to being the most lethal. Transcription factor Dp-1 (TFDP1) functions as a binding partner for E2F transcription factors, and its target genes include those involved in DNA synthesis, cell cycle, and apoptosis. However, the regulatory role of TFDP1 in OC remains incompletely understood. This study aimed to investigate the role and mechanism of TFDP1 in OC. TFDP1 was highly expressed in the ovarian epithelial tissues of OC patients, and the expression of TFDP1 in OC cells was higher than that in normal ovarian epithelial cells. Silencing of TFDP1 inhibited the biological activity of OC cells and hindered cell cycle entry. Zinc finger protein 146 (ZNF146) knockdown induced cell cycle arrest at the G0/G1 phase and tumor growth by blocking TFDP1 transcription, which was overturned by ectopic expression of TFDP1. TFDP1 stimulated DEP domain-containing protein 1B (DEPDC1B) expression through transcriptional activation. DEPDC1B increased the proportion of OC cells in the G2/M phase and potentiated tumor malignant progression in nude mice inhibited by sh-ZNF146. Taken together, these findings demonstrate that ZNF146 participates in TFDP1/DEPDC1B activation and plays a vital role in the cell cycle in OC.

Molecular Surgery at Microporous MOF for Mesopore Generation and Renovation.

Hierarchically porous MOFs (HP-MOFs) present advantageous synergism of micro- and mesopore but challenging in synthetic control at molecular scale. Herein, we present the first example of reversible and controllable mesopore generation and renovation in a microporous MOF of HKUST-1 via synthetic manipulation at molecular scale. An ammonia-gas etching strategy is proposed to create mesopores in carboxylate-based microporous MOFs and thus produce HP-MOFs. Gas-phase etching ensures uniform mesopore formation inside the MOF crystals via plane-oriented cutting the carboxylate-metal bonds off without affecting the crystal size and morphology. The mesopore size is controlled by the etching temperature, while the mesopore volume could be tuned by adjusting etchant pressure. The generated mesopores could be renovated using MOF precursors solutions so that to achieve controllable mesopore generation/closure, and encapsulation of the adsorbed molecules. This work demonstrates a powerful protocol for precisely tailoring and tuning the properties of MOF materials at molecular scale.

Engineered antibody CH2 domains binding to nucleolin: Isolation, characterization and improvement of aggregation.

Smaller recombinant antibody fragments are now emerging as alternatives of conventional antibodies. Especially, immunoglobulin (Ig) constant CH2 domain and engineered CH2 with improved stability are promising as scaffolds for selection of specific binders to various antigens. We constructed a yeast display library based on an engineered human IgG1 CH2 scaffold with diversified loop regions. A group of CH2 binders were isolated from this yeast display library by panning against nucleolin, which is a tumor-associated antigen involved in cell proliferation, tumor cell growth and angiogenesis. Out of 20 mutants, we selected 3 clones exhibiting relatively high affinities to nucleolin on yeasts. However, recombinant CH2 mutants aggregated when they were expressed. To find the mechanism of the aggregation, we employed computational prediction approaches through structural homology models of CH2 binders. The analysis of potential aggregation prone regions (APRs) and solvent accessible surface areas (ASAs) indicated two hydrophobic residues, Val264 and Leu309, in the ß-sheet, in which replacement of both charged residues led to significant decrease of the protein aggregation. The newly identified CH2 binders could be improved to use as candidate therapeutics or research reagents in the future.

Multivariate prognostic study on node-positive gastric cancer: is tumor size a prognostic indicator?

BACKGROUND/AIMS: We analyzed the clinicopathological factors of patients with node-positive gastric cancer, evaluated the prognostic factors associated with long-term survival and clarified the effect of tumor size on long-term survival. METHODOLOGY: The study included 591 patients who underwent curative resection for node-positive gastric cancer. Clinicopathological prognostic variables were evaluated as predictors of long-term survival by univariate and multivariate analyses. RESULTS: The 5-year survival rate was influenced by tumor size, tumor location, depth on invasion, level of lymph node metastasis, Borrmann classification, histological type, liver metastasis, peritoneal dissemination and disease stage. Of these, independent prognostic factors were depth on invasion and lymph node metastasis. Tumor size is an influence but not independent factor for the prediction of long-term survival in patients with node-positive gastric cancer. CONCLUSIONS: In patients with node-positive gastric cancer, two independent prognostic factors were depth on invasion and the status of lymph node metastasis.

Coordination-Assistant Chiral Agent Anchoring on Amphiphilic Graphitic Phase Carbon Nitride Membrane for Multiple Molecular Separation.

Membranes composed of two-dimensional (2D) materials suffer from low stability and structural swelling and are usually restricted to applications in aqueous systems. Among various 2D materials, graphitic phase carbon nitride (GCN, g-C3N4) has shown great application potential owing to its structural tunability. Herein, we develop a coordination-assisted strategy to regulate the GCN layer spacing and chemical environment via copper ion (Cu2+) coordination-assisted intercalation of enantiopure (1S,2S)-(-)-1,2-diphenyl-1,2-ethanediamine (DPE) between GCN nanosheets. The obtained GCN-Cu-DPE membrane is continuous and intact, free of cracks and pinholes, stable under acidic and alkaline conditions, and exhibits water permeability above 215 L m-2 h-1 bar-1 and a high rejection rate to dye molecules. The membrane is amphiphilicity and thus allows both polar solvent (water) and nonpolar solvent (hexane) to freely pass through. Remarkably, the permeation rate is proportional to the viscosity of the solvent. Benefiting from the chiral space between nanosheets, the GCN-Cu-DPE membrane shows selective permeation of aspartic acid racemate in aqueous systems and limonene racemate in the organic phase. Our work demonstrates a general and promising strategy for chiral membrane fabrication toward high-value-added chiral separation, especially in the pharmaceutical industry.

Knockdown of TRIM44 inhibits the progression of ovarian cancer and is related to the FOXM1-EZH2 signaling pathway.

Background: Tripartite motif-containing protein 44 (TRIM44) was recently identified as a novel oncogene that is overexpressed in several types of human cancers. However, the biological functions of TRIM44 in epithelial ovarian cancer (EOC) remain unclear. Here, we aimed to investigate the role of TRIM44 in EOC and its clinical implications. Methods: TRIM44 was knocked down using shRNA transfection. In vitro proliferation, invasion, migration and apoptosis of ovarian cancer (OC) cells were detected by CCK8, colony formation assay, Transwell inserts and flow cytometry analysis. The growth ability of xenograft tumors was examined in vivo in a nude mouse metastatic tumor model. Finally, we performed gene chip analysis and ingenuity pathway analysis (IPA) to analyze the potential gene network. Results: High expression of TRIM44 was observed in EOC tissues. Knockdown of TRIM44 expression substantially suppressed the proliferation, migration, invasion and colony-forming ability of EOC cells in vitro and attenuated tumor growth in vivo. Mechanistic studies revealed that silencing TRIM44 dramatically downregulated the expression of FOXM1, EZH2, CCNE2, CCND3 and BIRC5 in EOC cells, at least in part through inactivation of the FOXM1-EZH2 signaling pathway. Conclusions: Collectively, these data suggest that downregulation of TRIM44 inhibits the progression of EOC through suppression of the FOXM1-EZH2 signaling pathway. These results provide novel insight into the role of TRIM44 in tumorigenesis and suggest that it could be a potential therapeutic target for ovarian carcinoma.

Efficient Photo-Thermo-Electric Conversion Using Polyoxovanadate in Ionic Liquid for Low-Grade Heat Utilization.

A large fraction of energy, including solar energy, is dissipated into ambient atmosphere as low-grade waste heat. Efficient utilization of such energy is critical to address the current energy crisis and global warming issue. Herein, the efficient near-IR (NIR)-photothermal, thermoelectric, and thus photo-thermo-electric conversion of polyoxovanadate compound {[Ni(1,10-phenanthroline)3 ][V14 O34 Cl]Cl, NiV14 } in ionic liquid was achieved. The solution displayed a NIR-photothermal efficiency of 16.04 and 23.43 % at 808 and 1064 nm, respectively. Taking advantage of the synergetic thermodiffusive and thermogalvanic effects of various ion species in NiV14 solution, an open circuit voltage of approximately 0.45 V was obtained at ΔT=70 K generated by physical heating or NIR irradiation, indicating a large Seebeck coefficient of 6.38 mV K-1 and an optimized thermal power at 1.2 W m-2 . The polyoxovanadate-ionic liquid system offers a new platform for efficiently utilizing not only low-grade thermal energy but also solar energy for electricity generation.

[Exploration on the increased sensitivity to docetaxel and reversing mechanism of drug resistance of antisense survivin RNA in gastric cancer cell line SGC7901 cells].

OBJECTIVE: To explore the effects of survivin antisense RNA on apoptosis and chemosensitivity to docetaxel in gastric cancer line SGC7901 cells, and its relation to mdr-1. METHODS: survivin antisense eukaryotic vector anti-pcDNA3-svv was transfected into SGC7901 cells by electorporation and positive clone was screened out. survivin protein and mdr-1 mRNA were determined by Western blot and RT-PCR. Apoptosis-inducing effect was examined by electron microscopy. Sensitivity to docetaxel was examined by MTT. Expression of mdr-1 and survivin mRNA were detected in the SGC7901 cells after drug-resisitance induction. RESULTS: The expression of survivin protein of SGC7901 cells after transfection reduced significantly than that of non-transfected cells. MDR indexes of transfection group and non-transfection group were 0.196 +/- 0.013 and 3.126 +/- 0.019, respectively. The IC50 of transfection group to docetaxel was (16.7 +/- 1.98) microg/L and non-transfection group was (55.7 +/- 1. 89) microg/L, with a statistically significant difference. Expression of survivin mRNA in drug-resistant cells decreased along with the decreasing of mdr-1. CONCLUSION: Antisense surivivin RNA can induce apoptosis in gastric cancer cells and increase sensitivity to docetaxel. The reversing mechanism of drug resistance is related with decreasing of mdr-1.

Triptolide inhibits viability and induces apoptosis in liver cancer cells through activation of the tumor suppressor gene p53.

The present study investigated the effect of triptolide on viability and apoptosis along with underlying mechanism in liver cancer cells. CCK-8 assay showed that triptolide treatment for 48 h significantly reduced the viability of HepG2 and QSG7701 cells at 50 µM concentration. Annexin V-FITC and propidium iodide staining showed that triptolide treatment of HepG2 cells at 50 µM concentrations induced apoptosis in 56.45% cells compared to only 2.36% cells in the control cultures. Western blot assay showed that treatment of HepG2 cells with 50 µM concentration of triptolide significantly induced phosphorylation of p53 in a 2 h-treatment. Phosphorylation of histone H2A.X indicator of DNA damage was induced by triptolide treatment after 12 h in HepG2 cells. The level of nuclear p53 in a 6 h-treatment with 0, 10, 20, 30, 40 and 50 µM concentration of triptolide was found to be 15.3, 19.6, 28.5, 43.7, 63.8 and 91.5%, respectively. Treatment of HepG2 cells with triptolide at 50 µM concentration caused a significant increase in the binding potential of p53 to DNA. Triptolide treatment of HepG2 cells caused a significant increase in the expression of p21, Bax and DR5 genes in HepG2 cells. It also increased the expression of miR-34b and miR-34c in HepG2 cells markedly. Treatment of HepG2 cells with p53 inhibitor, pifithrin-α prior to incubation with triptolide significantly prevented induction of cell apoptosis. Suppression of p53 expression by siRNA inhibited the expression of p53 as well as its target genes along with the prevention of apoptosis induction. In conclusion, triptolide inhibits viability and induces apoptosis in liver cancer cells through activation of the tumor suppressor gene p53. Thus, triptolide can be used for the treatment of liver cancer.

A potential prognostic long non-coding RNA signature to predict metastasis-free survival of breast cancer patients.

Long non-coding RNAs (lncRNAs) have been implicated in a variety of biological processes, and dysregulated lncRNAs have demonstrated potential roles as biomarkers and therapeutic targets for cancer prognosis and treatment. In this study, by repurposing microarray probes, we analyzed lncRNA expression profiles of 916 breast cancer patients from the Gene Expression Omnibus (GEO). Nine lncRNAs were identified to be significantly associated with metastasis-free survival (MFS) in the training dataset of 254 patients using the Cox proportional hazards regression model. These nine lncRNAs were then combined to form a single prognostic signature for predicting metastatic risk in breast cancer patients that was able to classify patients in the training dataset into high- and low-risk subgroups with significantly different MFSs (median 2.4 years versus 3.0 years, log-rank test p < 0.001). This nine-lncRNA signature was similarly effective for prognosis in a testing dataset and two independent datasets. Further analysis showed that the predictive ability of the signature was independent of clinical variables, including age, ER status, ESR1 status and ERBB2 status. Our results indicated that lncRNA signature could be a useful prognostic marker to predict metastatic risk in breast cancer patients and may improve upon our understanding of the molecular mechanisms underlying breast cancer metastasis.

[Survivin antisense RNA induces apoptosis and sensitizes ovarian cancer cell line SKOV3 to docetaxel].

BACKGROUND & OBJECTIVE: Survivin gene overexpresses in a variety of human tumors, and plays an important role in cell apoptosis and drug resistance of tumors. This study was designed to establish Survivin antisense RNA, and explore its effects on apoptosis of ovarian cancer cell line SKOV3 and sensitivity to docetaxel. METHODS: Survivin antisense eukaryotic expression vector anti-pcDNA3-svv was established, and transfected into SKOV3 cells by electroperforation. Positive clones (SKOV3-SVVanti) were screened out. Survivin mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR); Survivin protein was detected by Western blot. The effect of Survivin antisense RNA on apoptosis of SKOV3 cells was measured by flow cytometry and observed under electron microscope; its effect on sensitivity of SKOV3 cells to docetaxel was examined by MTT assay. RESULTS: The mRNA and protein levels of Survivin were obviously lower in SKOV3-SVVanti cells than in control cells. Terminal apoptosis changes were observed under electron microscope after transfection. The apoptosis rate was 19%. The 50% inhibitory concentration (IC50) of docetaxel was significantly lower for SKOV3-SVVanti cells than for control cells [(13.3+/-2.2) ng/ml vs. (53.2+/-2.4) ng/ml, P<0.05]. CONCLUSION: Surivivin antisense RNA can induce apoptosis of SKOV3 cells, and sensitize SKOV3 cells to docetaxel.