Adhesion GPCR ADGRE2 Maintains Proteostasis to Promote Progression in Acute Myeloid Leukemia.

Acute myeloid leukemia (AML) is an aggressive and heterogeneous hematologic malignancy. In elderly patients, AML incidence is high and has a poor prognosis due to a lack of effective therapies. G protein-coupled receptors (GPCR) play integral roles in physiologic processes and human diseases. Particularly, one third of adhesion GPCRs, the second largest group of GPCRs, are highly expressed in hematopoietic stem and progenitor cells or lineage cells. Here, we investigate the role of adhesion GPCRs in AML and whether they could be harnessed as antileukemia targets. Systematic screening of the impact of adhesion GPCRs on AML functionality by bioinformatic and functional analyses revealed high expression of ADGRE2 in AML, particularly in leukemic stem cells, which is associated with poor patient outcomes. Silencing ADGRE2 not only exerts antileukemic effects in AML cell lines and cells derived from patients with AML in vitro, but also delays AML progression in xenograft models in vivo. Mechanistically, ADGRE2 activates phospholipase Cß/protein kinase C/MEK/ERK signaling to enhance the expression of AP1 and transcriptionally drive the expression of DUSP1, a protein phosphatase. DUSP1 dephosphorylates Ser16 in the J-domain of the co-chaperone DNAJB1, which facilitates the DNAJB1-HSP70 interaction and maintenance of proteostasis in AML. Finally, combined inhibition of MEK, AP1, and DUSP1 exhibits robust therapeutic efficacy in AML xenograft mouse models. Collectively, this study deciphers the roles and mechanisms of ADGRE2 in AML and provides a promising therapeutic strategy for treating AML. Significance: Increased expression of the adhesion GPCR member ADGRE2 in AML supports leukemia stem cell self-renewal and leukemogenesis by modulating proteostasis via an MEK/AP1/DUSP1 axis, which can be targeted to suppress AML progression.

Mapping the EORTC QLQ-C30 and QLQ H&N35 to the EQ-5D-5L and SF-6D for papillary thyroid carcinoma.

PURPOSE: Empirical evidence for the EORTC QLQ C30 scale in thyroid cancer mapping algorithms has not been found in China, which limits the cost-utility analysis of patients with papillary thyroid carcinoma (PTC) population. We developed mapping algorithms that use the EORTC QLQ-C30 and QLQ H&N35 to predict EQ-5D-5L and SF-6D health utility scores for PTC patients. METHODS: Data from 1050 Chinese PTC patients who completed the EORTC QLQ-C30, QLQ H&N35, EQ-5D-5L and SF-6D instruments were collected. Direct mapping (OLS, Tobit, Betamix) and indirect mapping functions (Order Probit) were used to estimate algorithms. The goodness-of-fit of mapping performance was assessed by MAE, RMSE, AIC, BIC, AE, and ICC. A fivefold cross-validation and random sample validation approach were used to test the stability of the models. RESULTS: The mean EQ-5D-5L and SF-6D utility scores were 0.8704 and 0.6368, respectively. We recommend the Betamix model for the EQ-5D-5L (MAE = 0.0363, RMSE = 0.0505, AIC = -3458.73, BIC = -3096.91, AE > 0.05(%) = 48.38, AE > 0.1(%) = 8.67, ICC = 0.8288 for the full sample dataset) and the Betamix model for the SF-6D (MAE = 0.0328, RMSE = 0.0417, AIC = -2788.91, BIC = -2605.51, AE > 0.05(%) = 42.76, AE > 0.1(%) = 3.62, ICC = 0.8657 for the full sample dataset), with EORTC QLQ-C30 all items, QLQ H&N35 all items, age and gender as the predicted variables showing the best performance. CONCLUSION: In the absence of preference-based quality of life tools, the mapping algorithms reported here are effective alternative for predicting the health utility of PTC patients, contributing to the cost-utility analysis studies.

Development of algorithms to estimate the EQ-5D-5L from the FACT-L in patients with lung cancer: a mapping study.

OBJECTIVE: This study aimed to develop a mapping algorithm to evaluate the EQ-5D-5L according to the FACT-L when the EQ-5D-5L is not available. METHODS: EQ-5D-5L and FACT-L data were collected from patients with lung cancer in Departments of Thoracic Surgery, Medical Oncology, Radiation Oncology, Sichuan Cancer Hospital. We used the ordinary least squares model (OLS), Tobit model (Tobit), two-part model (TPM), beta mixture regression (BM), and censored least absolute deviation model (CLAD) to map the results of the FACT-L according to EQ-5D-5L scores. To establish these models, the total score, dimension scores, squared items, and interaction items were introduced. Performance metrics including Adjusted R2, root mean square error (RMSE), and mean absolute error (MAE) were used to select the optimized model. RESULTS: The model with the best mapping performance was the BM model (BETAMIX4) with the PWB (physical well-being) dimension, FWB (functional well-being) dimension, the squared term of the PWB dimension, and the squared term of the FWB dimension as covariates. The final prediction metrics were Adjusted R2 = 0.695, RMSE = 0.206, and MAE = 0.109. Fivefold cross-validation (CV) results also demonstrated that the BM model had the best mapping power. CONCLUSIONS: This study developed an optimized mapping algorithm to predict the utility index from the FACT-L to the EQ-5D-5L, which provides an effective alternative reference for EQ-5D-5L estimation when the preference-based health utility values were unavailable.

Prediction of the SF-6D utility score from Lung cancer FACT-L: a mapping study in China.

OBJECTIVE: To develop a mapping algorithm for generating the Short Form Six-Dimension (SF-6D) utility score based on the Functional Assessment of Cancer Therapy-Lung (FACT-L) of lung cancer patients. METHODS: Data were collected from 625 lung cancer patients in mainland China. The Spearman rank correlation coefficient and principal component analysis were used to evaluate the conceptual overlap between the FACT-L and SF-6D. Five model specifications and four statistical techniques were used to derive mapping algorithms, including ordinary least squares (OLS), Tobit and beta-mixture regression models, which were used to directly estimate health utility, and ordered probit regression was used to predict the response level. The prediction performance was evaluated using the correlations between the root mean square error (RMSE), mean absolute error (MAE), concordance correlation coefficient (CCC), Akaike information criterion (AIC) and Bayesian information criterion (BIC) and the observed and predicted SF-6D scores. A five-fold cross-validation method was used to test the universality of each model and select the best model. RESULTS: The average FACT-L score was 103.024. The average SF-6D score was 0.774. A strong correlation was found between FACT-L and SF-6D scores (ρ = 0.797). The ordered probit regression model with the total score of each dimension and its square term, as well as age and sex as covariates, was most suitable for mapping FACT-L to SF-6D scores (5-fold cross-validation: RMSE = 0.0854; MAE = 0.0655; CCC = 0.8197; AEs > 0.1 (%) = 53.44; AEs > 0.05 (%) = 21.76), followed by beta-mixture regression for direct mapping. The Bland‒Altman plots showed that the ordered probit regression M5 had the lowest proportion of prediction scores outside the 95% agreement limit (-0.166, 0.163) at 4.96%. CONCLUSIONS: The algorithm reported in this paper enables lung cancer data from the FACT-L to be mapped to the utility of the SF-6D. The algorithm allows the calculation of quality-adjusted life years for cost-utility analyses of lung cancer.

Obtaining SF-6D utilities from FACT-H&N in thyroid carcinoma patients: development and results from a mapping study.

Objective: There is limited evidence for mapping clinical tools to preference-based generic tools in the Chinese thyroid cancer patient population. The current study aims to map the FACT-H&N (Functional Assessment of Cancer Therapy-Head and Neck Cancer) to the SF-6D (Short Form Six-Dimension), which will inform future cost-utility analyses related to thyroid cancer treatment. Methods: A total of 1050 participants who completed the FACT-H&N and SF-6D questionnaires were included in the analysis. Four methods of direct and indirect mapping were estimated: OLS regression, Tobit regression, ordered probit regression, and beta mixture regression. We evaluated the predictive performance in terms of root mean square error (RMSE), mean absolute error (MAE), concordance correlation coefficient (CCC), Akaike information criterion (AIC) and Bayesian information criterion (BIC) and the correlation between the observed and predicted SF-6D scores. Results: The mean value of SF-6D was 0.690 (SD = 0.128). The RMSE values for the fivefold cross-validation as well as the 30% random sample validation for multiple models in this study were 0.0833-0.0909, MAE values were 0.0676-0.0782, and CCC values were 0.6940-0.7161. SF-6D utility scores were best predicted by a regression model consisting of the total score of each dimension of the FACT-H&N, the square of the total score of each dimension, and covariates including age and gender. We proposed to use direct mapping (OLS regression) and indirect mapping (ordered probit regression) to establish a mapping model of FACT-H&N to SF-6D. The mean SF-6D and cumulative distribution functions simulated from the recommended mapping algorithm generally matched the observed ones. Conclusions: In the absence of preference-based quality of life tools, obtaining the health status utility of thyroid cancer patients from directly mapped OLS regression and indirectly mapped ordered probit regression is an effective alternative.

Pancreatic neuroendocrine tumor detected by technetium-99m methoxy-2-isobutylisonitrile single photon emission computed tomography/computed tomography: A case report.

BACKGROUND: Pancreatic neuroendocrine tumors (NETs) account for about 1%-2% of pancreatic tumors and about 8% of all NETs. Computed tomography (CT), magnetic resonance imaging, and endoscopic ultrasound are common imaging modalities for the diagnosis of pancreatic NETs. Furthermore, somatostatin receptor imaging is of great value for diagnosing pancreatic NETs. Herein, we report the efficacy of technetium-99m methoxy-2-isobutylisonitrile (99mTc-MIBI) single photon emission CT (SPECT)/CT for detecting pancreatic NETs. CASE SUMMARY: A 57-year-old woman presented to our hospital with a 1-d history of persistent upper abdominal distending pain. The distending pain in the upper abdomen was aggravated after eating, with nausea and retching. Routine blood test results showed a high neutrophil percentage, low leukomonocyte and monocyte percentages, and low leukomonocyte and eosinophil counts. Amylase, liver and kidney function, and tumor markers alpha-fetoprotein, carcinoembryonic antigen, and cancer antigen (CA) 125, CA72-4, CA19-9, and CA153 were normal. Abdominal CT showed a mass, with multiple calcifications between the pancreas and the spleen. The boundary between the mass and the pancreas and spleen was poorly defined. Contrast-enhanced CT revealed that the upper abdominal mass was unevenly and gradually enhanced. 99mTc-MIBI SPECT/CT revealed that a focal radioactive concentration, with mild radioactive concentration extending into the upper abdominal mass, was present at the pancreatic body and tail. The 99mTc-MIBI SPECT/CT manifestations were consistent with the final pathological diagnosis of pancreatic NET. CONCLUSION: 99mTc-MIBI SPECT/CT appears to be a valuable tool for detecting pancreatic NETs.

Mapping study of papillary thyroid carcinoma in China: Predicting EQ-5D-5L utility values from FACT-H&N.

Objective: To develop a mapping algorithm that can be used to predict EQ-5D-5L health utility scores from FACT-H&N and obtain health utility parameters for Chinese patients with papillary thyroid carcinoma (PTC), which can be used for cost-utility analysis in health economic. Methods: A total of 1,050 patients with PTC from a tertiary hospital in China were included, and they completed FACT-H&N and EQ-5D-5L. Four mapping algorithms of direct mapping functions were used to derive the models: Ordinary least squares (OLS), Tobit model (Tobit), Two-part model (TPM), and Beta mixture regression model (Beta). The goodness-of-fit of models was assessed by the mean absolute error (MAE), root mean square error (RMSE), Akaike information criteria (AIC), Bayesian information criteria (BIC), and absolute error (AE). A fivefold cross-validation method was used to test the stability of the models. Results: The mean utility value of the EQ-5D-5L was 0.870 ± 0.094. The mean EQ-VAS score was 76.5 ± 13.0. The Beta mixture regression model mapping FACT-H&N to EQ-5D-5L achieved the best performance [fivefold cross-validation MAE = 0.04612, RMSE = 0.06829, AIC = -2480.538, BIC = -2381.137, AE > 0.05 (%) = 32.48, AE > 0.1 (%) = 8.95]. The independent variables in this model were Physical Well-Being (PWB), Emotional Well-Being (EWB), Head & Neck Cancer Subscale (HNCS) scores and its square term and interaction term scores. Conclusions: This study calculated the health utility score of Chinese patients with PTC. The reported algorithms can be used to map the FACT-H&N into the EQ-5D-5L, which can be applied in the cost-utility related study of patients with PTC.

NRF-2α and mitophagy underlie enhanced mitochondrial functions and biogenesis induced by T-2 toxin in GH3 cells.

T-2 toxin is a natural contaminant in grain cereals produced by species of Fusarium. Studies indicate that T-2 toxin can positively affect mitochondrial function, but the underlying mechanism is unclear. In this study, we examined the role of nuclear respiratory factor 2α (NRF-2α) in T-2 toxin-activated mitochondrial biogenesis and the direct target genes of NRF-2α. Furthermore, we investigated T-2 toxin-induced autophagy and mitophagy, and the role of mitophagy in changes in mitochondrial function and apoptosis. It was found that T-2 toxin significantly increased NRF-2α levels and nuclear localization of NRF-2α was induced. NRF-2α deletion significantly increased the production of reactive oxygen species (ROS), abrogated T-2 toxin-induced increases in ATP and mitochondrial complex I activity, and inhibited the mitochondrial DNA copy number. Meanwhile, With chromatin immunoprecipitation sequencing (ChIP-Seq), various novel NRF-2α target genes were identified, such as mitochondrial iron-sulphur subunits (Ndufs 3,7) and mitochondrial transcription factors (Tfam, Tfb1m, and Tfb2m). Some target genes were also involved in mitochondrial fusion and fission (Drp1), mitochondrial translation (Yars2) and splicing (Ddx55), and mitophagy. Further studies showed that T-2 toxin induced Atg5 dependent autophagy and Atg5/PINK1-dependent mitophagy. In addition, mitophagy defects increase ROS production, inhibit ATP levels and the expression of genes related to mitochondrial dynamics, and promote apoptosis in the presence of T-2 toxins. Altogether, these results suggest that NRF-2α plays a critical role in promoting mitochondrial function and biogenesis through regulation of mitochondrial genes, and, interestingly, mitophagy caused by T-2 toxin positively affected mitochondrial function and protected cell survival against T-2 toxin.

Dlk1 maintains adult mice long-term HSCs by activating Notch signaling to restrict mitochondrial metabolism.

BACKGROUND: Adult hematopoietic stem cells (HSCs) homeostasis is critically important in maintaining lifelong hematopoiesis. However, how adult HSCs orchestrate its homeostasis remains not fully understood. Imprinted gene Dlk1 has been shown to play critical role in mouse embryonic hematopoiesis and in regulation of stem cells, but its physiological roles in adult HSCs are unknown. METHODS: We performed gene expression analysis of Dlk1, and constructed conditional Dlk1 knockout (KO) mice by crossing Mx1 cre mice with Dlkflox/flox mice. Western blot and quantitative PCR were used to detect Dlk1 KO efficiency. Flow cytometry was performed to investigate the effects of Dlk1 KO on HSCs, progenitors and linage cells in primary mice. Competitive HSCs transplantation and secondary transplantation was used to examine the effects of Dlk1 KO on long-term hematopoietic repopulation potential of HSCs. RNA-Seq and cell metabolism assays was used to determine the underlying mechanisms. RESULTS: Dlk1 was highly expressed in adult mice long-term HSCs (LT-HSCs) relative to progenitors and mature lineage cells. Dlk1 KO in adult mice HSCs drove HSCs enter active cell cycle, and expanded phenotypical LT-HSCs, but undermined its long-term hematopoietic repopulation potential. Dlk1 KO resulted in an increase in HSCs' metabolic activity, including glucose uptake, ribosomal translation, mitochondrial metabolism and ROS production, which impaired HSCs function. Further, Dlk1 KO in adult mice HSCs attenuated Notch signaling, and re-activation of Notch signaling under Dlk1 KO decreased the mitochondrial activity and ROS production, and rescued the changes in frequency and absolute number of HSCs. Scavenging ROS by antioxidant N-acetylcysteine could inhibit mitochondrial metabolic activity, and rescue the changes in HSCs caused by Dlk1 KO. CONCLUSION: Our study showed that Dlk1 played an essential role in maintaining HSC homeostasis, which is realized by governing cell cycle and restricting mitochondrial metabolic activity.

Adipose-derived mesenchymal stem cell-secreted extracellular vesicles alleviate non-alcoholic fatty liver disease via delivering miR-223-3p.

Increasing studies have identified the potential of mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) in non-alcoholic fatty liver disease (NAFLD) treatment. Hence, we further focused on the potential of adipose-derived MSC (ADSC)-EVs in NAFLD by delivering miR-223-3p. The uptake of isolated ADSC-EVs by hepatocytes was assessed, and the expression of miR-223-3p in ADSC-EVs and hepatocytes was characterized. It was established that miR-223-3p, enriched in ADSC-EVs, could be delivered by ADSC-EVs into hepatocytes. Using co-culture system and gain-of-function approach, we evaluated the effect of ADSC-EVs carrying miR-223-3p on lipid accumulation and liver fibrosis in pyrrolizidine alkaloids (PA)-induced hepatocytes and a high-fat diet-induced NAFLD mouse model. Bioinformatics websites and dual-luciferase reporter gene assay were performed to determine the interactions between miR-223-3p and E2F1, which was further validated by rescue experiments. ADSC-EVs containing miR-223-3p displayed suppressive effects on lipid accumulation and liver fibrosis through E2F1 inhibition, since E2F1 was demonstrated as a target gene of miR-223-3p. The protective role of ADSC-EVs by delivering miR-223-3p was then confirmed in the mouse model. Collectively, this study elucidated that ADSC-EVs delayed the progression NAFLD through the delivery of anti-fibrotic miR-223-3p and subsequent E2F1 suppression, which may suggest miR-223-3p-loaded ADSC-EVs to be a potential therapeutic approach for NAFLD.

Molecular and epidemiological characterization of human adenoviruses infection among children with acute diarrhea in Shandong Province, China.

BACKGROUND: Human adenovirus (HAdV) had been recognized as one of the most common enteric viruses associated with acute diarrhea in children. The present study was carried out to demonstrate the molecular and epidemiological characterization of HAdV Infections among children in Shandong province in China between July 2017 and June 2018. METHODS: Fecal specimens were collected from children under 5 years old with acute diarrhea. DNA was extracted from the stool specimens and adenovirus DNA was detected by PCR amplification with specific primers. The amplification products were subjected to electrophoresis and visualized on a UV transilluminator. All positive RT-PCR amplification products were sequenced and the obtained sequences analyzed by MEGA (version 7.0). Demographic information and clinical manifestation data were also analyzed. RESULTS: In total, 656 fecal specimens were collected and the overall positive rate of HAdV was 7.47%. HAdV infections were detected in all age groups, in which children aged 13-24 months presented the highest positive rate. Seasonal pattern could be observed with a peak in December, January and February. Diarrhea, vomiting, dehydration and fever were the main clinical manifestations, in which vomiting was the most common accompanied symptom. By phylogenetic analysis, four species (A, B, C, and F) were detected and seven different serotypes were identified. HAdV-41 (48.98%, 24/49) was the most common serotype followed by HAdV-3 (18.37%, 9/49), HAdV-31 (14.29%, 7/49), HAdV-7 (8.16%, 4/49), HAdV-40 (4.08%, 2/49), HAdV-1 (4.08%, 2/49) and HAdV-2 (2.04%, 1/49). CONCLUSION: This study indicated that HAdV infection was an important cause of acute diarrhea among children under 5 years old in Shandong province. The results will contribute to (a) increase understanding of the role of HAdV in diarrheal children and enhance identification of the predominant diarrhea pathogen for diagnosis; (b) avoid abuse of antibiotics; (c) monitor the change of prevalent HAdV serotypes and promote vaccine development and vaccination.

[Acupoint catgut embedding improves gastric mucosal injury by down-regulating expression of HIF-1α and VEGF in chronic atrophic gastritis rats].

OBJECTIVE: To observe the effect of acupoint catgut embedding on histopathological changes of gastric mucosa and expression of mucosal hypoxia inducible factor (HIF)-1α and vascular endothelial growth factor (VEGF) proteins in chronic atrophic gastritis (CAG) rats, so as to explore its mechanisms underlying improvement of CAG. METHODS: Male SD rats were divided into blank group (n=10) and model group (n=9) and catgut embedment group (n=10). The CAG model was established by free drinking of N-methyl-N'-nitro-N-nitroso-guanidine solution (100 µg/mL) and irregular diet. Catgut embedment was applied to "Zhongwan" (CV12) and bilateral "Zusanli" (ST36) and "Pishu" (BL20), once every 10 days, 6 times in total. Histopathological changes of gastric mucosal tissue were observed under microscope after H.E. staining. The expression of HIF-1α and VEGF proteins in the antrum of stomach was detected by Western blot. RESULTS: H.E. staining showed that compared with the blank group, the number of rats with glandular necrosis, atrophy and mucosal stasis in the model group were increased significantly (P<0.001, P<0.05).Compared with the model group, the number of rats with gland necrosis, atrophy and mucosal stasis in the catgut embedment group were significantly reduced (P<0.01, P<0.05). Western blot displayed that the expression levels of HIF-1α and VEGF proteins were significantly higher in rats with blood stasis and model group than in those without blood stasis and the blank group (P<0.01), and were considerably down-regulated in the catgut embedment group than in the model group (P<0.05, P<0.01). CONCLUSION: Acupoint catgut embedment can improve the injury of gastric mucosa in CAG rats, which may be associated with its function in down-regulating the expression of HIF-1α and VEGF proteins in the gastric mucosa.

The epigenetic mechanisms in Fusarium mycotoxins induced toxicities.

Fusarium mycotoxins are the most economically important fungal toxins. Fumonisins, zearalenone and trichothecenes (T-2 toxin, HT-2 toxin, deoxynivalenol, nivalenol etc) are the major representatives and most studied of Fusarium mycotoxins. The Fusarium mycotoxins contaminate cereal grains, animal feeds and human food products, and cause huge economic losses and pose a threat to animal and human health globally. Depending on the type, the toxicity of Fusarium mycotoxins and the mechanisms have been well investigated. Epigenetic modifications (DNA methylation, histone modifications and regulation of non-coding RNA) have been implicated in various human diseases and the toxicities in animals caused by Fusarium mycotoxins, including carcinogenesis, genotoxicity and reproductive disorders. On the basis of recently documented data, this review discussed the relationship between the epigenetic modifications and Fusarium mycotoxins-induced toxicities.

An overview of epigenetic agents and natural nutrition products targeting DNA methyltransferase, histone deacetylases and microRNAs.

Several humans' diseases such as; cancer, heart disease, diabetes retain an etiology of epigenetic, and a new therapeutic option termed as "epigenetic therapy" can offer a potential way to treat these diseases. A numbers of epigenetic agents such as; inhibitors of DNA methyltransferase (DNMT) and histone deacetylases (HDACs) have grew an intensive investigation, and many of these agents are currently being tested in a clinical trial, while some of them have been approved for the use by the authorities. Since miRNAs can act as tumor suppressors or oncogenes, the miRNA mimics and molecules targeted at miRNAs (antimiRs) have been designed to treat some of the diseases. Much naturally occurring nutrition were discovered to alter the epigenetic states of cells. The nutrition, including polyphenol, flavonoid compounds, and cruciferous vegetables possess multiple beneficial effects, and some can simultaneously change the DNA methylation, histone modifications and expression of microRNA (miRNA). This review mainly summarizes the information of epigenetic agents of DNMTs and HDACs inhibitors, miRNA mimics and antimiRs, as well as the natural nutrition. In addition, some future perspectives related to the epigenetic therapy are also included.

Corrigendum: Mequindox Induced Genotoxicity and Carcinogenicity in Mice.

[This corrects the article DOI: 10.3389/fphar.2018.00361.].

Signaling pathways involved in the expression of SZNF and the target genes binding with SZNF related to cyadox.

SZNF (Sus scrofa zinc finger CCHC domain containing 3) is a post-transcription regulation factor, belonging to CCHC zinc finger proteins. Cyadox is a novel quinoxaline drug with antibacterial and growth promotion effects. In this study, we investigated the pharmacological mechanism of cyadox mediated by SZNF. Firstly, signaling pathways related to cyadox-induced SZNF expression were studied. The results showed that the mRNA level of SZNF reached the peak as early as 4 h after 2 µM cyadox treatment in swine hepatocytes. Several signaling pathways, including JAK2/STAT1, PI3K/Akt, TGF-ß/Smad3 and p38, might play critical roles in regulation of SZNF. The JAK2/STAT1, JAK2/PI3K/Akt, PI3K/Akt and myD88 & TAK1 & ASK1 /P38 signaling pathways were firstly activated after cyadox treatment in swine hepatocyte, the TGF-ß/Smad3 signaling pathway was activated later. Then given the characteristic of RNA binding of CCHC zinc finger proteins, the target mRNAs binding with SZNF were detected by RNA immunoprecipitation coupled to sequencing (RIP-seq) in PK-15 cells treated with cyadox. The RIP-Seq results showed that the bound mRNAs of 45 genes and 93 genes by SZNF protein were increased and decreased, respectively in cyadox-treated PK-15 cells compared with blank sample. With bioinformatics analysis, we showed that cyadox might exert its antibacterial and growth promotion effect by regulating SZNF-associated target genes in post-transcriptional level, such as genes related to growth (MLXIP, CKS2) and inflammation (LGALS3, PLAU). Thus, our results indicated that SZNF can post-transcriptionally regulate its target genes related to growth and inflammatory in cyadox-treated cells, which may explain the pharmacological mechanism of this drug.

Knockdown long non-coding RNA ANRIL inhibits proliferation, migration and invasion of HepG2 cells by down-regulation of miR-191.

BACKGROUND: Hepatocellular carcinoma (HCC) is a common malignant tumor with high fatality rate. Recent studies reported that up-regulation of long non-coding RNA antisense non-coding RNA in the INK4 locus (lncRNA ANRIL) was found in HCC tissues, and which could affect HCC cells biological processes. However, the potential molecular mechanism of ANRIL in HCC is still unclear. The study aimed to uncover the effect of ANRIL on HepG2 cells growth, migration and invasion. METHODS: The knockdown expression vectors of ANRIL were transfected into HepG2 cells, and qRT-PCR, CCK-8, flow cytometry, Transwell and western blot assays were performed to analyze the effect of ANRIL on cell proliferation, apoptosis, migration and invasion. The relative expression of miR-191 was then examined in ANRIL knockdown vector transfected cells. These experiments were repeated again for exploring the effect of miR-191 on HepG2 cells. NF-κB and Wnt/ß-catenin signaling pathways were examined by using western blot assay. RESULTS: Knockdown of ANRIL inhibited proliferation, induced apoptosis, meanwhile suppressed migration and invasion of HepG2 cells. Additionally, the results showed that the expression level of miR-191 was down-regulated by ANRIL knockdown in HepG2 cells. Importantly, overexpression of miR-191 reversed the anti-tumor effect of ANRIL on cell proliferation, apoptosis, migration and invasion in HepG2 cells. Besides, we found that ANRIL knockdown inactivated NF-κB and Wnt/ß-catenin pathways by regulating miR-191. CONCLUSIONS: These data demonstrated that ANRIL knockdown suppressed proliferation, migration, invasion, and promoted apoptosis in HepG2 cells by down-regulating miR-191 and inactivating NF-κB and Wnt/ß-catenin signaling pathways.

Mequindox Induced Genotoxicity and Carcinogenicity in Mice.

Mequindox (MEQ), acting as an inhibitor of deoxyribonucleic acid (DNA) synthesis, is a synthetic heterocyclic N-oxides. To investigate the potential carcinogenicity of MEQ, four groups of Kun-Ming (KM) mice (50 mice/sex/group) were fed with diets containing MEQ (0, 25, 55, and 110 mg/kg) for one and a half years. The result showed adverse effects on body weights, feed consumption, hematology, serum chemistry, organ weights, relative organ weights, and incidence of tumors during most of the study period. Treatment-related changes in hematology, serum chemistry, relative weights and histopathological examinations revealed that the hematological system, liver, kidneys, and adrenal glands, as well as the developmental and reproductive system, were the main targets after MEQ administration. Additionally, MEQ significantly increased the frequency of micronucleated normochromatic erythrocytes in bone marrow cells of mice. Furthermore, MEQ increased the incidence of tumors, including mammary fibroadenoma, breast cancer, corticosuprarenaloma, haemangiomas, hepatocarcinoma, and pulmonary adenoma. Interestingly, the higher incidence of tumors was noted in M25 mg/kg group, the lowest dietary concentration tested, which was equivalent to approximately 2.25 and 1.72 mg/kg b.w./day in females and males, respectively. It was assumed that the lower toxicity might be a reason for its higher tumor incidence in M25 mg/kg group. This finding suggests a potential relationships among the dose, general toxicity and carcinogenicity in vivo, and further study is required to reveal this relationship. In conclusion, the present study demonstrates that MEQ is a genotoxic carcinogen in KM mice.