ZNF498 promotes hepatocellular carcinogenesis by suppressing p53-mediated apoptosis and ferroptosis via the attenuation of p53 Ser46 phosphorylation.

BACKGROUND: Dysfunctional p53 signaling is one of the major causes of hepatocellular carcinoma (HCC) tumorigenesis and development, but the mechanisms underlying p53 inactivation in HCC have not been fully clarified. The role of Krüppel-associated box (KRAB)-type zinc-finger protein ZNF498 in tumorigenesis and the underlying mechanisms are poorly understood. METHODS: Clinical HCC samples were used to assess the association of ZNF498 expression with clinicopathological characteristics and patient outcomes. A mouse model in which HCC was induced by diethylnitrosamine (DEN) was used to explore the role of ZNF498 in HCC initiation and progression. ZNF498 overexpression and knockdown HCC cell lines were employed to examine the effects of ZNF498 on cellular proliferation, apoptosis, ferroptosis and tumor growth. Western blotting, immunoprecipitation, qPCR, luciferase assays and flow cytometry were also conducted to determine the underlying mechanisms related to ZNF498 function. RESULTS: ZNF498 was found to be highly expressed in HCC, and increased ZNF498 expression was positively correlated with advanced pathological grade and poor survival in HCC patients. Furthermore, ZNF498 promoted DEN-induced hepatocarcinogenesis and progression in mice. Mechanistically, ZNF498 directly interacted with p53 and suppressed p53 transcriptional activation by inhibiting p53 Ser46 phosphorylation. ZNF498 competed with p53INP1 for p53 binding and suppressed PKCδ- and p53INP1-mediated p53 Ser46 phosphorylation. In addition, functional assays revealed that ZNF498 promoted liver cancer cell growth in vivo and in vitro in a p53-dependent manner. Moreover, ZNF498 inhibited p53-mediated apoptosis and ferroptosis by attenuating p53 Ser46 phosphorylation. CONCLUSIONS: Our results strongly suggest that ZNF498 suppresses apoptosis and ferroptosis by attenuating p53 Ser46 phosphorylation in hepatocellular carcinogenesis, revealing a novel ZNF498-PKCδ-p53INP1-p53 axis in HCC cells that would enrich the non-mutation p53-inactivating mechanisms in HCC.

Gammacerane suppresses the growth of human endometrial carcinoma cells via induction of apoptosis, cell cycle arrest, inhibition of cell migration, invasion and STAT3 signaling pathway.

PURPOSE: This study was conducted to assess the anticancer role of gammacerane against human endometrial cancer. METHODS: The human RL-95 cell line (endometrial cancer) and SV40 (normal endometrial cells) were used in this study. The MTT-based estimation of cell proliferation assay along with the colony formation assay were used for assessing the cell viability. Acridine orange (AO)/Ethidium bromide (EB) staining followed by fluorescent microscopy was performed for estimation of cell apoptosis. Flow cytometry was used to assess the cell cycle phase distribution of cancer cells. Cell migration and invasion were estimated using wound healing and transwell assay, respectively. Western blotting was used for protein expression studies. RESULTS: The cell proliferation assay revealed that gammacerane treatment led to loss of viability of RL-95 cancer cells in a concentration-dependent manner. However, the antiproliferative effects were comparatively less prominent when gammacerane was used against the SV40 normal endometrial cells. AO/EB staining of cancer cells showed that gammacerane is active in inducing apoptosis in RL-95 cells and apoptotic induction effects were more evident at higher concentrations of the molecule. Flow cytometric analysis with Annexin V-FITC/Propidium iodide (PI) fixed cells showed that the percentage of apoptotic cells increased with increase in gammacerane concentration. Apoptotic signal was mediated via the modulation of Bax/Bcl-2 protein ratio. Western blot analysis of STAT3 protein showed that gammacerane treatment reduced the protein levels of STAT3 and the effects were more prominent at higher treatment concentrations. CONCLUSION: Gammacerane, by its ability to take control over the transcription of STAT3 transcription factor, inhibits the proliferation of human endometrial cancer cells. The effects revealed loss of viability, arrest of mitosis and cellular apoptosis.

[Effects of Gui Zhi Fu Ling Jiao Nang on Hemorheology and Expression of Phenotypic Proteins in Uterine Smooth Muscle Cells in Rats with An Intrauterine Device].

Objective To observe the effect of Gui Zhi Fu Ling Jiao Nang (GZFLJN) on the expressions of alpha smooth muscle actin (α-SMA) and proliferating cell nuclear antigen (PCNA) in uterine vascular smooth muscle cells (VSMC) of rat models with an intrauterine device (IUD) and to determine the thromboxane B2 (TXB2) and 6-keto-prostaglandin F1α (6-keto-PGF1α) levels in peripheral blood. Methods Female Wistar rats were randomly divided into four groups:normal group (n=16,with normal breed without treatment),model group (n=18,drenching 0.9% normal saline after modeling of IUD),GZFLJN group (n=18),and aminocaproic acid tablets group (n=17). Immunohistochemical SP method was used to detect the expressions of α-SMA and PCNA in uterine VSMC.ELISA was served to detect the levels of TXB2 and 6-keto-PGF1α in peripheral blood. Results The positive rate of α-SMA were (50.89±9.41)%,(26.93±6.80)%,(48.92±6.80)%,and (34.63±7.26)%,respectively,in normal group,model group,GZFLJN group,and aminocaproic acid tablets group;obviously,it was significantly higher in normal group (t=14.43,P=0.00) and GZFLJN group (t=11.37,P=0.00) than that in model group and it was significantly lower in aminocaproic acid tablets group than in normal group (t=9.96,P=0.00) and GZFLJN group (t=8.23,P=0.00). The positive rate of PCNA were (25.66±7.24)%,(61.26±9.98)%,(28.36±9.17)%,and (50.23±8.71)%,respectively,in these four groups;obviously,it was significantly lower in the normal group (t=20.86,P=0.00) and GZFLJN group (t=19.12,P=0.00) than in model group and it was significantly higher in aminocaproic acid tablets group than in normal group (t=17.82,P=0.00) and GZFLJN group (t=16.05,P=0.00). Serum TXB2 level in these four groups were (445.86±24.43),(508.78±12.42),(448.11±9.63),and (498.11±13.63)ng/L;obviously,it was significantly higher in model group than in normal group (t=16.55,P=0.00) and aminocaproic acid tablets group (t=-4.12,P=0.00) and it was significantly lower in GZFLJN group than in model group (t=-15.23,P=0.00) and aminocaproic acid tablets group (t=-12.08,P=0.00). Serum 6-keto-PGF1α level in these four groups were (23.17±1.93),(18.09±0.93),(22.70±1.61),and (20.70±1.41)ng/L,respectively;obviously,it was significantly lower in model group than in normal group (t=-13.98,P=0.00) and aminocaproic acid tablets group (t=5.26,P=0.00) and it was significantly higher in GZFLJN group than in model group (t=11.43,P=0.00) and aminocaproic acid tablets group (t=8.76,P=0.00). Conclusion GZFLJN can regulate the expressions of α-SMA and PCNA of VSMC in the endometrium of IUD rats and the concentrations of TXB2 and 6-keto-PGF1α in the serum.

Surveillance of antimicrobial resistance among Escherichia coli from chicken and swine, China, 2008-2015.

The objective of this study was to investigate the antimicrobial resistance trend in Escherichia coli from food animals in China. During 2008-2015, a total of 15,130 E. coli were isolated from chicken and swine from seven provinces. The susceptibilities of these isolates to nine classes of antimicrobial agents were determined using broth microdilution susceptibility method. The findings of this study include: (1) multi-drug resistance was highly prevalent in E. coli; (2) these E. coli isolates showed high resistant rate (>80%) to several old drugs, including ampicillin, tetracycline and sulfisoxazole; (3) increasing resistance to colistin, florfenicol and ceftiofur was observed; (4) the E. coli isolates from different provinces had different resistance patterns. All these data highlight the rising problem of antimicrobial resistance. It is urgent to improve the management of animal production and enhance the proper use of antimicrobials in China as well as the other countries.

MiR-205 serves as a prognostic factor and suppresses proliferation and invasion by targeting insulin-like growth factor receptor 1 in human cervical cancer.

MicroRNAs are a kind of small and non-coding RNAs, which have been demonstrated to play an important role in the progression of human cervical cancer. Here, we found that the expression of miR-205 was low in cervical cancer cell lines and tissues, compared with matched non-tumor tissues and human endocervical epithelial cells. Also, miR-205 was inversely correlated with histological differentiation, metastasis, International Federation of Gynecology and Obstetrics stage, and the expression of insulin-like growth factor receptor 1 messenger RNA and protein. Besides, miR-205 or insulin-like growth factor receptor 1 expression is an independent prognostic factor. Mechanically, ectopic expression of miR-205 decreased proliferation, colony formation, and some proliferation/apoptosis-related proteins in cervical cancer cells. Ectopic expression of miR-205 caused G1 arrest. Luciferase reporter assays confirmed that binding of miR-205 to the 3' untranslated region of insulin-like growth factor receptor 1 may potentially decrease the expression of insulin-like growth factor receptor 1. Notably, insulin-like growth factor receptor 1 overexpression attenuated the inhibitory effects of miR-205 on cell proliferation and invasion, while small interfering RNA-insulin-like growth factor receptor 1 enhanced the inhibitory effects of miR-205 on cell proliferation and invasion. In conclusion, our findings suggested that miR-205 serves as a prognostic factor and suppresses proliferation and invasion by targeting insulin-like growth factor receptor 1 in human cervical cancer. Thus, miR-205/insulin-like growth factor receptor 1 pathway may be of great benefit to cervical cancer patients.

[Simultaneous determination of four fluoroquinolone residues in edible chicken tissues by reversed-phase high performance liquid chromatography].

A reversed-phase high performance liquid chromatographic method with fluorescence detection was developed for the simultaneous analysis of ciprofloxacin, danofloxacin, enrofloxacin and sarafloxacin residues in edible chicken tissues. The analytes were extracted from chicken muscle, skin and fat, liver, kidney by aqueous potassium dihydrogenphosphate of different pH values through homogenization. The supernatant of the extract was applied onto a C18 solid phase extraction cartridge for clean-up. The separation was achieved on a C18 column, and the detection was performed with a fluorescence detector (excitation at 280 nm and emission at 450 nm). The four fluoroquinolones were analyzed in spiked samples of four chicken tissues with mean recoveries in the range of 53.9%-93.4% at spiked levels of 20-300 microg/kg. The relative standard deviations of inter-assay were no more than 23%. The detection limits of quantification were 20 microg/kg for ciprofloxacin, enrofloxacin and sarafloxacin and 4 microg/kg for danofloxacin. The method is simple, fast, and sufficient for routine analysis.