Search

Aim: To explore the effect of miR-382 on esophageal squamous cell carcinoma (ESCC) in vitro and its possible molecular mechanism., Methods: Eca109 cells derived from human ESCC and Het-1A cells derived from human normal esophageal epithelium were used. Lentivirus-mediated miR-382 was overexpressed in Eca109 cells. The effect of miR-382 on cell proliferation was evaluated by MTT and colony formation assay. For cell cycle analysis, cells were fixed and stained for 30 min with propidium iodide (PI) staining buffer containing 10 mg/mL PI and 100 mg/mL RNase A, and analyzed by BD FACSCalibur™ flow cytometer. For cell apoptosis assay, cells were stained with an Annexin V-FITC/PI Apoptosis Detection Kit according to the manufacturer's instructions and analyzed by a dual-laser flow cytometer. Cell invasion and migration abilities were determined through use of transwell chambers, non-coated or pre-coated with matrigel. Levels of proteins related to cell growth and migration were examined by western blotting., Results: Endogenous miR-382 was down-regulated in Eca109 cells compared with Het-1A. Introduction of miR-382 not only significantly inhibited proliferation and colony formation, but also arrested cell cycle at the G2/M phase, as well as promoted apoptosis and autophagy in Eca109 cells. Migration, invasion and epithelial-mesenchymal transition of Eca109 cells were suppressed by overexpressing miR-382. Western blotting results showed that miR-382 inhibited the phosphorylation of mTOR and 4E-BP1., Conclusion: miR-382 functions as a tumor suppressor against ESCC development and metastasis, and could be considered as a potential drug source for the treatment of ESCC patients., Competing Interests: Conflict-of-interest statement: The authors declared that no conflict of interest exists in this study.

Two motile actinomycetes, designated strains 03-723(T) and 03-8772(T), which had potent inhibitory activity against Enterococcus faecium peptide deformylase and several clinical Gram-positive, antibiotic-resistant strains, were isolated from two soil samples collected from Sichuan Province and Xinjiang Uyghur Autonomous Region in China, respectively. The taxonomic status of these two organisms was established by using a polyphasic approach. The taxonomic data were consistent with the assignment of the strains to the genus Actinoplanes. The neighbour-joining phylogenetic tree based on 16S rRNA gene sequences showed that the two isolates formed a branch with the type strains of Actinoplanes lobatus, Actinoplanes auranticolor, Actinoplanes capillaceus, Actinoplanes campanulatus and Actinoplanes philippinensis in the clade of Actinoplanes species. This branching pattern was also supported by the tree constructed with the maximum-parsimony method. Levels of 16S rRNA gene sequence similarity between strains 03-723(T) and 03-8772(T) and their phylogenetic neighbours ranged from 98.0 to 98.8 % and 97.4 to 98.1 %, respectively. However, the two strains shared low levels of DNA-DNA relatedness with the type strains of closely related Actinoplanes species and were readily distinguished by using a combination of phenotypic properties. Therefore, it is proposed that strains 03-723(T) and 03-8772(T) represent two novel species of the genus Actinoplanes, for which the names Actinoplanes sichuanensis sp. nov. (type strain 03-723(T)=KCTC 19460(T)=CCM 7526(T)) and Actinoplanes xinjiangensis sp. nov. (type strain 03-8772(T)=KCTC 19461(T)=CCM 7527(T)) are proposed.

Three Gram-positive, aerobic, non-motile, non-acid-alcohol-fast strains, designated I05-00051, I05-00074T and I03-00808, were isolated from different soil samples in Beijing and Sichuan, China. Phylogenetic analysis based on 16S rRNA gene sequences and DNA-DNA hybridization experiments revealed that these three isolates represented the same genospecies. These three strains showed <97.0 % 16S rRNA gene sequence similarity with the type strains of recognized species of the genus Saccharopolyspora, with the exception of Saccharopolyspora hirsuta subsp. hirsuta DSM 43463T (98.1 % gene sequence similarity) and Saccharopolyspora spinosa DSM 44228T (98.0 % similarity). Chemotaxonomic data, including meso-diaminopimelic acid as the diagnostic diamino acid, arabinose and galactose as predominant sugars, iso-C15 : 0, iso-C16 : 0, iso-C17 : 0 and anteiso-C17 : 0 as major fatty acids, MK-9(H4) as predominant menaquinone and polar lipids dominated by diphosphatidylglycerol, phosphatidylcholine, phosphatidylglycerol and phosphatidylinositol, supported the affiliation of these three organisms to the genus Saccharopolyspora. The genomic DNA G+C contents of the three isolates were 68.2-69.9 mol%. The results of DNA-DNA hybridization experiments among these three isolates and S. hirsuta subsp. hirsuta DSM 43463T and S. spinosa DSM 44228T, in combination with chemotaxonomic and physiological data, demonstrated that the three new isolates represent a novel species of the genus Saccharopolyspora, for which the name Saccharopolyspora antimicrobica sp. nov. is proposed. The type strain is I05-00074T (=CCM 7463T=KCTC 19303T).