Sh-MARCH8 Inhibits Tumorigenesis via PI3K Pathway in Gastric Cancer

Background/Aims: To identify new treatment strategies for gastric cancer and to elucidate the mechanism underlying its pathophysiology, we transfected sh-MARCH8 into the human gastric cancer cell lines MKN-45 and AGS to investigate the roles of MARCH8 in gastric cancer. Methods: We used genetic engineering to construct the sh-MARCH8 interference plasmid and transfected it into gastric cancer cells. Colony formation assays and cell viability measurements were performed to detect the viability and proliferation of cancer cells. Wound healing assays were performed to estimate the migration and proliferation rates of the cells. Cell invasion assays were used to estimate the invasive abilities of the cells. Cell apoptosis analysis was performed by using flowing cytometry. Western blot analysis was performed to estimate the expression levels of proteins. Statistical analysis was performed using the SPSS 18.0 software. Student’s t-test was used to determine the significance of all pairwise comparisons of interest. Results: We observed that the transfection of sh-MARCH8 inhibited the survival and proliferation of MKN-45 and AGS cells. The migration and invasion of the MKN-45 and AGS cells were significantly decreased, and apoptosis was induced in comparison with the control cells. These results were further confirmed by data showing that sh-MARCH8 increased the BAX/BCL2 ratio in MKN-45 and AGS cells. We also observed that sh-MARCH8 inactivated the PI3K and ß-catenin stat3 signaling pathways by changing protein expression levels or the phosphorylation of related proteins. Conclusion: These data suggested that sh-March8 reduced viability and induced apoptosis of the MKN-45 and AGS cells through the PI3K and ß-catenin stat3 signaling pathways. Taken together, our data revealed that transfection of sh-MARCH8 into the MKN-45 and AGS gastric cancer cell lines inhibited their growth, and this approach may be useful as a novel strategy for gastric cancer therapy

HS-GC-IMS and PCA to Characterize the Volatile Flavor Compounds in Three Sweet Cherry Cultivars and Their Wines in China

The aim of this research was to characterize differences and sources of volatile flavor compounds by using headspace-gas chromatography-ion mobility spectrometry (HS-GC-IMS) and principal component analysis (PCA). Three sweet cherry fruits from different cultivars (cv. Tie, Van, and Lap) and their wines that were produced by the same yeast were detected. The results showed that 27 flavor compounds were identified in cherry fruits, including 10 alcohols, 7 esters, 7 aldehydes, 2 ketones, and 1 organic acid. Twenty-three flavor compounds were identified in cherry wines, including nine esters, eight alcohols, three aldehydes, two organic acids, and one ketone. In cherry fruits, aldehydes, several alcohols, and one ketone were the most prevalent in cv. Tie, and the majority of esters and alcohols in cv. Van. After fermentation, ethanol, butanol, butanal, ethyl propionate, propionaldehyde, 3-hydroxy-2-butanone, and acetic acid increased, whereas 1-hexanol, 3-methyl-3-buten-1-ol, 1-penten-3-ol, ethyl acetate, methyl acetate, (E)-2-hexenal and hexanal decreased. Few differences were detected in the type and content of volatile compounds in cherry wines from cv. Tieton (WT) and cv. Van (WV). Almost all aldehydes are derived from cherry fruits, which cannot be produced during wine-making, and other volatile compounds are almost all produced by saccharomyces cerevisiae. The volatile compounds of cherry wines were determined by row materials and fermentation cultures. Flavor fingerprints were established by HS-GC-IMS and PCA, which provided a theoretical foundation for the evaluation and improvement of flavor quality in cherry wine-making

Strengthening Gastric Cancer Therapy by Trastuzumab-Conjugated Nanoparticles with Simultaneous Encapsulation of Anti-MiR-21 and 5-Fluorouridine

Background/Aims: MicroRNA-21 is an oncogenic miR (oncomiR) frequently elevated in gastric cancer (GC). Overexpression of miR-21 decreases the sensitivity of GC cells to 5-fluorouridine (5-Fu) and trastuzumab, a humanized monoclonal antibody targeting human epidermal growth factor receptor 2 (HER2). Receptor-mediated endocytosis plays a crucial role in the delivery of biotherapeutics including anti-miRNA oligonucleotides (AMOs). This study is a continuation of earlier findings involving poly(ε-caprolactone) (PCL)-poly (ethylene glycol) (PEG) nanoparticles (PEG-PCL NPs), which were coated with trastuzumab to target GC with HER2 receptor over-expression using anti-miRNA-21 (AMO-21) and 5-Fu. Methods: HER-PEG-PCL NPs were prepared by one-step carbodiimide coupling using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride (EDAc) and Sulfo-NHS in aqueous phase. Covalent coupling of amino groups at the surface of PEG-PCL with the carboxyl groups of trastuzumab was analyzed by X-ray photoelectron spectroscopy (XPS). AMO-21/5-Fu NPs were formulated by a double-emulsion solvent evaporation technique. The cell line specificity, cellular uptake and AMO-21 delivery were investigated through the rhodamine-B-labeled 6-carboxyfluorescein (FAM)-AMO-21-PEG-PCL NPs coated with or without the antibody in both Her2-positive (NUGC4) and negative GC cells (SGC7901) visualized by fluorescence microscopy. The cytotoxicity of the HER-PEG-PCL NPs encapsulating AMO-21 was evaluated by MTT and apoptosis. Real-time reverse-transcription polymerase chain reaction (RT-PCR) was used to examine miR-21 and phosphatase and tensin homolog (PTEN) and Sprouty2 expression in GC cell lines. The antitumor effects of AMO-21/5-Fu NPs were compared with other groups in xenograft gastric cancer mice. Results: The antibody conjugates significantly enhanced the cellular uptake of NPs. The AMO-21/5-Fu NPs effectively suppressed the target miRNA expression in GC cells, which further up-regulated PTEN and Sprouty2. As a result, the sensitivity of HER2-expressing gastric cancer to trastuzumab and 5-Fu were enhanced both in vitro and in vivo. The approach enhanced the targeting by trastuzumab as well as antibody-dependent cellular cytotoxicity (ADCC) of immune effector cells Conclusions: Taken together, the results provide insight into the biological and clinical potential of targeted AMO-21 and 5-Fu co-delivery using modified trastuzumab for GC treatment

Effects of Concentration of Soybean Protein Isolate and Maltose and Oil Phase Volume Fraction on Freeze–Thaw Stability of Pickering Emulsion

There is growing interest in enhancing the freeze–thaw stability of a Pickering emulsion to obtain a better taste in the frozen food field. A Pickering emulsion was prepared using a two-step homogenization method with soybean protein and maltose as raw materials. The outcomes showed that the freeze–thaw stability of the Pickering emulsion increased when prepared with an increase in soybean protein isolate (SPI) and maltose concentration. After three freeze–thaw treatments at 35 mg/mL, the Turbiscan Stability Index (TSI) value of the emulsion was the lowest. At this concentration, the surface hydrophobicity (H0) of the composite particles was 33.6 and the interfacial tension was 44.34 mN/m. Furthermore, the rheological nature of the emulsions proved that the apparent viscosity and viscoelasticity of Pickering emulsions grew with a growing oil phase volume fraction and concentration. The maximum value was reached in the case of the oil phase volume fraction of 50% at a concentration of 35 mg/mL, the apparent viscosity was 18 Pa·s, the storage modulus of the emulsion was 575 Pa, and the loss modulus was 152 Pa. This research is significant for the production of freeze–thaw resistant products, and improvement of protein-stabilized emulsion products with high freeze–thaw stability