Overexpression of miR­214 promotes the progression of human osteosarcoma by regulating the Wnt/ß­catenin signaling pathway.

The aberrant expression of microRNA (miR)­214 contributes to the regulation of normal and cancer cell biology, and is associated with human malignancies, however, it can operate in a contradictory manner. The role of miR­214 in osteosarcoma remains to be fully elucidated. The aim of the present study was to investigate the effects of miR­214 on osteosarcoma progression and tumor cell proliferation, and examine the molecular mechanism underlying osteosarcoma. The level of miR­214 was determined using reverse transcription­quantitative polymerase chain reaction (RT­qPCR) analysis in osteosarcoma and matched paracancerous tissues, and in human osteosarcoma cancer cell lines. The roles of miR­214 in cell proliferation, survival and cell cycle were analyzed using miR­214 lentivirus (LV­miR­214)­infected osteosarcoma cells. In addition, the downstream target proteins in the Wnt/ß­catenin signaling pathway were evaluated using western blot analysis in the LV­miR­214­infected cells. The LV­miR­214­infected MG63 cells were also treated with exogenous ß­catenin for 24, 48 and 72 h, respectively, following which the expression of ß­catenin was measured using western blot analysis and survival was determined using a 3­(4,5­cimethylthiazol­2­yl)­2,5­diphenyl tetrazolium bromide (MTT) assay. The results of the RT­qPCR analysis showed that the expression level of miR­214 was significantly higher in the osteosarcoma tissues, compared with that in the matched paracancerous tissues, and the same was observed in the osteosarcoma cell lines. The MG63, Saos­2 and U2OS cells were infected with the hsa­mir­214 lentivirus for 48 h, and the levels of miR­214 were significantly upregulated in the human osteosarcoma cancer cells. The overexpression of miR­214 in the MG­63 and Saos­2 cells promoted cell growth, and treatment of the cells with specific antisense­microRNA oligonucleotides (AMOs) for miR­214 for indicated durations reversed the effects of miR­214. Additionally, the AMO­treated MG63 cells showed G0/G1 phase arrest, suggesting that miR­214 contributed to regulation of the cell cycle. In addition, the results of western blot analysis showed that, in the miR­214 lentivirus­infected cells, the levels of cyclin­D1, c­myc and lymphoid enhancer­binding factor­1 were significantly increased, compared with those in the control lentivirus­infected cancer cells. Of note, infection with the miR­214 lentivirus did not affect the levels of Wnt1, Wnt2, Wnt4, Axin or glycogen synthase kinase ß in the U2OS cells, whereas the expression levels of ß­catenin in the MG63 cells and Saos­2 cells were significantly increased. The addition of exogenous ß­catenin effectively reversed the efficiency of miR­214­specific AMOs, which was detected using an MTT assay. These data suggested the critical role of miR­214 in human osteosarcoma via regulation of the Wnt/ß­catenin signaling pathway and demonstrated that miR­214 is as an oncogene for human osteosarcoma.

[Effect of Trichinella spiralis and its worm-derived proteins on CLP-induced sepsis in mice].

OBJECTIVE: To observe the effect of Trichinella spiralis and its worm-derived proteins on cecal ligation and puncture (CLP)-induced sepsis in mice. METHODS: Eighty male BALB/c mice were randomly divided into sham-operated group, CLP group, Trichinella spiralis muscle larvae (ML) pre-infection group (ML+CLP group), soluble muscle larvae proteins (SMP) treatment group (SMP+CLP group) and excretory-secretory proteins (MES) treatment group (MES+CLP group). In ML+CLP group, the mice were orally infected with 300 Trichinella spiralis muscle larvae at 28 days before CLP and those in the other groups were intraperitioneally injected with PBS or SMP (25 µg/mice) or MES (25µg/mice) 30 min after CLP. The general condition and 72-h survival after CLP of the mice were observed. The levels of alanine transaminase (ALT), aspartate transaminase (AST), blood urea nitrogen (BUN), creatinine (Cr), TNF-α, IL-6, IL-1ß, IL-10 and TGF-ß were measured at 12 h after the operation, and the pathological changes of the liver and kidney were observed. RESULTS: s Compared with the sham-operated mice, the mice in CLP group showed decreased 72-h survival, obviously increased ALT, AST, BUN, Cr, TNF-α, IL-6, IL-1ß, IL-10, and TGF-ß with hepatic cords disorder, hepatocytes swelling, glomerulus shrinkage, and renal tubular cell edema. Compared with CLP group, the mice in ML+CLP group showed lowered levels of ALT, AST, Cr, TNF-α and IL-1ß and increased levels of IL-10 and TGF-ß; in SMP+CLP group, the levels of ALT, AST, Cr, TNF-α and IL-1ß were decreased and TGF-ß increased. In MES+CLP group, the mice showed obviously increased 72-h survival with lowered levels of ALT, AST, BUN, Cr, TNF-α, IL-6 and IL-1ß, increased levels of IL-10 and TGF-ß, and alleviated liver and kidney damages. CONCLUSION: Trichinella spiralis and its worm-derived proteins can decrease the levels of pro-inflammatory cytokines and increase immunomodulatory cytokines, and MES has more potent effect to reduce structural and functional damages of the liver and kidney.