Blockage of AMPK-ULK1 pathway mediated autophagy promotes cell apoptosis to increase doxorubicin sensitivity in breast cancer (BC) cells: an in vitro study.

BACKGROUND: Activation of autophagy flux contributed to resistance of breast cancer (BC) cells to current chemotherapeutic drugs, which seriously limited their therapeutic efficacy and facilitated BC recurrence in clinic. However, the detailed mechanisms are still not fully understood. In the present study, we identified that inactivation of AMPK-ULK1 signaling cascade mediated protective autophagy sensitized BC cells to doxorubicin in vitro. METHODS: Cell counting kit-8 (CCK-8) assay and colony formation assay were performed to evaluate cell proliferation abilities. Trypan blue staining assay was used to examine cell viability, and Annexin V-FITC/PI double staining method was conducted to determine cell apoptosis. The autophagosomes in BC cells were observed and photographed by electronic microscope (EM). Western Blot analysis was employed to examine genes expressions at protein levels. RESULTS: The parental doxorubicin-sensitive BC (DS-BC) cells were exposed to increasing concentrations of doxorubicin to establish doxorubicin-resistant BC (DR-BC) cells, and the DR-BC cells were much more resistant to high-dose doxorubicin treatment compared to the DS-BC cells. Interestingly, high-dose doxorubicin specifically increased LC3B-II/I ratio, promoted autophagosomes formation and decreased p62 expression levels to facilitate autophagy in DR-BC cells, instead of DS-BC cells, and the autophagy inhibitor 3-methyladenine (3-MA) enhanced the cytotoxic effects of high-dose doxorubicin on DR-BC cells. In addition, we proved that high-dose doxorubicin triggered protective autophagy in DR-BC cells by activating AMPK-ULK1 pathway. Functionally, high-dose doxorubicin increased the expression levels of phosphorylated AMPK (p-AMPK) and ULK1 (p-ULK1) to activate AMPK-ULK1 pathway in DR-BC cells, and the inhibitors for AMPK (compound C) and ULK1 (SBI-0206965) blocked autophagy to promote cell death and slow down cell growth in DR-BC cells treated with high-dose doxorubicin. CONCLUSIONS: Collectively, our in vitro data indicated that blockage of AMPK-ULK1 signaling cascade mediated protective autophagy might be a promising strategy to increase doxorubicin sensitivity for BC treatment.

New ß-Lactone with Tea Pathogenic Fungus Inhibitory Effect from Marine-Derived Fungus MCCC3A00957.

Fusarium solani H915 (MCCC3A00957), a fungus originating from mangrove sediment, showed potent inhibitory activity against tea pathogenic fungus Pestalotiopsis theae. Successive chromatographic separation on an ethyl acetate (EtOAc) extract of F. solani H915 resulted in the isolation of five new alkenoic diacid derivatives: fusarilactones A-C (1-3), and fusaridioic acids B (4) and C (5), in addition to seven known compounds (6-12). The chemical structures of these metabolites were elucidated on the basis of UV, IR, HR-ESI-MS, and NMR spectroscopic data. The antifungal activity of the isolated compounds was evaluated. Compounds with a ß-lactone ring (1, 2, and 7) exhibited potent inhibitory activities, while none of the other compounds show activity. The ED50 values of the compounds 1, 2, and 7 were 38.14 ± 1.67, 42.26 ± 1.96, and 18.35 ± 1.27 µg/mL, respectively. In addition, inhibitory activity of these compounds against 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase gene expression was also detected using real-time RT-PCR. Results indicated that compounds 1, 2, and 7 may inhibit the growth of P. theae by interfering with the biosynthesis of ergosterol by down-regulating the expression of HMG-CoA synthase.

A comparison of food crispness based on the cloud model.

The cloud model is a typical model which transforms the qualitative concept into the quantitative description. The cloud model has been used less extensively in texture studies before. The purpose of this study was to apply the cloud model in food crispness comparison. The acoustic signals of carrots, white radishes, potatoes, Fuji apples, and crystal pears were recorded during compression. And three time-domain signal characteristics were extracted, including sound intensity, maximum short-time frame energy, and waveform index. The three signal characteristics and the cloud model were used to compare the crispness of the samples mentioned above. The crispness based on the Ex value of the cloud model, in a descending order, was carrot > potato > white radish > Fuji apple > crystal pear. To verify the results of the acoustic signals, mechanical measurement and sensory evaluation were conducted. The results of the two verification experiments confirmed the feasibility of the cloud model. The microstructures of the five samples were also analyzed. The microstructure parameters were negatively related with crispness (p < .01). PRACTICAL APPLICATIONS: The cloud model method can be used for crispness comparison of different kinds of foods. The method is more accurate than the traditional methods such as mechanical measurement and sensory evaluation. The cloud model method can also be applied to other texture studies extensively.

[Influence of ablactation rats' high calcium intake to the expression of uncoupling protein 2 mRNA after adulthood obesity induction].

OBJECTIVE: To study the influence of early high calcium intake to adulthood obesity through detection the expression of uncoupling protein 2 ( UCP2) mRNA in muscle by a reverse transcription polymerase chain reaction ( RT-PCR) technique. METHODS: 120 male Wistar rats were divided randomly into normal control group, high dose calcium group, medium dose calcium group and low doses calcium group, with basic diet and high calcium diet for 4 weeks. After intervention, blood fat was compared. Subsequently, all rats were fed basic diet for 3 weeks and blood fat was compared. Then, normal feed rats were randomly divided into normal controls and obesity induction group. Obesity induction group, high dose calcium group, medium dose calcium group and low doses calcium group were fed high-fat food. After 8 weeks, RT-PCR was used for analysis After obesity induction, weight the expression of UCP2 mRNA in muscle. RESULTS: growth of three high calcium groups were below obesity induction group, weight growth of low doses and high doses of high calcium groups were no difference with normal control group. Serum triglyceride levels of high dose high calcium group were significantly lower than obesity induction group and no difference with normal control group. Expression level of UCP2 mRNA of obesity induction group and high doses of high calcium group were obviously lower than normal controls, low dose and medium dose high calcium group, medium dose high calcium group' expression was significantly higher than normal control group. CONCLUSION: In rats' early life high calcium intake can continue to affect adulthood obesity induced by high-fat feed, increase expression level of UCP2 mRNA, improve the disorder of blood fat metabolism.