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This study aimed to investigate the role of miRNA-1225-5p (miR-1225) in laryngeal carcinoma (LC). We found that the expression of miR-1225 was suppressed in human LC samples, while CDC14B (cell division cycle 14B) expression was reinforced in comparison with surrounding normal tissues. We also demonstrated that enhanced expression of miR-1225 impaired the proliferation and survival of LC cells, and resulted in G1/S cell cycle arrest. In contrast, reduced expression of miR-1225 promoted cell survival. Moreover, miR-1225 resulted in G1/S cell cycle arrest and enhanced cell death. Further, miR-1225 targets CDC14B 3′-UTR and recovery of CDC14B expression counteracted the suppressive influence of miR-1225 on LC cells. Thus, these findings offer insight into the biological and molecular mechanisms behind the development of LC. [ABSTRACT FROM AUTHOR]

Background: At present, few studies have explored the significance of POU5F1 (also known as octamer-bingding factor, Oct-4 or Oct-3) expression in breast cancer tissues.Methods: A total of 121 patients were retrospectively selected between May 2010 and March 2013 to investigate the relationship between POU5F1/Oct-4 expression in breast cancer tissues and non-sentinel lymph node (non-SLN) metastases and to validate the Memorial Sloan-Kettering Cancer Center (MSKCC) nomogram. All patients had early-stage breast cancer, which was histologically confirmed by the Department of Surgical Oncology, The First Affiliated Hospital of China Medical University. Histological type and grade of tumors were determined from tissue samples by hematoxylin and eosin staining, while the presence of POU5F1/Oct-4 protein was determined by immunohistochemistry. POU5F1/Oct-4 expression levels in tissues obtained from patients with sentinel lymph node (SLN) and non-SLN metastasis and in tissues obtained from patients without lymph node metastases were compared.Results: POU5F1/Oct-4 expression levels in breast cancer tissues were significantly higher in both the SLN metastasis and non-SLN metastasis groups (P = 0.003 and P = 0.030, respectively). Furthermore, POU5F1/Oct-4 expression was found to be associated to both histological (P = 0.01) and molecular type (P = 0.03). Thus, our data once again confirms the validity of the MSKCC nomogram. The area under curve (AUC) was 0.919 (95 % CI: 0.869-0.969, P < 0.001). The probability of non-SLN metastasis generated from the MSKCC nomgram was significantly higher in the POU5F1/Oct-4 positive group than in the POU5F1/Oct-4 negative group. Both univariate and multivariate analysis revealed that Oct-4 expression levels were significantly associated with non-SLN metastases (P = 0.030 and P = 0.034, respectively).Conclusions: POU5F1/Oct-4 expression levels are significantly associated with non-SLN metastases. Patients with higher probabilities of metastasis generated from the MSKCC nomogram may also have higher POU5F1/Oct-4 expression levels. [ABSTRACT FROM AUTHOR]

Highlights: [•] Generation of Bm5 transformed cell line that expresses BmToll9-1. [•] Higher transcriptional response of Dcr2 by dsRNA in BmToll9-1 expressing cells. [•] Lower transcriptional response of innate immune pathway and AMP genes to LPS in BmToll9-1 expressing cells. [•] No direct modulation of RNAi process by BmToll9-1. [ABSTRACT FROM AUTHOR]

Highlights: [•] Dominant expression of BmToll9-1 mRNA in the larval and pupal midgut tissues. [•] High breakdown of dsRNA integrity in the insect midgut. [•] Injection of dsRNA suppresses expression of BmToll9-1 mRNA in the larval midgut. [•] Induction of BmDcr2 and BmAgo2 transcription in the larval midgut by dsRNA. [Copyright &y& Elsevier]

Abstract: A DNA/RNA non-specific alkaline nuclease (BmdsRNase) was isolated from the digestive juice of Bombyx mori. While originally reported to be produced by the midgut only, in this project it was found that the mRNA of this enzyme was also expressed in the epidermis, fat body, gut, thoracic muscles, Malpighian tubules, brain, and silk glands of 5th instar larvae, indicating additional functions to its reported role in nucleic acid digestion in the midgut. In order to study the functional properties of BmdsRNase, three pEA-BmdsRNase expression constructs were generated, characterized by presence or absence of a signal peptide and a propeptide, and used for expression in lepidopteran Hi5 tissue culture cells. Western blot indicated that these different forms of BmdsRNase protein were not secreted into the growth medium, while they were detected in the pellets and supernatants of Hi5 cell extracts. Nucleic acids cleavage experiments indicated that full-length BmdsRNase could digest dsRNA and that the processed form (absence of signal peptide and propeptide) of BmdsRNase could degrade both DNA and dsRNA in Hi5 cell culture. Using a reporter assay targeted by transfected homologous dsRNA, it was shown that the digestive property of the processed form could interfere with the RNAi response. Immunostaining of processed BmdsRNase protein showed asymmetric localization in the cellular cytoplasm and co-localization with Flag-tagged Dicer-2 was also observed. In conclusion, our in vitro studies indicated that intracellular protein isoforms of BmdsRNase can be functional and involved in the regulation of nucleic acid metabolism in the cytoplasm. In particular, because of its propensity to degrade dsRNA, the enzyme might be involved in the innate immune response against invading nucleic acids such as RNA viruses. [Copyright &y& Elsevier]