miR-216b-5p Promotes the Migration and Invasion of LN-229 Glioma Cells by Targeting BTN3A2 / 中国生物化学与分子生物学报

Accumulating evidence indicated that microRNAs (miRNA) play an important role in tumor invasion and metastasis by regulating their target genes.However, whether the miRNA-216b-5p(miR-216b-5p ) and their target genes butyrophilin subfamily 3 member A2(BTN3A2) promote glioma invasion and metastasis is unclear.This study aims to study whether miR-216b-5p promoted migration and invasion in glioma cells by negatively regulating BTN3A2.The differential expression analysis of GSE15824 and GSE4290 was analyzed by GEO2R.We found that only BTN3A2 is up-regulated in both GSE15824 and GSE4290 (P<0.05).The gene set enrichment analysis (GSEA) analysis indicated BTN3A2 was related to many cancer-related pathways (P<0.05).The results of survival curves showed that the overall survival of patients with high expression of BTN3A2 decreased significantly (P <0.001).The expression of BTN3A2 was increased with the increase of WHO grade (P<0.05), while the expression of BTN3A2 was increased in 1p/19q uncombined deletion and IDH mutant patients (P<0.001).Western blotting results showed that BTN3A2 was up-regulated in seven glioma tissues and glioma cell lines U87, U251 and LN-229 and downregulated in the miR-216b-5p mimics group; Transwell results showed that transfection with BTN3A2 silencing plasmids(si-BTN3A2) or miR-216b-5p mimics plasmids could inhibit the ability of migration and invasion in LN-229 cells in vitro (P<0.05).The online websites predicted miR-216b-5p as a potential target gene of BTN3A2.The survival curve results show that compared with patients with low expression of miR-216b-5p , the survival rate of patients with high expression was significantly increased (P=0.025).The relative expression of miR-216b-5p was decreased in U87, U251 and LN229 cells was detected by real time quantitative PCR (P<0.05).The results of dual luciferase assay showed that BTN3A2 could bind to miR-216b-5p (P<0.05).Transwell experiment results showed that overexpression of miR-216b-5p can inhibit the migration and invasion ability of LN229 cells (P<0.05).In summary, miR-216b-5p promotes the migration and invasion by targeting BTN3A2 of LN-229 glioma cells.

MiR-654-5p Inhibited Invasion and Metastasis of Breast Cancer Cells by HCFC1 / 中国生物化学与分子生物学报

Studies have confirmed that microRNA (miRNAs) is involved in the development and progression of tumors by targeting multiple genes. However, the molecular mechanism of miR-654-5p in in- hibiting the invasion and metastasis of breast cancer cells through the targeted regulation of host cell factor 1 (HCFCl) is still unclear. The analysis of bioinformatics datasets found that miR-654-5p was downregu-lated in breast cancer tissues and was associated with poor prognosis (P = 0. 013). Quantitative real-time PCR (Quantitative real-time PCR, qRT-PCR) showed that the expression of miR-654-5p in MDA-MB-231 cells was decreased (P<0. 05), and the expression of miR-654-5p was significantly increased after transfection of the overexpressed plasmid (P<0. 05) as compared with the control group. The 5-Ethynyl-2'-deoxyuridine (EdU) proliferation experiment and Transwell assay showed that overexpression of miR-654-5p inhibited the proliferation, migration and invasion of MDA-MB-231 cells (P<0. 05). Hub gene HCFCl of miR-654-5p was screened and constructed by Cytoscape software, and it was found that miR-654-5p was negatively correlated with the expression of HCFCl. The expression of HCFCl was increased in breast cancer tissues and closely correlated with lymph node metastasis, and patients with high expression of HCFCl had poor prognosis (P = 0. 0039). Dual-luciferase assay confirmed that miR-654-5p could bind to the 3'-UTR of HCFClmKNA (P<0. 05). Western blot results showed that compared with human normal breast epithelial cells MCF-10A, the expression of HCFCl was increased in MDA-MB-231 cells (P<0. 05), and the expression of HCFCl was significantly down-regulated after overexpression of miR-654-5p (P<0. 05). Transwell experiment results showed that the migration and invasion ability of MDA-MB-231 cells after overexpression of miR-654-5p was significantly decreased compared with the control group. Co-transfection of HCFCl can partially reverse the inhibitory effect of miR-654-5p on the migration and invasion ability of breast cancer cells (P<0. 05). In conclusion, miR-654-5p can inhibit the proliferation, invasion and metastasis of breast cancer cells by regulating the expression of HCFCl.

Bioinformatics analysis and experimental validation of microRNA-140-3p targeting cell division cycle associated 8 inhibiting invasion and metastasis of lung adenocarcinoma cells / 解剖学报

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.

Survey on WeChat Operation in Municipal Public Hospitals / 医学信息学杂志

The paper surveys on WeChat operation in 34 municipal public hospitals in Shanghai,including aspects like functional positioning,information display,service function and characteristic function and so on,and puts forward related suggestions on development of hospital official WeChat,including aspects like making industrial standard,promoting tiered diagnosis and treatment and giving importance to data collection,etc.

Effect of EphB4/EphrinB2 reverse signal on angiogenesis induced by Xuefu Zhuyu Capsule () containing serum in human microvascular endothelial cell 1 / 中国结合医学杂志

<p><b>OBJECTIVE</b>To evaluate the effect of Xuefu Zhuyu Capsule ()-containing serum (XFZY-CS) on EphB4/ephrinB2 and its reverse signal in human microvascular endothelial cell-1 (HMEC-1).</p><p><b>METHODS</b>XFZY-CS and the blank control serum were collected. HMEC-1 cells were randomly assigned to 6 groups including the concentration 1.25%, 2.5%, and 5% XFZY-CS groups and their blank serum control ones. The angiogenesis effect of XFZY-CS was tested with an in vitro tube formation assay and the best condition of pro-angiogenesis was determined. The effect of XFZY-CS on EphB4/ephrinB2 and the reverse signal were determined by Western blot and real-time quantitative polymerase chain reaction, respectively; we also confifirmed the results through activating and inhibiting the reverse signal by EphB4/fc and pyrophosphatase/ phosphodiesterase2 (PP2).</p><p><b>RESULTS</b>XFZY-CS promoted angiogenesis at the concentration of 2.5% corresponding serum after being cultured for 48 h, while inhibited angiogenesis at the concentration of 5% after culturing for 48 and 72 h. Under the 2.5% serum concentration, XFZY up-regulated the expression of EphB4-mRNA at 12 h (P<0.05), and down-regulates its expression at 24 h (P<0.01). Protein expression of EphB4 was apparently up-regulated at 12 h and down-regulated at 24 h. The phosphorylation of ephrinB2 increased at 9 h (P<0.05). In addition, 2.5% XFZY-CS played a similar role as the reverse signaling activator EphB4/Fc ranging from 0.5 to 5 μg/mL (P>0.05). XFZY-CS also reduced the inhibitive effect of PP2 in limited periods.</p><p><b>CONCLUSIONS</b>EphB4/ephrinB2 was the upstream signal in the process of angiogenesis and its reverse signaling was responsible for XFZY's effect on promoting angiogenesis.</p>

Roles of bFGF in endothelial cell tube formation induced by xuefu zhuyu decoction under non-anoxia condition / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To explore the roles of basic fibroblast growth factor (bFGF) on tube formation induced by xuefu zhuyu decoction (XZD) under non-anoxia condition.</p><p><b>METHODS</b>Using serum pharmacology technique, endothelial cell line ECV304 cells were incubated in routine 95% O2. ECV304 cells were intervened by 1.25%, 2.50%, and 5.00% XZD containing serums and the vehicle serum for 48 h. The effects of XZD on tube formation, bFGF contents and its transcription levels were assessed by in vitro tube formation assay, enzyme-linked immunosorbent assay (ELISA), and reverse transcriptase polymerase chain reaction (RT-PCR), respectively.</p><p><b>RESULTS</b>Three concentrations of XZD containing serums could not only obviously promote the tube formation bFGF level, but also up-regulate bFGF contents in the supernate and its transcription levels. The shapes of lumens were more regular in those induced by 1.25% and 2.50% XZD containing serums.</p><p><b>CONCLUSION</b>XZD induced angiogenesis via up-regulating the bFGF expression under non-anoxia condition.</p>