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Background: HMGA2 encodes a small non histone chromatin-associated protein that has no intrinsic transcriptional activity, but can modulate transcription by altering the chromatin architecture. HMGA2 was found overexpressed in a variety of epithelial and mesenchymal tumors and promoted invasion and metastasis in most malignant epithelial tumors. A recent study showed that P53 inhibited CRC progression by targeting HMGA2. However, the mechanism by which HMGA2 affect angiogenesis in CRC has not been clarified. Methods: The expression of HMGA2 was analyzed by IHC, WB and bio infomatic analysis. Cbioportal and mexpress online tools were applied to explore the CNV and methylation of HMGA2 in CRC patients. Single cell data from GEO was used to examine the specific cell type that contribute to the high HMGA2 expression in CRC. Lentivirus was used to knock down HMGA2 in CRC cells and HUVECs was used to study angiogenesis. Results: In the current study, we first detected the expression pattern of HMGA2 in CRC patients and evaluated its clinical values and CNV amplification could possibly contribute to the up regulation of HMGA2 in CRC patients. By analyzing CRC single cell data we found that HMGA2 was specifically up regulated in the colorectal epithelial cells. Furthermore, knocking down of HMGA2 suppresses angiogenesis via dual regulation of VEGF-A and SEMA3A in CRC through inactivating VEGRR2 pathway in HUVECs. Conclusions: HMGA2 might be a promising prognostic marker and target for treating advanced CRC patients.

Summary Inverted feeding is a paradigm to study synchronization of circadian clocks by feeding rhythm in tissues more directly. Here, we provide a protocol for performing inverted feeding in mice and analyzing circadian rhythmicity in mouse tissues. We describe setting up inverted feeding and performing tissue dissection, followed by RNA extraction and gene expression analysis, and lastly R software-based analysis of circadian rhythmicity. This protocol can be combined with the use of CircaMetDB database for mechanistic studies of inverted feeding. For complete details on the use and execution of this protocol, please refer to Xin et al. (2021)., Graphical abstract, Highlights • Set up inverted feeding (IF) to evaluate clock synchronization by meal time • Hands-on tutorial of circadian rhythmicity detection and comparison in R • Protocol combined with CircaMetDB resource can be used for mechanistic study of IF, Inverted feeding is a paradigm to study synchronization of circadian clocks by feeding rhythm in tissues more directly. Here, we provide a protocol for performing inverted feeding in mice and analyzing circadian rhythmicity in mouse tissues. We describe setting up inverted feeding and performing tissue dissection, followed by RNA extraction and gene expression analysis, and lastly R software-based analysis of circadian rhythmicity. This protocol can be combined with the use of CircaMetDB database for mechanistic studies of inverted feeding.

Dysregulation of long noncoding RNA (lncRNA) is known to be involved in numerous human diseases, including lung cancer. However, the precise biological functions of most lncRNA remain to be elucidated. Here, we identified a novel up-regulated lncRNA, LINC01436 (RefSeq: NR_110419.1), in non-small cell lung cancer (NSCLC). High expression of LINC01436 was significantly associated with poor overall survival. Notably, LINC01436 expression was transcriptionally repressed by E2F6 under normoxia, and the inhibitory effect was relieved in a hypoxic microenvironment. Gain- and loss-of-function studies revealed that LINC01436 acted as a proto-oncogene by promoting lung cancer cell growth, migration and invasion in vitro. Xenograft tumor assays in nude mice confirmed that LINC01436 promoted tumor growth and metastasis in vivo. Mechanistically, LINC01436 exerted biological functions by acting as a microRNA (miR)-30a-3p sponge to regulate the expression of its target gene EPAS1. Our findings characterize LINC01436 as a new hypoxia-sensitive lncRNA with oncogenic function in NSCLC, suggesting that LINC01436 may be a potential biomarker for prognosis and a potential target for treatment.

Summary Time of eating synchronizes circadian rhythms of metabolism and physiology. Inverted feeding can uncouple peripheral circadian clocks from the central clock located in the suprachiasmatic nucleus. However, system-wide changes of circadian metabolism and physiology entrained to inverted feeding in peripheral tissues remain largely unexplored. Here, we performed a 24-h global profiling of transcripts and metabolites in mouse peripheral tissues to study the transition kinetics during inverted feeding, and revealed distinct kinetics in phase entrainment of diurnal transcriptomes by inverted feeding, which graded from fat tissue (near-completely entrained), liver, kidney, to heart. Phase kinetics of tissue clocks tracked with those of transcriptomes and were gated by light-related cues. Integrated analysis of transcripts and metabolites demonstrated that fatty acid oxidation entrained completely to inverted feeding in heart despite the slow kinetics/resistance of the heart clock to entrainment by feeding. This multi-omics resource defines circadian signatures of inverted feeding in peripheral tissues (www.CircaMetDB.org.cn)., Graphical abstract, Highlights • A multi-omics analysis of food entrainment in mouse peripheral tissues • Inverted feeding rhythm entrains diurnal transcriptomes with distinct kinetics • Phase kinetics of tissue clocks is conditioned by constant light • Cardiac metabolism entrains to feeding fast with slow kinetics of the heart clock, Animal Physiology ; Systems Biology ; Metabolomics ; Transcriptomics

Time of eating synchronizes circadian rhythms of metabolism and physiology. Inverted feeding reverses peripheral circadian clocks, but privileges the light-responsive central clock located in the suprachiasmatic nucleus. However, system-wide changes of circadian metabolism and physiology entrained to inverted feeding in peripheral tissues remain largely unexplored. Here, we performed a 24-h global profiling of transcripts and metabolites in mouse peripheral tissues to study the transition kinetics during inverted feeding, and revealed distinct kinetics in phase entrainment of diurnal transcriptomes by inverted feeding, which graded from fat tissue (near-completely entrained), liver, kidney, to heart. Phase kinetics of tissue clocks tracked with those of transcriptomes, and was gated by light cue. Integrated analysis of transcripts and metabolites demonstrated that fatty acid oxidation entrained completely to inverted feeding in heart despite the slow kinetics of the heart clock. This multi-omics resource defines circadian signatures of inverted feeding in peripheral tissues (www.CircaMetDB.org.cn).

Zinc finger and BTB domain containing 7A (ZBTB7), a POZ/BTB and Krüppel erythroid myeloid oncogenic factor, is critical for the tumorigenicity and progression of various cancer types. ZBTB7 has been reported to promote the cell proliferation of colorectal cancers (CRC). However, the function of ZBTB7 to 5-fluorouracil (5‑FU) resistance has not yet been studied. In the current study, ZBTB7 expression and function in 5‑FU resistance in CRC were investigated using with multidisciplinary approaches, including western blot analysis, Transwell assay, CCK8 and a tumor xenograft model. Overexpression of ZBTB7 was increased the level of proteins associated with cell invasion and epithelial-mesenchymal transition. ZBTB7 inhibition attenuated the invasion and enhanced the apoptosis of CRC cells. IC50 values and cell viability were significantly reduced in cells with short hairpin RNA (shRNA)-mediated ZBTB7 depletion compared with the control group. 5‑FU administration decreased viability to a greater extent in the ZBTB7-shRNA group compared with the control, which was dose- and time-dependent. Analysis of gene expression omnibus data demonstrated that ZBTB7 mediated 5‑FU resistance, potentially through nuclear factor (NF)-κB signaling. NF‑κB inhibitor SN50 reversed ZBTB7-induced resistance in CRC. Collectively, the findings demonstrated that ZBTB7 mediated 5‑FU resistance in CRC cells through NF‑κB signaling. Thus, targeting ZBTB7 and NF‑κB signaling may be an effective strategy to reverse 5‑FU resistance in CRC.

Surveys were conducted in four seasons in the eastern Beibu Gulf from July 2006 to November 2007, to determine the ichthyoplankton composition, abundance, as well as environmental factors impacted on their spatiotemporal distributions. The fish eggs and larvae were sorted from 303 zooplankton quantitative samples, in which at least 1 order, 60 families, 31 genera and 61 species of fish eggs and larvae were identified. The species number was highest in summer whereas lowest in winter. With the most abundant fish egg (the average density was 2.41 ind./m3), spring was the main spawning season, while the greatest mean density of fish larva was 1.35 ind./m3 in summer. In the whole year around, fish eggs had the trends to aggregate in the nearshore of Guangxi coast, fish larvae were more abundant in the northern water. Relationships between fish eggs and larvae abundance and environmental factors were analyzed by using the Yield-Density model. The spatiotemporal distribution of ichthyoplankton in eastern Beibu Gulf was closely related to the Chl a concentration, and the optimal temperature, salinity and Chl a were 19.4–21.7°C, 31.8–33.1 and 1.5–4.8 mg/m3, respectively. Additionally, the distribution of fish larvae could be effected by ocean currents in summer and autumn.

microRNAs (miRNAs) have been documented playing a critical role in cancer development and progression. In this study, we investigated the role of miR-548l in non-small cell lung cancer (NSCLC) migration and invasion. microRNAs microarray analysis was used to detect the differentially expressed miRNAs between various metastatic levels of NSCLC cells and further confirmed by real-time PCR analysis. To facilitate the delineation of the role of selected miR-548l in NSCLC pathology, we detected its expression in 22 NSCLC tissues. Proliferation, apoptosis, invasion and metastasis effects of the miRNA were evaluated using MTT, flow cytometry, wound healing and invasion assay following transfection with mimics and inhibitors. Luciferase assay and Western blot analysis were performed to assess miR-548l binding to AKT1 gene. AKT1 expression in the clinical tissues was evaluated using immunohistochemical staining. The results showed a negative relationship between miR-548l expression and lymph node metastasis of NSCLC. Functional assays showed that over-expression of miR-548l suppressed NSCLC cell migration and invasion. Luciferase assays confirmed that miR-548l could directly bind to the 3′ untranslated region of AKT1. Further data showed that the over-expression of AKT1 could rescue the effects of miR-548l in NSCLC cells, and the miR-548l expression was inversely correlated with AKT1 expression in NSCLC tissues. These results indicated that AKT1 was involved in miR-548l-induced suppression of NSCLC cell migration and invasion. These results suggested that miR-548l may play a causal role through AKT1 in NSCLC invasion and metastasis.

Abstract 4929 Background Multiple myeloma myeloma accounts for approximately 1 percent of all cancers and 10 percent of hematologic cancers. It is incurable with conventional chemotherapy. Recent studies suggest that rHuEpo has additional biological effects which significantly prolonged the expected survival of six end-stage multiple myeloma (MM) patients and led to hypothesize that Epo might have an anti-neoplastic or anti-myeloma effect. This latter observation was further supported by studies. Objective Based on the myeloma models which established in BALB/c murine with MPC-11 myeloma cell line, our study is to verify the true effects, to research the suitable dose and administration way of recombinant human erythropietin(rHuEPO) for individuals with MM, as well as the possible antitumor mechanism in BALB/c murine myeloma models. Methods 550 healthful BALB/c mice were injected s.c. with 6×105 MPC-11 cells to establish the multiple myeloma models. When small palpable tumors were observed, the tumor-bearing mice were randomizedly divided into five groups which were respectively administrated the various rHuEPO doses (10,20,30,40 and 50 units per injection) and a placebo group with the saline. In the study, rHuEPO treatment was strated in 5 days after the initial MPC-11 cells injection, daily s.c. injections of rHuEPO for 30 consecutive days, followed by one injection per week for 4 weeks with a same dose as before. Hemoglobin, serum IL-6 and TNF-α were monitored before the start of rHuEPO administration and in 15 days, 30 days and 60 days after the start of rHuEPO administration. Immunofixation electrophoresis(IFE) was performed to detect the existence of the serum monoclonal immunoglobulin before the rHuEPO treatment and in 60 days after the rHuEPO-treatment. In the second part of the study, microvessel density(MVD) and TdT-mediated dUTP nick end labeling (TUNEL) techniques were performed to evaluate the tumor proliferation burden and tumor cell apoptosis respectively. Tumor volume change was monitored twice a week and weight change of tumour-bearing mice was measured once a week for every individual mouse during the whole study. Results The serum monoclonal immunoglobulin appeared in 12 days after the initial MPC-11 cells injection. There wasn't a significant decrease in tumor size of bearing cancer mice after administration of rHuEPO, and there was no significant difference among each dosage group. rHuEPO can obviously increase Hb levels in bearing cancer mice. The mice with a smaller tumor size had a higher Hb level, which resulting in a longer survival. Individual survival showed a significant positive correlation with Hb level (Spearman's rho 0.821, p=0.000), and in contrast, it had a negative correlation with tumor size (Spearman's rho 0.475, p=0.009). The serum levels of IL-6 showed no significant differences between the each rHuEPO treatment group and the placebo group, and the result was the same in the serum levels of TNF-α. Also, there is no difference in MVD between the treatment groups and placebo group. The apoptotic index of the treatment groups with rHuEPO seemed a little higher than that of the placebo group. Conclusion rHuEPO administration to the tumor-bearing mice seemed to prolong their individual survival. Insufficient evidence was found for the anti-MM therapeutic role of rHuEPO in our study. Disclosures No relevant conflicts of interest to declare.