Angiogenesis is considered as an important process in the development of malignancies and is associated with cancer progression and metastasis. Hepatocellular carcinoma (HCC) is the most common primary tumor of the liver and is recognized as a typical angiogenic tumor. Thus, it is of great importance to study the underlying mechanism of angiogenesis in HCC. The long non-coding RNA (lncRNA) ubiquitin conjugating enzyme E2C pseudogene 3 (UBE2CP3) has been reported as an oncogene that promotes tumor metastasis in HCC. However, the role and underlying mechanisms of UBE2CP3 in HCC angiogenesis are still unclear. We measured the expression levels of UBE2CP3 by in situ hybridization (ISH) and quantitative real-time polymerase chain reaction (qRT-PCR) in HCC patient samples. We also concomitantly used CD31/PAS double-staining to measure endothelial vessel (EV) density and used qRT-PCR to measure the CD31 mRNA level. HepG2 and SMMC-7721 cells were transfected with Lv-UBE2CP3 or Sh-UBE2CP3 virus to obtain stably over-expressing or knocking-down UBE2CP3 cell lines. The indirect effects of UBE2CP3 on ECs were studied by establishing a co-culture system using Transwell chambers with a 0.4-μm pore size. HCC cells and ECs in the co-culture system were separated, but the cytokines and growth factors were able to communicate with each other. Following exposed to HCC cells, ECs were collected for functional studies. Finally, we studied the function of UBE2CP3 in vivo by chick embryo chorioallantoic membrane (CAM) angiogenesis assays and nude mouse tumorigenicity assays. In this study, we found that UBE2CP3 expression was higher in HCC tissues than in para-tumor tissues and was up-regulated in tissues with high EV density. Functionally, we found that in the co-culture systems, HCC cells overexpressing UBE2CP3 promoted HUVEC proliferation, migration and tube formation via the activation of ERK/HIF-1α/p70S6K/VEGFA signalling, increasing the level of VEGFA in HCC cell supernatant. In addition, the opposite results appeared when the expression of UBE2CP3 in HCC cells was knocked down. Consistent with these results, CAM angiogenesis assays and nude mouse tumorigenicity assays showed that UBE2CP3 expression up-regulated EV density in vivo. Our study suggests that UBE2CP3 can enhance the interaction between HCC tumor cells and HUVECs and promote HCC tumorigenicity by facilitating angiogenesis.

Insulin resistance and associated metabolic health symptoms remain a primary global health concern. In addition to healthy dietary and nutritional programs, sleep duration is closely related to and has been linked to healthy metabolism. This study aimed to determine the link between insulin resistance and sleep duration and the dietary intake of live microbes. Data were collected from 15,927 participants in the National Health and Nutrition Examination Survey database from 2005 to 2018; this sample is equivalent to 209,316,590 individuals in the United States. The participants were categorized according to their consumption of foods containing live microbes: low, medium, high, and medium-high. The relationship between diets containing live microbes and the triglyceride-glucose index was analysed using a weighted multivariate linear regression model with a multistage sampling approach. The individuals were deemed to have insulin resistance if their homeostatic model assessment score for insulin resistance was ≥ 2. The relationship between diets containing live microbes and insulin resistance status was assessed using weighted multivariate logistic regression analyses. The mediating role of sleep duration on the relationship between diets containing live microbes and the triglyceride-glucose index was also examined. After accounting for potential confounders, diets containing live microbes at medium and medium-high levels were significantly associated with a reduced triglyceride-glucose index. The medium and medium-high levels of live microbial intake were also associated with a lower risk of insulin resistance. Within the 6-9 hours' sleep duration range, the indirect effect of medium and medium-high levels of live microbes on the triglyceride-glucose index was observed, accounting for 2.95% and 6.08% of the overall change, respectively. This study suggests an association between a diet rich in medium and medium-high viable microbes, lower triglyceride-glucose index values, and a reduced risk of developing insulin resistance. Additionally, a sleep duration of 6-9 h may mediate this association.