TIMP-1 suppressed by miR-138 participates in endoplasmic reticulum stress-induced osteoblast apoptosis in osteoporosis

The aim of this study was to investigate the role of miR-138 in osteoporosis and its underlying mechanism. Hydrogen peroxide (H₂O₂) was used to induce osteoporotic injury of osteoblasts. The cell viability and apoptosis of MC3T3-E1 cells was assessed using MTT assay and flow cytometry, respectively. The cell transfection was carried out to modulate the expression levels of miR-138 and TIMP-1 in MC3T3-E1 cells. Luciferase reporter gene assay was performed to determine the interaction between miR-138 and TIMP-1 3′UTR. In the present study, H₂O₂ inhibited osteoblasts growth and induced intracellular endoplasmic reticulum (ER) stress accompanied by high expression of miR-138. We also confirmed that miR-138 promoted osteoblasts apoptosis in vitro and in vivo. MiR-138 was further indicated to inhibit osteoblast survival via negative regulating TIMP-1 expression. Moreover, the downregulated TIMP-1 also mediated the ER stress-induced apoptosis of osteoblasts. We confirmed that miR-138 and ER stress were induced in osteoporosis and then promoted the apoptosis of osteoblasts, at least in part, through TIMP-1.     

MiR-96 enhances cellular proliferation and tumorigenicity of human cervical carcinoma cells through PTPN9

Up to date, the cervical cancer remains to be one of the leading gynecological malignancies worldwide. MicroRNAs (miRNAs) play critical roles in the process of tumor initiation and progression. However, miR-96 has rarely been investigated in human cervical carcinoma. We aimed to investigate the biological function and underlying molecular mechanism of miR-96 in human cervical carcinoma. MiR-96 levels were determined by qRT-PCR. Protein tyrosine phosphatase, non-receptor type 9 (PTPN9) mRNA and protein levels were investigated by qRT-PCR and western blotting. The cellular proliferation in cervical cells was monitored by CyQuant assay. Soft agar assay was employed to determine the tumorigenicity. 3′ UTR luciferase assay was used to validate the target gene of miR-96. SPSS was used to analyze statistical significance in different treatment. MiR-96 was dramatically upregulated in human cervical tumor tissues. Overexpression of miR-96 was found to significantly promote the cellular proliferation and tumorigenicity of cervical cells. Furthermore, we showed that PTPN9 was a direct target gene of miR-96 and had opposite effect to those of miR-96 on cervical cells. MiR-96 may promote the cellular proliferation and tumorigenicity of cervical cells by silencing PTPN9. Our study highlights an importantly regulatory role of miR-96 and suggests that an appropriate manipulation of miR-96 may be a new treatment of human cervical carcinoma in the future.     

NORAD affects the progression of diabetic nephropathy through targeting miR-520h to upregulate TLR4

To uncover the role of NORAD in the progression of diabetic nephropathy (DN) and the underlying mechanism. Relative levels of NORAD and TLR4 in db/m mice and db/db mice were tested. Meanwhile, their levels in glomerular mesangial cells undergoing high-level (H-MC group) or low-level (L-MC) glucose treatment were determined. Regulatory effects of NORAD and TLR4 on proliferative ability and apoptosis in SV40-MES-13 cells were assessed. The interaction in the regulatory loop NORAD/miR-520h/TLR4 was verified through dual-luciferase reporter gene assay, determination of subcellular distribution and RIP (RNA Immunoprecipitation) assay. At last, potential role of the regulatory loop NORAD/miR-520h/TLR4 in regulating DN was clarified. NORAD and TLR4 were upregulated in db/db mice and SV40-MES-13 cells in H-MC group. Overexpression of them promoted proliferative ability and inhibited apoptosis in SV40-MES-13 cells. MiR-520h was confirmed to bind NORAD and TLR4. NORAD, miR-520h and TLR4 were mainly distributed in cytoplasm, which were enriched in anti-Ago2. The regulatory loop NORAD/miR-520h/TLR4 has been demonstrated to promote the progression of DN. The regulatory loop NORAD/miR-520h/TLR4 promotes the proliferative ability and inhibits apoptosis in glomerular mesangial cells, thus aggravating the progression of DN.     

MiR-18b suppresses high-glucose-induced proliferation in HRECs by targeting IGF-1/IGF1R signaling pathways

MicroRNAs (miRNAs) are important for the proliferation of endothelial cells and have been shown to be involved in diabetic retinopathy (DR). In previous study, we found that miRNAs might play a critical role in hyperglycemia-induced endothelial cell proliferation based on miRNA expression profiling. Here, the roles of microRNA-18b (miR-18b) in the proliferation of human retinal endothelial cells (HRECs) were investigated in an in vitro model of HRECs grown in high glucose. We identified that levels of miR-18b were decreased in high-glucose-induced HRECs, compared with those in cells incubated in normal glucose. However, the reduction of miR-18b up-regulated vascular endothelial growth factor (VEGF) secretion and promoted effects on in vitro proliferation of HRECs. Mechanistically, insulin growth factor-1 (IGF-1) was identified as a target of miR-18b. IGF-1 simulation could antagonize the effect induced by miR-18b up-regulation, promoting cell proliferation and increasing VEGF production. In contrast, the opposite results were observed with silencing IGF-1, which was consistent with the effects of miR-18b overexpression. MiR-18b exerted its function on VEGF synthesis and cell proliferation by suppressing the IGF-1/insulin growth factor-1 receptor (IGF1R) pathway, consequently inhibiting the downstream phosphorylation of Akt, MEK, and ERK. Hence, this may provide a new insight into understanding the mechanism of DR pathogenesis, as well as a potential therapeutic target for proliferative DR.     

Silencing lncRNA DUXAP8 inhibits lung adenocarcinoma progression by targeting miR-26b-5p

Lung adenocarcinoma (LUAD), a common type of lung cancer, has become a popularly aggressive cancer. Long noncoding RNAs (lncRNAs) play a critical role in the pathogenesis of human cancers, while the function of double homeobox A pseudogene 8 (DUXAP8) in LUAD remains to be fully inquired. Therefore, our study was conducted to elucidate the DUXAP8 expression in LUAD and its mechanism on the biological features of LUAD cells. Loss-of-function experiments were performed to assess the function of DUXAP8 proliferation and apoptosis of H1975 and A549 cells. Functionally, silencing DUXAP8 inhibited proliferation and induced apoptosis of LUAD cells. Mechanistically, further correlation assay indicated a negative association between miR-26b-5p and DUXAP8 expression. Subsequently, we testified that DUXAP8 exerted its role in the progression and development of LUAD through targeting miR-26b-5p. In summary, our results elucidated that that DUXAP8 promoted tumor progression in LUAD by targeting miR-26b-5p, which provide a novel therapeutic target for diagnosis and therapy of LUAD.     

Evidence of intraspecific agonistic interactions in Smilodon populator (Carnivora,Felidae)

The saber-toothed cat Smilodon is a characteristic genus of the Pleistocene faunas of the American continent. Smilodon belongs to an extinct clade of felids that had hypertrophied blade-like upper canines. Because the length of the canines is so extreme, the killing bite of Smilodon is a hotly debated topic in vertebrate paleontology. Some authors have proposed that saber-toothed cats had a weak bite and their canines were fragile, not useful for attacking prey or penetrating bones. The aim of the present contribution is to describe two new specimens of Smilodon populator that have injuries on their skulls. Although it cannot be ruled out that the injuries were caused by a potential prey kicking the skull, the size, shape and general features of the injuries suggest that they were inflicted by the upper canines of another Smilodon individual during agonistic interactions.     

Middle to Late Pleistocene herpetofauna from Scladina and Sous-Saint-Paul caves (Namur,Belgium)

For the first time, the fossil herpetofauna from the Middle and Late Pleistocene of Scladina and Sous-Saint-Paul caves (Sclayn, Belgium) is described. The amphibians and squamate reptiles are represented by one salamander (Salamandra salamandra), three anurans (Pelodytes punctatus, Bufo bufo and Rana temporaria), two lizards (Lacerta cf. agilis and Anguis fragilis) and two snakes (Zamenis longissimus and Vipera cf. berus). The occurrence of the Parsley Frog (Pelodytes punctatus) and the Aesculapian Snake (Zamenis longissimus), which are not currently represented in Belgium, is of particular interest. Scladina also represents one of the northernmost fossil mentions for the Fire Salamander (Salamandra salamandra) although it is within its current distribution in Europe. Finally, the presence of the Adder (Vipera cf. berus) is very probably attested in Scladina whereas today this snake is infrequent and classified as endangered in Belgium.     

Fingerprinting and sequence homology of plasmids from different virulent strains of Agrobacterium rhizogenes

Several wild-type virulent strains of Agrobacterium rhizogenes, belonging to both biotypes 1 and 2, were analyzed for plasmid content. All the strains were found to harbor at least one large plasmid, of a size comparable to that of A. tumefaciens Ti plasmids as determined by electron microscopy. The plasmids were characterized by restriction endonuclease finger-printing and compared for sequence homology by the Southern blot-hybridization technique. Despite the diversity of the restriction cleavage patterns, the plasmids of the various strains share rather extensive sequence homology. All of the biotype 1 organisms analyzed were shown to harbor one common plasmid.