miR-940 modulates CD47 to suppress biological functions of lung adenocarcinoma cells.

OBJECTIVE: mir-940 and CD47 play regulatory and immunoregulatory roles in lung cancer. While previous study found that the expression of mir-940 decreased, associated with the increasing of CD47 in lung adenocarcinoma. However, their inherent correlations remain elusive. Herein, this experiment intends to search for the relevant molecular mechanisms regulating the biological function of non-small cell lung cancer. METHODS: The cancer and adjacent tissue samples were collected from 20 pairs of newly diagnosed non-small cell lung cancer patients without applying radiotherapy and chemotherapy. We performed immunohistochemistry containing 45 lung adenocarcinoma tissues to investigate the relationship between the clinicopathological features and CD47 expression. The expressions of mir-940 and CD47 were detected by real-time quantitative polymerase chain reaction (qRT-PCR). Lung epithelial and lung adenocarcinoma (A549, H1299, GLC-82, PC-9) cell lines were cultured to detect the expression of mir-940 and CD47 molecules in each cell line. According to the expression situation, 2 cell lines were selected for mimic and siRNA transfection, and the transfection efficiency was also verified by qRT-PCR and western blot. CCK-8, transwell migration, transwell invasion, and colony formation assays were used to detect the changes in biological functions of lung adenocarcinoma cells after transfection, such as enhanced proliferation, migration, invasion, and cloning. The changes of related protein molecules after transfection were detected by western blot. The dual-luciferase experiment verified the targeting regulation relationship between mir-940 and CD47. Finally, flow cytometry analysis of apoptosis and cell cycle were carried out to detect apoptosis cells and change phase of cell cycle distribution. RESULTS: CD47 expression was not associated with clinicopathologic factors in lung adenocarcinoma. The proliferation, migration, invasion, and cloning abilities of lung adenocarcinoma cells were weakened after transfection with mir-940 mimic and siRNA-CD47. Overexpression of CD47 could promote proliferation, migration, invasion, cloning abilities, reduce apoptosis rate and attenuate the antitumor effect of mir-940 on lung adenocarcinoma. Dual luciferase experiments confirmed that mir-940 can target CD47 molecules. CONCLUSION: mir-940 can inhibit the biological function of lung adenocarcinoma cells by targeting CD47.

Metastasis-related long non-coding RNAs AL359220.1, SH3BP5-AS1 and ZF-AS1 are significant for prognostic assessment of lung adenocarcinoma.

BACKGROUND: Metastasis of lung adenocarcinoma (LUAD) severely worsens prognosis. Genetic alteration in the tumor microenvironment (TME) is closely associated with metastasis and other malignant biological properties of LUAD. In this study, we establish a metastasis-related risk model to accurately predict LUAD prognosis. METHODS: RNA-sequencing profiles and clinical data of LUAD patients including 503 tumor tissues and 54 adjacent normal tissues were collected in TCGA database. Additionally, the paired specimens from 156 LUAD patients were obtained in a single center. The metastatic relevance and clinical significance of metastasis-related long non-coding RNA (MRLNRs) was validated by series of in vitro experiments including western blotting, qPCR and transwell assays. RESULTS: Six MRLNRs were significantly correlated to prognoses of LUAD patients, of which AL359220.1, SH3BP5-AS1 and ZF-AS1 were further used to establish a metastasis-related risk scoring model (MRRS) due to the close associations with overall survival of LUAD patients. According to the MRRS, patients with higher scores in the high-risk group obtained poorer prognoses and survival outcomes. ZFAS1 expressed highly in tumor tissues and showed the inverse results compared to SH3BP5-AS1 and AL359220.1. In addition, the high expression of ZFAS1 was prominently correlated to the more advanced T-stage and distant metastasis. The reduction of ZFAS1 induced by siRNAs dramatically diminished the migration and invasion abilities of LUAD cells. CONCLUSIONS: In the present research, we elucidate the metastatic relevance and clinical significance of AL359220.1, SH3BP5-AS1 and ZF-AS1 in LUAD. Moreover, MRRS provide a promising assessing model for clinical decision making and prognosis of LUAD.

The role of miR-21/RECK in the inhibition of osteosarcoma by curcumin.

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.

[Effects of LncRNA UNC5B-AS1 on adhesion, invasion and migration of lung cancer cells and its mechanism].

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 (P＜0.05). The expression of UNC5B-AS1 in lung cancer A549 cells was the highest (P＜0.05). Down-regulation of UNC5B-AS1 expression inhibited adhesion, invasion and migration of A549 cells (P＜0.05). 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.