Aim To investigate the molecular mechanism of miR-326 inhibiting breast cancer invasion and metastasis by regulating EphB3 expression.Methods RTFQ-PCR was used to examine the expression o miR-326 in normal breast epithelial cells and breas cancer cells and the transfection efficiency of miR-326overexpression plasmid.EdU cell proliferation assay and Transwell assay were used to examine the changes in proliferation,migration and invasion ability of different subgroups of cells.Dual luciferase assay was used to verify the presence of binding sites for miR-326 and EphB3.Western blot was used to detect the expression of EphB3 in breast cancer cells after overexpression of miR-326.Results RTFQ-PCR results showed that miR-326 was lowly expressed in breast cancer cells and successfully transfected （P＜0.05）.EdU proliferation assay and Transwell assay results showed that overexpression of miR-326 in breast cancer cells inhibited proliferation,migration and invasive ability （P＜0.05）.The results of dual luciferase assay showed that miR-326 could interact with the 3’-UTR of EphB3 （P＜0.05）.Western blot and Transwell assays showed that miR-326 could negatively regulate EphB3 to inhibit invasive metastasis of breast cancer cells （P＜0.05）.Conclusions MiR-326 acts as a cancer suppressor genes in the development of breast cancer and suppresses the invasion and metastasis of breast cancer cells by regulating the expression of EphB3.

BACKGROUND:Lung adenocarcinoma (LUAD) is the most common clinical histological subtype of lung cancer and microRNAs (miRNAs) are a type of small non-coding RNAs which play a central role in cells. miR-30b-3p plays a key effect in many types of carcinoma, but there is still very little research on how it works in lung adenocarcinoma. The role and mechanism of miR-30b-3p in the proliferation and invasion of LUAD were explored in this study, to provide new targets for inhibiting the proliferation and invasion of LUAD.METHODS:NCBI database was used to screen out miRNA with obvious differential expression, and the differential expression and survival curve were searched by StarBase database. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the relative expression of miR-30b-3p in each lung adenocarcinoma cell line. 5-ethynyl-2'-deoxyuridine (EdU) cell proliferation assay and Transwell invasion assay were used to detect the proliferation and invasion of A549 cells in each group. The target genes of miR-30b-3p were determined by the target gene prediction websites. Western blot assay was used to detect the expression of COX6B1 in each group of A549 cells. Double luciferase assay was used to verify the targeted binding relationship between miR-30b-3p and COX6B1.RESULTS:The expression of miR-30b-3p in lung adenocarcinoma tissues and lung adenocarcinoma cells was downregulated (P<0.05). Low expression levels of miR-30b-3p were associated with poor prognosis in patients with lung adenocarcinoma (P=0.005,8). Overexpression of miR-30b-3p could inhibit the proliferation and the invasion of lung adenocarcinoma cells (P<0.05). Double luciferase assay proved that miR-30b-3p could target and bind to COX6B1 (P<0.05). Western blot analysis showed that the overexpression of miR-30b-3p could downregulate the expression of COX6B1 in A549 cells (P<0.05). EdU cell proliferation assay and Transwell invasion assay showed that the overexpression of miR-30b-3p could reverse the promoting effect of upregulation of COX6B1 on proliferation and invasion in lung adenocarcinoma cells (P<0.05).CONCLUSIONS:miR-30b-3p acts as a tumor suppressor gene in lung adenocarcinoma, and it can inhibit the proliferation and invasion of lung adenocarcinoma by targeting the expression of COX6B1.

The roles of microRNAs (miRNAs) in the occurrence, metastasis, and prognosis of lung adenocarcinoma (LUAD) have been drawing extensive attention from researchers. The aim of this study is to identify the effects of miR-4732-5p on the migration, invasion, and metastasis of LUAD. In this study, we found that the expression of miR-4732-5p was decreased in LUAD based on the data derived from The Cancer Genome Atlas (TCGA) database, tissues, and cell lines. LUAD patients with a low expression of miR-4732-5p exhibited a lower survival rate. Meanwhile, miR-4732-5p could directly target xenotropic and polytropic retrovirus receptor 1 (XPR1), and elevated XPR1 was observed in LUAD mRNA microarrays, Gene Expression Omnibus (GEO), and The Human Protein Atlas (HPA) database. Overexpression of miR-4732-5p significantly inhibits the migration, invasion, and metastasis of LUAD in vitro and in vivo, which can be reversed by overexpression of XPR1. We also found that the PI3K/Akt/GSK3β/Snail pathway induced by EGF induced EMT could be inhibited by miR-4732-5p overexpression and XPR1 knockdown. The migration and invasion of LUAD could be converted by cytoskeletal rearrangements, and the polymerization of EGF induced F-actin in A549 cells could be inhibited by elevated miR-4732-5p. Our results suggest that miR-4732-5p exerts anti-tumor effects on the invasion and metastasis of LUAD by regulating XPR1 in vivo and in vitro, indicating that the miR-4732-5p/XPR1 axis may be a potential target for LUAD therapeutic intervention.

在新医科大力发展的背景下,教师在讲授肿瘤病理学专业知识的同时,如何挖掘肿瘤病理学的思政元素,将知识传授与价值引领结合起来,如何达到既能开展课程思政,又能培养德才兼备的医学人才是教学过程中应该思考的问题.要结合临床病例将思政教育融入肿瘤病理学课程,帮助医学研究生提高人文素养.

Background Long non-coding RNAs (LncRNAs) has been confirmed to play a crucial role in the development and progression of various cancer types. Here we evaluated the expression profiles of LncRNAs in Lung adenocarcinoma (LUAD) tissues and identified a novel LncRNA, termed LncRNA-AC009948.5. However, the role and potential molecular mechanisms of this novel LncRNA in LUAD carcinogenesis is unknown. Methods Regarding the public databases and based on integrating bioinformatics analyses, we determined whether LncRNA-AC009948.5 exerts its oncogenic functions via sponging miR-186-5p in LUAD. Furthermore, we determined whether NCAPG2 was a downstream target of miR-186-5p. Moreover, the expression level and biological function of LncRNA-AC009948.5 in LUAD were determined by qRT-PCR, cell apoptosis, Edu, transwell, wound healing and western blot assays. Besides, xenograft mice were established for validation. We explored the expression of LncRNA-AC009948.5 and its roles in the prognosis of LUAD. Results LncRNA expression microarray data indicate that LncRNA-AC009948.5 is upregulated in LUAD samples. The present study confirmed the upregulation of LncRNA-AC009948.5 in LUAD tissues and cells. Encreased expression of LncRNA-AC009948.5 was correlated with tumor size, lymph nodes, distant metastasis and histological grade, and poor prognosis.LncRNA-AC009948.5 knockdown significantly inhibited cell proliferation, migration, and invasion in vitro , as well as tumorigenesis and metastasis in vivo . Conversely, LncRNA-AC009948.5 upregulated had opposite effects. Mechanistically, we elucidated that LncRNA-AC009948.5 could directly bind to miR-186-5p and subsequently suppress expression of the target gene of NCAPG2. Conclusions LncRNA-AC009948.5 promotes lung adenocarcinoma cells metastasis via the miR-186-5p/NCAPG2 axis and activation of the EMT process. Which may serve as potential targets for the treatment of LUAD in the future.

Abstract Background: Glioma is one of the most common intracranial malignancies. As competing for endogenous RNA (ceRNA), long chain non-coding RNA (lncRNA) can affect the binding of microRNA (miRNA) to mRNA and thus affect the occurrence and development of glioma, which has been suggested as a key role in the development of human cancers. This study was to investigate the effects of TCGA on the development of glioma.Results: Here, we screened the differential genes by differential analyses on the matrix data for glioma in TCGA and GEO databases and selected the potential prognostic markers among them. Surprisingly, LEF1-AS1 was identified capable of influencing the survival of glioma patients. The results derived from the proportional risk (Cox) regression model showed a significant association between LEF1-AS1 overexpression and poorer prognosis (HR = 1.518, P < 0.001). The ROC model validated the feasibility of LEF1-AS1 (AUC = 0.702). MiR-128-3p, the target gene for LEF1-AS1, was screened using the Bioinformatics Online website and found down-regulated with increasing glioma grade in glioma tissues. Then the target gene for miR-128-3p, CSRP2, was obtained through miRTarBase, TargetScan, and miRDB. The survival curve of CSRP2 showed that CSRP2 with low expression corresponded to a better prognosis in glioma patients. The level of CSRP2 was positively correlated with the infiltration ones of CD4+ T and CD8+ T cells in glioma tissues. In addition, the CellMiner database predicted three drugs (Elesclomol, Entinostat, and Staurosporine) with high sensitivity to CSRP2 expression, and their molecular structures were retrieved using the PubChem22 database.Conclusions: In summary, the LEF1-AS1/miR-128-3p/CSRP2 axis may play a vital role in the development of glioma and provide guiding significance for the early diagnosis and treatment for it.

目的 分析分泌型卷曲相关蛋白4(SFRP4)在胃癌细胞中的表达,探讨SFRP4对胃癌细胞迁移以及侵袭的影响机制.方法 选取正常胃黏膜上皮细胞GES-1和胃癌细胞SGC-7901分别培养,蛋白质印迹法检测SFRP4表达.构建SFRP4对照质粒及敲低质粒转染至SGC-7901细胞,分别作为Scr组和SiSFRP4组,另外设置空白组(SGC-7901),通过蛋白质印迹法检测质粒转染效率;划痕实验以及Transwell侵袭实验分别检测转染质粒后胃癌细胞的迁移以及侵袭能力;蛋白质印迹法检测转染质粒后胃癌细胞中基态金属蛋白酶(MMPs)和上皮间充质转化(EMT)标志物的表达变化.结果 胃癌细胞株SGC-7901 SFRP4蛋白表达水平为2.54±0.19,高于GES-1细胞(1.00±0.11),差异有统计学意义,t=12.150,P<0.001.质粒转染后,Scr组SFRP4表达水平为0.95±0.10,SiSFRP4组为0.27±0.06,差异有统计学意义,t=10.100,P<0.001.SiSFRP4组穿过Transwell小室Matrigel滤膜的细胞数量(0.41±0.10)低于Scr组(1.00±0.12),t=6.542,P=0.003.转染24和48 h后,SiSFRP4组细胞迁移率分别为0.42±0.13和0.41±0.08,低于Scr组的1.00±0.17和1.00±0.08,差异有统计学意义,t值分别为4.693和8.851,P值分别为0.009和<0.001.转染24 h后,SiSFRP4组E-cadherin表达(2.35±0.15)高于Scr组(1.00±0.22),差异有统计学意义,t=8.782,P<0.001;SiS-FRP4组Vimentin表达(0.31±0.14)低于Scr组(1.00±0.27),差异有统计学意义,t=3.930,P=0.017;SiSFRP4组MMP-2及MMP-9表达水平分别为0.58±0.09和0.66±0.08,低于Scr组的1.00±0.12和1.00±0.10,差异有统计学意义,t值分别为4.850和4.599,P值分别为0.008和0.010.结论 SFRP4在胃癌细胞中高表达,且可通过EMT途径及上调MMPs表达促进胃癌细胞的迁移以及侵袭过程.

BACKGROUND:MicroRNAs (miRNAs) are short non-coding RNAs that regulate gene expression, influence cellular processes, and promote disease development. Variations in miRNA expression have been observed in many diseases, including hepatitis, cardiovascular disease, and cancer. The aim of this study is to investigate the effect of miR-144-3p on the invasion and metastasis of lung adenocarcinoma by targeting recombinant insulin receptor substrate 1 (IRS1).METHODS:The expression of miR-144-3p in patients with lung adenocarcinoma was queried through bioinformatics database. MirTarPathway was used to analyze the KEGG enrichment pathway of miRNA. The expression and plasmid transfection efficiency of miR-144-3p in lung adenocarcinoma cell lines were detected by quantitative reverse transcription polymerase chain reaction (qRT-PCR). Transwell assay was used to detect the changes of cell invasion and migration ability in different groups. Bioinformatics determined the key genes (Hub genes) of miR-144-3p; Double luciferase target assay was used to detect the mutual binding of miR-144 and IRS1. Western blot assay was used to detect the expression of IRS1 in different cell lines and the expression of after overexpression of miR-144.RESULTS:The expression of miR-144-3p in lung adenocarcinoma tissues was decreased, qRT-PCR results indicated that the expression of miR-144-3p in lung adenocarcinoma cell A549 was significantly decreased (P<0.05), and the overexpressed plasmid was successfully transfected (P<0.05). Overexpression of miR-144 decreased the ability of cell migration and invasion (P<0.05). The expression of IRS1 was up-regulated in lung adenocarcinoma tissues. Survival analysis showed that patients with lung adenocarcinoma with high IRS1 expression had a poor prognosis (P<0.05). Double luciferase assay results showed that miR-144 could specifically identify 3'-UTR of IRS1 and inhibit reporter enzyme expression (P<0.05). Western blot indicated that the expression of IRS1 was increased in A549 cells (P<0.05). After overexpression of miR-144, the expression level of IRS1 protein was decreased (P<0.05). Transwell experiment proved that miR-144-3p could inhibit invasion and metastasis of lung adenocarcinoma cells by targeting IRS1 (P<0.05).CONCLUSIONS:MiR-144-3p inhibits the invasion and migration of A549 cells through targeted regulation of IRS1, thus playing an anticancer role in tumors.

目的 本研究通过小鼠急性氯化汞(Mercury dichloride,HgCl2)中毒模型,研究肾水通道蛋白-1(Aquaporin-1,Aqp1)及mmu-miR-429-5p的表达变化,从而探究其可能发挥作用的分子机制,为急性HgCl2中毒后的临床治疗提供更多的靶点以及为司法鉴定提供更多思路.方法 全自动生化分析仪测定肾功能指标水平,苏木素-伊红(HE)染色观察肾组织结构病理改变.免疫组织化学实验及Western blot检测Aqp1蛋白定量及定位.生物信息学网站预测能与Aqp1靶向结合的微小RNA(MicroRNAS,miRNAS).结合双荧光素酶报告基因实验检测mmu-miR-429-5p与Aqp1 之间是否存在结合靶点,实时荧光定量 PCR(quantitative Real-Time PCR,qRT-PCR)检测 Aqp1 及 mmu-miR-429-5p在肾组织中的表达水平.结果 急性HgCl2中毒后肾小管上皮细胞水肿及变性坏死,Aqp1在肾小管上皮细胞膜、肾小球毛细血管内皮细胞膜上表达降低.双荧光素酶报告基因实验结果显示mmu-miR-429-5p与Aqp1之间存在靶点结合.QRT-PCR结果显示Aqp1基因表达降低而mmu-miR-429-5p表达升高,两者之间存在负相关关系.结论 急性HgCl2中毒后mmu-miR-429-5p表达升高并可能通过靶向结合Aqp1致Aqp1蛋白表达降低,从而引起肾损伤.

Background and purpose: Lung adenocarcinoma is a subtype of non-small cell lung cancer. Although much progress has been made in the diagnosis and treatment of lung adenocarcinoma, the clinical prognosis and overall survival of advanced lung adenocarcinoma are still poor. In recent years, a number of studies have shown that miRNA can play a role in a variety of cancers, and play an important role in cell proliferation, metastasis, inflammation and other biological processes. This study aimed to explore the effect of miR-625-5p on the proliferation and invasion ability of lung adenocarcinoma cells and its molecular mechanism, so as to provide a new idea for the diagnosis and treatment of lung cancer. Methods: GEO database was used to search for differentially expressed miRNA in lung adenocarcinoma. Real-time fluorescence quantitative polymerase chain reaction (RTFQ-PCR) was used to detect the expression of miR-625-5p in various lung adenocarcinoma cell lines. The effects of miR-625-5p on the proliferation and invasion of lung adenocarcinoma cells were investigated by EdU cell proliferation assay and transwell invasion assay. Key genes of miR-625-5p targeted binding were predicted by bioinformatics. Western blot experiment validated the expression of protein kinase cAMP-activated catalytic subunit alpha (PRKACA) in lung adenocarcinoma cells. The targeted relationship between miR-625-5p and PRKACA was analyzed by double luciferase assay and Western blot. Western blot assay was used to detect the expression of PRKACA after co-transfection of si-PRKACA and si-miR-625-5p in each group. The effect of miR-625-5p on the proliferation and invasion ability of lung adenocarcinoma cells by targeting PRKACA was observed by EdU cell proliferation assay and transwell invasion assay. Results: The expression of miR-625-5p was up-regulated in lung adenocarcinoma tissues (P<0.000 1) and cells (P<0.000 1). The results of EdU cell proliferation assay and transwell invasion assay showed that miR-625-5p promoted the proliferation (P=0.002 3) and invasion (P=0.000 3) of lung adenocarcinoma cells. Double luciferase assay showed that miR-625-5p could target and bind to PRKACA (P=0.000 8). In lung adenocarcinoma cells, miR-625-5p was negatively correlated with PRKACA expression (P<0.000 1). Down-regulation of miR-625-5p reversed the promotion of the proliferation (P=0.011 9) and invasion (P=0.001 5) ability of A549 cells by knockout of PRKACA. Conclusion: MiR-625-5p is up-regulated in lung adenocarcinoma and promotes proliferation and invasion of lung adenocarcinoma tissues and cells by negatively regulating PRKACA.

OBJECTIVETo explore the role of miR-671-5p in regulating the migration and invasion of osteosarcoma and the underlying mechanisms.METHODSThe differentially expressed microRNAs (miRNAs) in osteosarcoma were screened in the NCBI online database, and the target proteins of these miRNAs were predicted and their functions were analyzed. Osteosarcoma cells were transfected with a plasmid overexpressing miR-671-5p, and the transfection efficiency was assessed using quantitative real-time PCR (qRT-PCR). The changes in the migration and invasion of the transfected cells were examined with Transwell assay, and the expressions of proteins related with epithelial-mesenchymal transition (EMT) were detected using Western blotting. Dual-luciferase reporter assay was performed to determine whether the 3'UTR of SMAD3 contained a targeted binding site of miR-671-5p.RESULTSMiR-671-5p was significantly down-regulated in both osteosarcoma tissues and osteosarcoma cells (P < 0.05). The osteosarcoma cells overexpressing miR-671-5p showed significantly reduced migration and invasion abilities (P < 0.05) with obviously lowered expressions of EMT-related proteins (P < 0.05). SMAD3 was highly expressed in osteosarcoma cells (P < 0.05), and dual-luciferase reporter assay confirmed the presence of a targeted binding site between miR-671-5p and the 3'UTR of SMAD3 (P < 0.05). In osteosarcoma cells transfected with a SMAD3-overexpressing plasmid (P < 0.05), the high expression of SMAD3 significantly inhibited by miR-671-5p overexpression (P < 0.05). Transwell assay demonstrated that SMAD3 overexpression significantly promoted the migration and invasion of osteosarcoma cells (P < 0.05), and while miR-671-5p overexpression obviously reversed this effect (P < 0.05).CONCLUSIONMiR-671-5p can inhibit the invasion and migration of osteosarcoma cells by negatively regulating SMAD3.

已有研究证实,microRNA (miRNAs)通过调控多种基因参与肿瘤的发生和发展过程.然而,miR-654-5p通过调控宿主细胞因子1(host cell factor 1,HCFC1)抑制乳腺癌细胞侵袭转移的分子机制尚不清楚.本研究通过生物信息学数据库分析发现,miR-654-5p在乳腺癌组织中低表达,且低表达病人愈后较差(P=0.013),实时荧光定量PCR(quantitative real-time PCR,qRT-PCR)检测表明,miR-654-5p在MDA-MB-231细胞中表达降低(P＜0.05),且与对照组相比,转染过表达质粒后,miR-654-5p的表达明显升高(P＜0.05).5-乙炔基-2脱氧尿嘧啶核苷(EdU)增殖实验和Transwell实验表明,过表达miR-654-5p能够抑制MDA-MB-231细胞的增殖、迁移和侵袭能力(P＜0.05).运用Cytoscape软件筛选出miR-654-5p的Hub基因HCFC1,发现miR-654-5p与HCFC1的表达呈负相关,且HCFC1在乳腺癌组织中的表达水平较高并与淋巴结转移密切相关,此外,HCFC1高表达病人的愈后较差(P=0.0039).双荧光素酶实验证实,miR-654-5p能够与HCFC1 mRNA的3 '-UTR结合(P＜0.05).Western印迹结果显示,与人正常乳腺上皮细胞MCF-10A相比,HCFC1在MDA-MB-231细胞中表达水平较高(P＜0.05),过表达miR-654-5p后HCFC1的表达显著下调(P＜0.05).Transwell实验结果显示,与对照组相比,过表达miR-654-5p后MDA-MB-231细胞的迁移和侵袭能力明显下降,而共转HCFC1在一定程度上能够逆转miR-654-5p对于乳腺癌细胞迁移和侵袭能力的抑制作用(P＜0.05).综上所述,miR-654-5p可通过调节HCFC1的表达进而抑制乳腺癌细胞的增殖、侵袭和转移.