The Long Terminal Repeats of ERV6 Are Activated in Pre-Implantation Embryos of Cynomolgus Monkey.

Precise gene regulation is critical during embryo development. Long terminal repeat elements (LTRs) of endogenous retroviruses (ERVs) are dynamically expressed in blastocysts of mammalian embryos. However, the expression pattern of LTRs in monkey blastocyst is still unknown. By single-cell RNA-sequencing (seq) data of cynomolgus monkeys, we found that LTRs of several ERV families, including MacERV6, MacERV3, MacERV2, MacERVK1, and MacERVK2, were highly expressed in pre-implantation embryo cells including epiblast (EPI), trophectoderm (TrB), and primitive endoderm (PrE), but were depleted in post-implantation. We knocked down MacERV6-LTR1a in cynomolgus monkeys with a short hairpin RNA (shRNA) strategy to examine the potential function of MacERV6-LTR1a in the early development of monkey embryos. The silence of MacERV6-LTR1a mainly postpones the differentiation of TrB, EPI, and PrE cells in embryos at day 7 compared to control. Moreover, we confirmed MacERV6-LTR1a could recruit Estrogen Related Receptor Beta (ESRRB), which plays an important role in the maintenance of self-renewal and pluripotency of embryonic and trophoblast stem cells through different signaling pathways including FGF and Wnt signaling pathways. In summary, these results suggest that MacERV6-LTR1a is involved in gene regulation of the pre-implantation embryo of the cynomolgus monkeys.

[Differential proteins in esophageal squamous cell line EC9706/CDDP identified by SILAC quantitative proteomic approach].

Multidrug resistance (MDR) is one of the main causes leading to the failure in cancer treatment. Differential proteins between esophageal squamous cell carcinoma (ESCC) cell line EC9706 and its cisdiamminedichloroplatinum (CDDP)-resistant subline EC9706/CDDP revealed by quantitative analysis may provide deeper insights into the molecular mechanisms of MDR implicated in ESCC. EC9706/CDDP was generated by exposure of its parental sensitive EC9706 to a step-wise increase of CDDP concentration during EC9706 cultivation. The stable isotope labeling with amino acids in cell culture (SILAC) was used to label EC9706 and EC9706/CDDP with heavy and light medium, separately. Mixed peptides derived from EC9706 and EC9706/CDDP were analyzed by high performance liquid chromatography-electrospray ionization-mass spectrometry (HPLC-ESI-MS/MS) and subsequently subjected to bioinformatics analysis to identify differential proteins between EC9706 and EC9706/CDDP. Compared to parental EC9706, EC9706/CDDP manifested phenotypes of slow proliferation, cell pleomorphology, atypia and increased resistant-index 3.23. Seventy-four differential proteins identified in the present study belongs to various families with multiple functions, such as cytoskeleton (20%), energy metabolism (11%), transcription regulation and DNA repair (11%), redox homeostasis (9.5%), protein biosynthesis and mRNA processing (12%), ribosome constituent (8.1%), molecular chaperone (8.1%), immunity/inflammation (5.4%), intracellular transport (5.4%) and nucleosome assembly (2.7%), which indicated that development of MDR is a complicated process involving dysregulation of multiple molecules and pathways. The data is of great value for in-depth elucidation of molecular mechanisms of the MDR implicated in ESCC and may represent potential molecular targets for future therapeutic development.

The establishment and characterization of adriamycin-resistant cell lines derived from Saos-2.

BACKGROUND: The aim was to establish and characterize adriamycin (ADM)-resistant cell lines. MATERIAL/METHODS: Two cell lines, named Saos-2/ADM1 and Saos-2/ADM4, resistant to ADM, were established after 167 days. RESULTS: The resistance indices of methotrexate for the Saos-2/ADM1 and Saos-2/ADM4 cell lines to ADM were 49.8 and 74.6 times higher, respectively, than that of Saos-2. The two cells lines had resistance to MTX, IFO, EPI, THP, and PTX, while the cells were found to remain sensitive to cisplatin. Disordered and coenocytic structure were observed via microscopy. CONCLUSIONS: This is the first report on ADM-resistant cells based on clinical peak doses in serum. MDR1 and MRP genes are involved in the resistance of cell lines to ADM, which are invaluable tools to study the resistance of anticancer drugs and to find the means to revert the resistance.

Bis{tris-[3-(2-pyrid-yl)pyrazole]manganese(II)} dodeca-molybdosilicate hexa-hydrate.

The title compound, [Mn(C(8)H(7)N(3))(3)](2)[SiMo(12)O(40)]·6H(2)O, consists of an [SiMo(12)O(40)](4-) heteropolyanion, lying on a centre of inversion, and a complex [Mn(C(8)H(7)N(3))(3)](4+) cation. The Mn(II) atom of the cation is hexa-coordinated in a distorted octa-hedral geometry by six N atoms from three chelating 3-(2-pyrid-yl)pyrazole ligands. In the heteropolyanion, the four O atoms of the tetra-hedral SiO(4) group each half-occupy eight sites due to Si lying on the centre of inversion. N-H⋯O and O-H⋯O hydrogen bonding mediated by the water mol-ecules leads to a consolidation of the structure.