ABSTRACT
Osteosarcoma is a malignant tumor in bones that is common in children and adolescents. MicroRNAs (miRs) are small non-coding RNAs that are associated with various kinds of tumors. miR-21 is one of the most frequently overexpressed microRNAs in osteosarcoma. Curcumin is a naturally occurring phenolic compound that has antitumor properties. Curcumin significantly inhibits osteosarcoma. However, the role of miR-21 and its target gene, reversion-inducing cysteine-rich protein with kazal motifs (RECK), in the anticancer activity of curcumin against osteosarcoma remains unclear. The aim of this study is to investigate the effect(s) of curcumin on osteosarcoma cell proliferation and elucidate its molecular mechanism. Cell counting kit-8, colony formation and flow cytometry assays were performed to study cell proliferation and apoptosis. Real time-polymerase chain reaction was used to determine the expression of miR-21 and RECK. Wnt/ß-catenin signaling pathway proteins were detected by Western Blot. We hereby show that curcumin upregulated the expression of RECK via miR-21, thereby subsequently regulating Wnt/ß-catenin signaling leading to the inhibition of osteosarcoma.
Subject(s)
Antineoplastic Agents/pharmacology , Bone Neoplasms/metabolism , Curcumin/pharmacology , GPI-Linked Proteins/metabolism , MicroRNAs/metabolism , Osteosarcoma/metabolism , Wnt Signaling Pathway/drug effects , Apoptosis/drug effects , Apoptosis/genetics , Bone Neoplasms/genetics , Cell Line, Tumor , Cell Proliferation/drug effects , Cell Proliferation/genetics , GPI-Linked Proteins/genetics , Gene Expression Regulation, Neoplastic/drug effects , Gene Expression Regulation, Neoplastic/genetics , Humans , MicroRNAs/genetics , Osteosarcoma/genetics , Real-Time Polymerase Chain Reaction , Up-Regulation , Wnt Signaling Pathway/geneticsABSTRACT
Objective: To investigate the effects of long-chain non-coding RNA (lncRNA) UNC5B-AS1 on the adhesion, invasion and migration of lung cancer cells by regulating the expression of miR-218-5p. Methods: Twenty specimens of lung cancer patients and corresponding paracancerous tissues were surgically removed and collected from the oncology department of Chongqing Three Gorges Central Hospital from June 2017 to June 2019. Real-time quantitative PCR (qRT-PCR) was used to detect the expressions of UNC5B-AS1 in human bronchial epithelial cells HBE and different lung cancer cells of A549, H1437, H1975, H1299 and H460. UNC5B-AS1 siRNA was transfected into lung cancer A549 cells. Adhesion assay, transwell invasion assay and scratch assay were used to detect the effect of UNC5B-AS1 on adhesion, invasion and migration of A549 cells. qRT-PCR and dual luciferase reporter gene were used for the detection and identification of UNC5B-AS1 targeting miR-218-5p. The expression of epithelial-mesenchymal transition (EMT)-related protein was detected by Western blot. Results: The expression of UNC5B-AS1 in lung cancer tissues and cells was significantly higher than that in adjacent tissues and bronchial epithelial cells (Pï¼0.05). The expression of UNC5B-AS1 in lung cancer A549 cells was the highest (Pï¼0.05). Down-regulation of UNC5B-AS1 expression inhibited adhesion, invasion and migration of A549 cells (Pï¼0.05). qRT-PCR and dual luciferase reporter assay experiments showed that UNC5B-AS1 targeted the regulation of miR-218-5p expression. Down-regulation of UNC5B-AS1 inhibited E-cadherin protein expression and promoted Vimentin and Twist protein expression. Conclusion: lncRNA UNC5B-AS1 promotes adhesion, invasion and migration of lung cancer cells through targeted regulation of miR-218-5p expression, and its mechanism may be related to the promotion of EMT.
