Global identification of circular RNAs in chronic myeloid leukemia reveals hsa_circ_0080145 regulates cell proliferation by sponging miR-29b.

Circular RNAs (circRNAs) are a class of non-coding RNAs that participate in various biological processes and disease pathogenesis. However, the role of circRNAs in chronic myeloid leukemia (CML) remains largely unknown. In this study, high-throughput sequencing was performed to explore the expression profile of circRNAs in CML patients for the first time, and a large number of differentially expressed circRNAs were identified. In addition, we constructed potential circRNA-miRNA interaction networks in CML, including a detailed hsa_circ_0080145-mediated competing endogenous RNA (ceRNA) regulatory network. Furthermore, functional experiments demonstrated that hsa_circ_0080145 knockdown significantly suppressed CML cell proliferation and hsa_circ_0080145 regulated CML cell proliferation by acting as an miR-29b sponge.

Association between the expression levels of ADAMTS16 and BMP2 and tumor budding in hepatocellular carcinoma.

Tumor budding (TB) has become a crucial factor for predicting the malignancy grade and prognostic outcome for multiple types of solid cancer. Studies have investigated the prognostic value of TB in hepatocellular carcinoma (HCC). However, its molecular mechanism in HCC remains unclear. To the best of our knowledge, the present study was the first to compare the expression of differentially expressed genes (DEGs) between TB-positive (TB-pos) and TB-negative HCC tissues. In the present study, total RNA was extracted from 40 HCC tissue specimens and then sequenced. According to Gene Ontology (GO) functional annotation, upregulated DEGs were markedly associated with embryonic kidney development-related GO terms, which suggested that the TB process may at least partly mimic the process of embryonic kidney development. Subsequently, two genes, a disintegrin and metalloproteinase domain with thrombospondin motifs 16 (ADAMTS16) and bone morphogenetic protein 2 (BMP2), were screened and verified through immunohistochemical analysis of HCC tissue microarrays. According to the immunohistochemical results, ADAMTS16 and BMP2 were upregulated in TB-pos HCC samples, and BMP2 expression was increased in budding cells compared with the tumor center. Additionally, through cell culture experiments, it was demonstrated that ADAMTS16 and BMP2 may promote TB of liver cancer, thus promoting the malignant progression of liver cancer. Further analysis revealed that ADAMTS16 expression was associated with necrosis and cholestasis, and BMP2 expression was associated with the Barcelona Clinic Liver Cancer stage and the vessels encapsulating tumor clusters. Overall, the findings of the present study provided insights into the possible mechanisms of TB in HCC and revealed potential anti-HCC therapeutic targets.

Case report: ZEB1 expression in three cases of hepatic carcinosarcoma.

Hepatic carcinosarcoma (HCS) is defined as a tumor that contains cancer from the epithelium and sarcoma from mesenchymal tissue. HCS has a low incidence rate and is composed of osteosarcoma, chondrosarcoma, or angiosarcoma. Though surgery is the main treatment for HCS, it has proven unsatisfactory, resulting in a very poor prognosis of HCS. Currently, the reports on HCS are mainly about the description of clinical pathological phenomena, imaging features, and mutation sites of related genes, the underlying molecular mechanism of HCS remains undefined. Through the dynamic process of epithelial-mesenchymal transition (EMT), cancer cells acquire a mesenchymal phenotype, simultaneously losing epithelial properties. Zinc finger E-box binding homeobox 1 (ZEB1) is an EMT-inducing transcription factor; its main regulatory target is E-cadherin in EMT process. Esophageal carcinosarcoma (ECS) is associated with EMT. The current study showed that EMT might promote the development of ECS and uterine carcinosarcoma (UCS), and ZEB1 was highly expressed in the sarcomatous components. In the current study, three cases were collected, and the clinicopathological features were compared with those of corresponding cases. The expression level, and subcellular localization of ZEB1 were detected using immunohistochemistry. The expression of the ZEB1 in the nucleus was found to be significantly higher in sarcomatous components than that in cancer components in all three cases, suggesting an association of HCS with EMT.

Evaluation of Epstein-Barr virus latent membrane protein 2 specific T-cell receptors driven by T-cell specific promoters using lentiviral vector.

Transduction of latent membrane protein 2 (LMP2)-specific T-cell receptors into activated T lymphocytes may provide a universal, MHC-restricted mean to treat EBV-associated tumors in adoptive immunotherapy. We compared TCR-specific promoters of distinct origin in lentiviral vectors, that is, Vß6.7, delta, luria, and Vß5.1 to evaluate TCR gene expression in human primary peripheral blood monocytes and T cell line HSB2. Vectors containing Vß 6.7 promoter were found to be optimal for expression in PBMCs, and they maintained expression of the transduced TCRs for up to 7 weeks. These cells had the potential to recognize subdominant EBV latency antigens as measured by cytotoxicity and IFN-γ secretion. The nude mice also exhibited significant resistance to the HLA-A2 and LMP2-positive CNE tumor cell challenge after being infused with lentiviral transduced CTLs. In conclusion, LMP2-specific CTLs by lentiviral transduction have the potential use for treatment of EBV-related tumors.

Hydrodynamics-based transfection of plasmid encoding receptor activator for nuclear factor kappa B-Fc protects against hepatic ischemia/reperfusion injury in mice.

Hepatic ischemia/reperfusion (I/R) injury is very important in transplant surgery. To study the mechanism of receptor activator for nuclear factor kappa B-Fc (RANK-Fc) in protection against I/R injury, 90 male BALB/c mice were randomly divided into 3 groups: a phosphate-buffered saline (PBS) (sham) group, a pLNCX2-IRES-eGFP+I/R (Negative-control) group (where IRES means internal ribosome entry site and eGFP means enhanced green fluorescent protein), and a pLNCX2-RANK-Fc-IRES-eGFP+I/R (RANK-Fc) group. All mice were injected with 2.5 mL of PBS (with or without plasmids) within 6 seconds via the tail vein. After 3 days, hepatic I/R was induced under warm conditions by partial occlusion of the left and median lobes for 90 minutes followed by various periods of reperfusion. Hepatic injury was assessed by the levels of liver aminotransferases and histopathology. Tumor necrosis factor alpha, interleukin 6, and interleukin 1beta were measured by enzyme-linked immunosorbent assay, whereas RANK-Fc, phospho-c-Jun, c-Jun N-terminal kinase (JNK), hypoxia inducible factor 1 alpha (HIF-1alpha), nuclear p65, and total p65 were assessed with western blotting. Apoptosis was identified by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling. RANK-Fc was efficiently expressed in the liver. In comparison with the negative-control group, RANK-Fc reduced nuclear factor kappa B (NF-kappaB) p65 nuclear translocation, JNK phosphorylation, and HIF-1alpha expression during I/R. RANK-Fc effectively suppressed proinflammatory cytokine expression. The results indicated that RANK-Fc could protect against hepatic I/R injury in mice at least in part via the inhibition of the proinflammatory NF-kappaB pathway as well as proapoptotic JNK and HIF-1alpha pathway activation.

Key role of P38 mitogen-activated protein kinase and the lipoxygenase pathway in angiotensin II actions in H295R adrenocortical cells.

Angiotensin II (ANG II) can activate the mitogen-activated protein kinases (MAPKs) and stress-activated protein kinases in several cell types. We have previously shown that the 12-lipoxygenase (12-LO) pathway of arachidonic acid metabolism is a mediator of ANG II-induced aldosterone synthesis in adrenal glomerulosa cells. To evaluate the role of MAPK activation in ANG II and the effects of LO on aldosterone synthesis, experiments were performed using the human adrenocortical cell line H295R, which secretes aldosterone in response to ANG II. MAPK activities were determined by Western immunoblotting using specific antibodies to their activated phosphorylated forms. ANG II led to a dose-dependent increase in extracellular signal-regulated kinase (ERK1/2) activity in these cells, with a peak at 5 min and lasting up to 3 h. The effects of ANG II were blocked by the ANG-II Type 1 receptor antagonist losartan. A specific 12-LO product, 12(S)-hydroxyeicosatetraenoic acid (12-HETE), had no direct effect on ERK activity. However, both ANG II and 12-HETE led to significant dose-dependent increases in p38 MAPK activity with peak effects at 5 min. By contrast, the 15-LO product, 15-HETE, had no effect on p38 MAPK activity. Furthermore, two dissimilar 12-LO inhibitors, CDC and baicalein, blocked ANG II-induced p38 MAPK activation. ANG II significantly increased aldosterone release, and this effect was inhibited by the LO inhibitor baicalein, as well as a specific p38 MAPK inhibitor, SB202190, but not by PD098059, a specific inhibitor of the ERK activator MEK. In summary, in H295R cells, ANG II activated ERK and p38 MAPKs, ANG II-induced p38 MAPK was mediated by 12-LO activation, and ANG II-induced aldosterone synthesis was prevented by 12-LO- and p38 MAPK-specific inhibitors. These results suggest, for the first time, that activation of p38 MAPK, either directly or via LO activation, participates in aldosterone's stimulatory effects of ANG II in adrenal cells.