Search bioRxiv⌕ Search

Biology subjects

XUE, W.

Publications and source records attributed to XUE, W..

3 recordsLinked to original sources

Kinetic measurements of the fluorescent protein synthesis facilitate determination of the miRNA activity.

Oncogenesis is inevitably associated with microRNA expression deregulation. Thus, development of both miRNA targeting substances or small RNA for ant-cancer therapy have been reported. Specifically, repression of the miR-16-1-3p and miR-16-2-3p activities play pivotal roles in osteosarcoma and many other cancers. The majority of miRNA sensors use protein degradation to measure miRNA activities. Here we report miRNA sensors that use fluorescent protein synthesis rather than degradation to measure miRNA activity. Specifically, miR-16-1-3p and miR-16-2-3p sensors consist of the bidirectional tet-On system driving the expression of the Katusha2S protein that is regulated by the RNA interference and GFP as a reference. These sensors specifically detect mir16-1-3p and mir16-2-3p small RNA mimics in the osteosarcoma cell line after doxycycline induction. Kinetic measurements of the reporter responses to the miRNA mimics revealed that pre-induced sensors reach significant differences from the control faster, within 2h than the sensors that were induced after mimic transfection. Thus, kinetic measurements of the fluorescent protein synthesis during doxycycline induction of the tet-On system are feasible for determination of the small RNA activity.

molecular biology↗

The Small Non-coding RNA miR-16-1-3p Hampers Cancer Stem Cell Self-renewal and Invasiveness, Boosting Chemosensitivity by Adjusting TGF-β1 Signaling via MDM2/p53 Axis in Human Osteosarcoma.

The TGF-{beta} signaling pathway has both tumor-suppressing and metastasis-promoting effects in cancer. However, the molecular determinants governing this switch remain unclear. Here, we explored the miR-16-1-3p/MDM2/p53 axis as a critical conductor of the TGF-{beta}-Smad pathway in osteosarcoma. Although miR-16-1-3p overexpression by itself markedly reduces proliferative and clonogenic potential of U2OS cells, when paired with TGF-{beta} treatment, it significantly increases arrest cells in G1 phase and nearly extinguishing the growth capability of these cells. MiR-16-1-3p overexpression inhibited TGF-induced actin remodeling and EMT featuring, significantly decreasing vimentin levels. TGF-{beta} enhances both 2D and 3D migration, but miR-16-1-3p overexpression, alone or with TGF-{beta}, strongly counteracts its pro-migratory effects. MiR-16-1-3p restored p53 stability by targeting MDM2, redirecting TGF-{beta}-Smad signaling toward p21 activation and proliferation inhibition while attenuating its EMT-promoting capacity. Administration of TGF-{beta} together with miR-16-1-3p dramatically increases the sensitivity of wild-type U2OS cells to cisplatin, exceeding that of TGF-{beta} therapy alone by more than an order of magnitude. Administering TGF-{beta} and miR-16-1-3p together significantly reduces the tumor nodule volume and Ki67 expression, while effectively eradicates metastases in the chicken chorioallantoic membrane (CAM) in vivo model. For the first time, our research demonstrates that miR-16-1-3p shifts TGF-{beta}1 signaling from a facilitator of metastasis to a promoter of anti-growth effects through MDM2 inhibition and p53 stabilization, effectively reducing the self-renewal and invasiveness of cancer stem cells in human osteosarcoma model. This process preserves TGF-{beta}s tumor-suppressive role while limiting its associated cancer risks.

cancer biology↗

Butyribacter intestini gen. nov., sp. nov., a butyric acid-producing bacterium of the family Lachnospiraceae isolated from the human faeces, and reclassification of Acetivibrio ethanolgignens as Acetanaerobacter ethanolgignens gen. nov., comb. nov.

A novel, non-motile, Gram-stain-positive, non-spore-forming, obligate anaerobic bacterium, designated strain TF01-11T, was isolated from human faeces. The isolate was characterized by phylogenetic and phenotypic properties, as well as by determination of its whole genome sequence. The growth temperature and pH ranges were 30-42 {degrees}C and 6.0-8.5, respectively. The end products of glucose fermentation were butyric acid and a small amount of acetic acid. The genome was estimated to be 3.61Mbp with G+C content of 36.79 mol%. Genes related to biosynthesis of diaminopimelic acid, polar lipids, polyamines, teichoic and lipoteichoic acids were present. The predominant fatty acids were C16:0 (37.9 %), C14:0 (16.4 %), C13:0 OH/iso-C15:1 H (11.1 %) and C18:1{omega} 9c (10.6 %). Phylogenetic analyses based on 16S rRNA gene sequences, the isolate was a member of family Lachnospiraceae, with the highest sequence similarity to the type strain of Roseburia intestinalis DSM 14610T at 92.18 % followed by Acetivibrio ethanolgignens ATCC 33324T at 91.99 %. The average nucleotide identity (ANI) calculated for the genomes between strain TF01-11T and these closest relatives were 70.5 % and 68.1 %. Based on results of phenotypic characteristics and genotypic properties presented in this study, strain TF01-11T represent a novel species in a new genus, for which the name Butyribacter intestini gen. nov., sp. nov. is proposed. The type strain of the type species is TF01-11T (CGMCC 1.5203T = CGMCC 10984T = DSM 105140T). In addition, Acetivibrio ethanolgignens is proposed to be reclassified as Acetanaerobacter ethanolgignens gen. nov., comb. nov.

microbiology↗