ABSTRACT
Objective To investigate the effect of microRNA (miR) - 140-3p targeting cell division cycle associated 8(CDCA8) on invasion and metastasis of lung adenocarcinoma cells. Methods The differentially expressed miRNAs were analyzed by GE02R in GEO database. The target genes of miR-140-3p were searched by TargetScan human7. 2 and miRWalk databases. The hub gene was screened by Cytoscape 3. 7. 2 software. GEPIA database was used to query the expression levels of target gene in lung adenocarcinoma tissues and normal lung tissues, the expression levels in different stages of lung adenocarcinoma, and the relationship between the expression levels of target gene and the overall survival rate of lung adenocarcinoma patients. The survival analysis of miR-140-3p in lung adenocarcinoma and the correlation between miR-140-3p and CDCA8 expression levels were searched in starBase database. Real-time PCR was used to detect the expression levels of miR-140-3p in normal lung epithelial cells BEAS-2B and lung adenocarcinoma cells A549, as well as the efficiency of infection. Expression levels of CDCA8 mRNA and protein were detected by Real-time PCR and Western blotting experiments after overexpression of miR-140-3p. Dual-luciferase reporter assay verified whether miR-140- 3p directly binds to CDCA8. Transwell invasion assay detected the effect of overexpression of miR-140-3p and CDCA8 on the invasiveness of lung adenocarcinoma cells. Results Analysis result from GEO and other databases showed that the expression level of miR-140-3p in normal lung tissues was significantly higher than that in lung adenocarcinoma, and its predicted target gene CDCA8 expression level in lung adenocarcinoma was significantly higher than that in normal lung tissues, and CDCA8 was negatively correlated with the expression level of miR-140-3p in lung adenocarcinoma. The experimental result showed that the expression of miR-140-3p in A549 cells was significantly lower than that in BEAS-2B cells (P<0.05). The expression level of miR-140-3p increased significantly after lentiviral infection (P<0.05). CDCA8 mRNA and protein expression levels were significantly down-regulated after overexpression of miR-140-3p (P<0.05). Dual-luciferase reporter assay result showed that miR-140-3p could directly bind to CDCA8 (P<0.05). Compared with the control group, overexpression of miR- 140-3p inhibited the invasion and metastasis of lung adenocarcinoma A549, while CDCA8 reversed the inhibition of miR-140-3p on the invasion and metastasis of lung adenocarcinoma A549 (P<0.05). Conclusion MiR-140-3p targeting CDCA8 inhibits the invasion and metastasis of lung adenocarcinoma cells.
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The osteogenic differentiation of mesenchymal stem cells (MSCs) has potential clinical values in the treatmentof bone-related diseases. Long non-coding RNA H19 and microRNA-140-5p (miR-140-5p) have attractedmuch attention of researchers by virtue of their biological importance in cell differentiation and bone formation. Moreover, bioinformatics analyses suggest that miR-140-5p have the potential to bind with H19 andSATB homeobox 2 (SATB2). In this study, we further explored whether H19 could regulate osteogenicdifferentiation of human bone marrow-derived MSCs (BM-MSCs) by miR-140-5p/SATB2 axis. RT-qPCRassay was conducted to examine the expression of H19, miR-140-5p and SATB2. The osteogenic differentiation capacity of BM-MSCs was assessed through alkaline phosphatase (ALP) activity and osteogenic markerexpression. The relationships among H19, miR-140-5p and SATB2 were examined through bioinformaticsanalyses, luciferase reporter assay, RIP assay and RNA pull-down assay. H19 expression was remarkablyincreased and miR-140-5p expression was dramatically reduced during osteogenic differentiation of BMMSCs. Functional analyses revealed that H19 overexpression or miR-140-5p depletion accelerated osteogenicdifferentiation of BM-MSCs. Conversely, H19 loss or miR-140-5p increase suppressed osteogenic differentiation of BM-MSCs. MiR-140-5p was confirmed as a target of H19, and miR-140-5p could bind to SATB2 aswell. Moreover, H19 knockdown reduced SATB2 expression by upregulating miR-140-5p. Additionally, miR140-5p depletion antagonized the inhibitory effect of H19 knockdown on osteogenic differentiation of BMMSCs. And, miR-140-5p inhibited osteogenic differentiation of BM-MSCs by targeting SATB2. In conclusion,H19 promoted osteogenic differentiation of BM-MSCs through regulating miR-140-5p/SATB2 axis, deepeningour understanding on the molecular mechanisms of H19 in coordinating osteogenesis.
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Objective: To summarize the effect of cartilage progenitor cells (CPCs) and microRNA-140 (miR-140) on the repair of osteoarthritic cartilage injury, and analyze their clinical prospects. Methods: The recent researches regarding the CPCs, miR-140, and repair of cartilage in osteoarthritis (OA) disease were extensively reviewed and summarized. Results: CPCs possess the characteristics of self-proliferation, expression of stem cell markers, and multi-lineage differentiation potential, and their chondrogenic ability is superior to other tissues-derived mesenchymal stem cells. CPCs are closely related to the development of OA, but the autonomic activation and chondrogenic ability of CPCs around the osteoarthritic cartilage lesion cannot meet the requirements of complete cartilage repair. miR-140 specifically express in cartilage, and has the potential to activate CPCs by inhibiting key molecules of Notch signaling pathway and enhance its chondrogenic ability, thus promoting the repair of osteoarthritic cartilage injury. Intra-articular delivery of drugs is one of the main methods of OA treatment, although intra-articular injection of miR-140 has a significant inhibitory effect on cartilage degeneration in rats, it also exhibit some limitations such as non-targeted aggregation, low bioavailability, and rapid clearance. So it is a good application prospect to construct a carrier with good safety, cartilage targeting, and high-efficiency for miR-140 based on articular cartilage characteristics. In addition, CPCs are mainly dispersed in the cartilage surface, while OA cartilage injury also begins from this layer, it is therefore essential to emphasize early intervention of OA. Conclusion: miR-140 has the potential to activate CPCs and promote the repair of cartilage injury in early OA, and it is of great clinical significance to further explore the role of miR-140 in OA etiology and to develop new OA treatment strategies based on miR-140.
ABSTRACT
Osteoarthritis is a kind of joint disease with cartilage injury as the main pathological changes.There is no effective treatment for them.With the discovery,research and application of microRNA (miR),it also provides new hope for the treatment of osteoarthropathy.At present,the role of miR in the pathogenesis of cartilage injury has been fully recognized.In this paper,we reviewed the role of miR-140 in the development of osteoarthritis and its important target genes in recent years.
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AIM:To explore the expression level of microRNA-140 ( miR-140 ) in human gastric cancer and normal gastric tissues, and the regulatory effect of miR-140 expression on the function of SGC-7901 cells.METHODS:The expression levels of miR-140 in human gastric cancer and normal gastric tissues were detected by real-time PCR.miR-140 mimics ( miR-140 up-regulated expression) and miR-140 inhibitors ( miR-140 down-regulated expression) were trans-fected into human gastric cancer SGC-7901 cells by liposome method.At the same time, the untransfected control group ( control group) and miRNA nonsense sequence transfection group ( NC group) were set up .The expression of miR-140 in the cells after transfection was detected by real-time PCR.The cell viability and growth inhibition rate with DDP were meas-ured by MTT assay.The cell cycle and apoptotic rate of SGC-7901 cells were analyzed by flow cytometry.The invasion a-bility of SGC-7901 cells was measured by Transwell assay.The protein expression of histone deacetylase 4(HDAC4) in the cells was determined by Western blot.RESULTS:The expression level of miR-140 in human gastric cancer tissues was significantly lower than that in normal gastric tissues (P<0.05).Compared with control group and NC group, the viability and invasion ability of the SGC-7901 cells were decreased, the cell cycle was arrested, the cell growth inhibition rate and apoptotic rate with DDP treatment were increased, and the protein expression of HDAC4 was down-regulated ( P<0.05) in miR-140 mimics group.However, in miR-140 inhibitors group, the viability and invasion ability of the SGC-7901 cells were increased, the cell cycle was promoted, the cell growth inhibition rate and apoptotic rate with DDP treatment were de-creased, and the protein expression of HDAC4 was up-regulated ( P<0.05 ) .CONCLUSION:The expression level of miR-140 in the gastric cancer tissues is low.miR-140 serves as a tumor suppressor to regulate the viability, apoptosis and invasion ability of gastric cancer cells, and to play a role by down-regulating HDAC4 protein.miR-140 may serve as a new target for diagnosis and treatment of gastric cancer.