miR-181a-5p和miR-451a在胃癌诊断和预后评估中的临床价值

目的 探讨miR-181a-5p和miR-451a在胃癌诊断和预后评估的临床价值.方法 选取2014年1月至2016年12月在该院就诊的胃癌患者血清标本117例为胃癌组,选择同期在该院行胃镜检查诊断为息肉的患者血清标本75例和健康体检者血清标本45例分别设为胃息肉组和健康对照组.比较各组研究对象血清miR-181a-5...     

C-erbB2、c-myc和CCND1 mRNA联合检测在乳腺癌诊断中的应用

目的建立荧光定量聚合酶链反应（FQ-PCR）法检测C-erbB2、核内原癌基因（c-myc）和细胞周期蛋白D1（CC-ND1）mRNA水平,探讨其联合检测在乳腺癌诊断和治疗监测中的应用。方法建立FQ-PCR法,并以β2-微球蛋白为内对照测定15名健康女性体检者（正常对照组）、30例良性乳腺疾病患者（良性乳腺疾病组）和121例乳腺癌患者（乳腺癌组）外周血中C-erbB2、c-myc和CCND1的mRNA水平。结果 C-erbB2、c-myc和CCND1的mRNA水平在正常对照组和良性乳腺疾病组间差异无统计学意义（P〉0.05）,乳腺癌组均高于前两组（P〈0.05）,β2-微球蛋白三组间差异无统计学意义（P〉0.05）;三种mRNA联合检测的阳性率高于单个mRNA的检测。结论 FQ-PCR技术可快速定量检测C-erbB2、c-myc和CCND1的mR-NA,三者联合检测可有效提高对乳腺癌诊断的阳性率。     

miR-892a regulated PPP2R2A expression and promoted cell proliferation of human colorectal cancer cells

Colorectal cancer (CRC) is one of the most common malignancy cancers in the world. Aberrant microRNA expression is involved in human diseases including cancer. In the present study, we investigated the miR-892a's role in CRC cell proliferation. We found that miR-892a was frequently upregulated in human colorectal cancer tissues and cell lines compared with the matched tumor adjacent tissues and normal colonic cell line FHC. Overexpression of miR-892a promoted cell proliferation and colony formation of CRC. Bioinformatics analysis further revealed PPP2R2A was identified as a potential miR-892a. Overexpression of miR-892a-in SW480 cells reduced PPP2R2A protein expression. Subsequently, data from luciferase reporter assays showed that PPP2R2A 3′-untranslated region (3′-UTR) carried the directly binding site of miR-892a. Furthermore, siRNA-mediated silencing of PPP2R2A blocked the inhibitory effect of miR-892a-in on CRC cell growth. In sum, our data provided compelling evidence that overexpression of miR-892a may provide a selective growth promotion for CRC cells by direct suppression of PPP2R2A expression.     

荧光定量聚合酶链反应检测CCND1基因扩增在乳腺癌诊断中的应用

目的应用荧光定量聚合酶链反应（FQ-PCR）法检测细胞周期蛋白D1（CCND1）基因表达水平,探讨其在乳腺癌诊断和治疗监测中的应用价值。方法应用FQ-PCR法,并以β2-微球蛋白为内对照测定15例健康女性体检者、30例良性乳腺疾病患者和81例乳腺癌患者外周血中CCND1的表达量。结果CCND1表达水平在健康对照组和良性乳腺疾病组间差异无统计学意义（P〉0．05）,乳腺癌组均高于前2组（P〈0．05）,β2-微球蛋白在3组间差异无统计学意义（P〉0．05）。81例乳腺癌患者阳性率为45．7％,良性乳腺疾病组为0。结论FQ-PCR技术是高度灵敏、高度特异的快速定量检测CCND1方法,可有效监测乳腺癌的诊断：疗效、转移,评估其预后。     

miR-601 is a prognostic marker and suppresses cell growth and invasion by targeting PTP4A1 in breast cancer

MicroRNAs (miRNA) play important roles in the initiation and progression of breast cancer. Here, we investigated the role of miR-601 in breast cancer and found that its expression was significantly down-regulated in breast cancer tissues compared with matched adjacent non-cancerous breast tissues. Moreover, we found that down-regulation of miR-601 was closely associated with distant metastasis and poor distant metastasis-free survival in breast cancer. In addition, miR-601 levels were inversely correlated with metastatic potential of human breast cancer cell lines. Further experiments showed that ectopic overexpression of miR-601 suppressed breast cancer cell proliferation, migration and invasion, whereas miR-601 knockdown promoted breast cancer cell proliferation, migration and invasion. Furthermore, protein tyrosine phosphatase type IVA 1 (PTP4A1) was identified as a direct target of miR-601. Overexpression of miR-601 repressed PTP4A1 mRNA and protein expression. Conversely, inhibition of miR-601 increased PTP4A1 mRNA and protein expression. Taken together, our data suggest that miR-601 inhibits growth and invasion of breast cancer cells by targeting PTP4A1 and that miR-601 is a potential biomarker for prognosis and therapeutic target in breast cancer.     

腹腔镜结直肠癌手术治疗对结直肠癌患者血清中miR-17、miR-21、miR-20a和miR-92水平的影响

目的探究腹腔镜结直肠癌手术对结直肠癌(CRC)患者血清miR-17、miR-21、miR-20a和miR-92水平的影响。方法选取2015年6月-2018年6月该院收治的CRC患者120例作为研究对象,采用实时荧光定量PCR反应(RT-qPCR)检测血清中miR-17、miR-21、miR-20a和miR-92水平;使用生活质量核心调查问卷第3版(QLQ-C30)评估腹腔镜结直肠癌手术治疗前后患者的状况。结果经腹腔镜结直肠癌手术治疗后,CRC患者的躯体、认知、情绪、角色和社会功能评分均得到明显提高;患者血清中miR-17、miR-21、miR-20a和miR-92的相对水平均明显下降;通过相关性分析发现,miR-17、miR-20a和miR-92的相对量变化与QLQ-C30评分变化有相关性(P<0.05),而miR-21则无相关性(P>0.05)。结论腹腔镜结直肠癌手术治疗后,CRC患者血清中miR-17、miR-20a和miR-92的水平可以作为评估治疗效果的潜在标志物。     

乳腺癌中miR-141表达与临床病理特征、Keap1/Nrf2通路及预后的关系

目的 研究乳腺癌中miR-141表达与临床病理特征、KELCH样ECH关联蛋白1(Keap1)/核因子E2相关因子2(Nrf2)通路及预后的关系.方法 选择2014年3月至2015年3月期间在本院接受手术治疗的乳腺癌患者作为研究对象,检测乳腺癌组织和癌旁组织中miR-141、Keap1、Nrf2的表达水平,随访乳腺癌患...     

miR-29a在乳腺癌组织中的表达及其对乳腺癌细胞增殖和迁移的影响

摘要：目的：检测miR-29a在乳腺癌组织中的表达水平并探讨其对人乳腺癌细胞增殖和迁移的影响。 方法：用荧光定量RT-PCR检测乳腺癌组织及癌旁组织中miR-29a的表达水平;用脂质体法将miR-29a mimic及miR-29a inhibitor分别转染乳腺癌细胞系MDA-MB-231,通过MTT实验和 Transwell实验观察miR-29a对MDA-MB-231细胞增殖和迁移能力的影响;miRanda软件预测miR-29a的靶基因,western blot验证其表达结果。 结果：荧光定量RT-PCR检测结果表明,与癌旁组织（2.17±0.89）比较,miR-29a在乳腺癌组织（5.65±1.45）中的表达水平上调（t=3.94,P＜0.01）;MTT实验和Transwell实验结果显示,与mimics阴性对照组［（106.36±5.15)%,(216.70±7.20）个］相比,miR-29a mimics组［（133.32±6.31）%,(294.30±8.60)个］乳腺癌细胞的增殖和迁移能力增加（t分别为3.36和6.85,P均＜0.01）;与inhibitor阴性对照组［（105.35±3.42）%,(240.30±9.50）个］相比,miR-29a inhibitor组［（66.63±3.82）%,(156.30±7.80）个］乳腺癌细胞的增殖和迁移能力降低（t分别为8.18和6.81,P均＜0.01）。miRanda软件预测人第10号染色体缺失的同源性磷酸酶-张力蛋白基因（PTEN）可能为miR-29a的靶基因。western blot结果显示,与mimics阴性对照组（0.55±0.01）相比,乳腺癌细胞转染miR-29a mimic后,其PTEN蛋白质的表达水平（0.22±0.01）下调（t=14.29,P＜0.01）;与inhibitor阴性对照组（0.49±0.02）相比,转染miR-29a inhibitor后,PTEN蛋白质的表达水平（1.25±0.02）上调（t=19.39,P＜0.01）。 结论：miR-29a在乳腺癌组织中的表达水平增高,并可能通过下调PTEN蛋白的表达促进乳腺癌细胞的增殖和转移。     

miR-27a基因多态性与乳腺癌易感性的Meta分析

目的：系统评价不同人群中微小RNAmiR‐27ars895819多态性与乳腺癌易感性的关系。方法计算机检索PubMed、EMBASE、CochraneLibrary、中国生物医学文献数据库（CBM）、中文科技期刊全文数据库（VIP）、中国期刊全文数据库（CNKI）及万方数据库,收集国内外发表的关于miR‐27a基因多态性与乳腺癌易感性的病例对照研究。检索时间均从建库至2014年6月,按纳入和排除标准筛选文献并评价纳入研究质量后,应用RevMan5．2软件进行Meta分析。结果该研究共纳入5篇文献,其中2篇在欧洲人群中进行研究,3篇在亚洲人群中进行研究。Meta分析结果显示：在总体人群与亚洲人群中miR‐27ars895819多态性与乳腺癌易感性无关,而在欧洲人群中有3个模型分析表明miR‐27ars895819多态性与乳腺癌易感性有关,分别为等位基因模型Gvs．A（OR=0．89,95CI％：0．82～0．97,P=0．008）;共显性模型AGvs．AA（OR=0．83,95％CI：0．74～0．94,P=0．002）;显性模型GG＋AGvs．AA（OR=0．83,95％CI：0．75～0．91,P=0．002）。结论在总体人群及亚洲人群中miR‐27rs895819多态性与乳腺癌易感性无关。然而在欧洲人群中miR‐27rs895819多态性与乳腺癌易感性有关。     

miR-544a Stimulates endometrial carcinoma growth via targeted inhibition of reversion-inducing cysteine-rich protein with Kazal motifs

Endometrial carcinoma (EC) is a female-specific malignant tumor. Although current treatments can achieve good outcomes and improve patient survival, there remains a high incidence of treatment-induced infertility, a serious side effect that is unacceptable to those of childbearing age. Studies have demonstrated that micro ribonucleic acids (microRNAs or miRNAs) such as miR-544a regulate tumor-related gene expression. However, whether miR-544a is involved in the progression of EC is unknown. This study aimed to investigate the biological functions and underlying mechanisms of miR-544a in EC in vivo and in vitro. Quantitative real-time polymerase chain reaction (qRT-PCR) revealed miR-544a overexpression in EC tissue and cell lines, which was associated with a decreased in overall survival as revealed by Kaplan–Meier analysis. Functionally, the miR-544a inhibitor restricted the proliferation [detected by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl-2-H-tetrazolium bromide (MTT) assay], invasion, and migration (detected by transwell assay) of human endometrial adenocarcinoma cells (HEC-1B and Ishikawa) and facilitated cell apoptosis (detected by flow cytometry assay). Western blotting analysis revealed that the miR-544a inhibitor decreased the expressions of matrix metalloproteinase (MMP)-2 and MMP-9 and elevated the levels of cleaved caspase3 and cleaved poly (ADP-ribose) polymerase. Furthermore, animal experiments indicated that the miR-544a antagonist (antagomir-544a) suppressed tumor growth significantly in a mouse xenograft model. The mechanistic, qRT-PCR, and immunohistochemical indications were that a reversion-inducing cysteine-rich protein with Kazal motifs (RECK) and miR-544a had inverse expression changes in EC. Bioinformatics analysis revealed RECK as a potential target for miR-544a, and this was verified by the dual-luciferase reporter assay. Subsequently, in vitro experiments, including transwell assay, MTT assay, flow cytometry assay, and Western blotting analysis, demonstrated that RECK exerted antitumor effects on EC, which were negatively regulated by miR-544a. Taken together, our study findings suggested miR-544a as a valuable target in EC therapy.     

PD-L1和miR-34a在三阴乳腺癌中的表达及相关性研究

目的探讨PD-L1和miR-34a在三阴乳腺癌(TNBC)中的表达、相关性及临床意义。方法选取三阴乳腺癌组织和癌旁组织标本各40例,RT-PCR检测miR-34a的相对表达量,免疫组化法检测PD-L1的表达,并进行统计学分析。结果PD-L1在TNBC细胞中有13例阳性(32.5%),表达阳性率高于癌旁组织(0),差异显著(P<0.05);miR-34a在TNBC细胞中的表达量是癌旁组织中的0.58倍(0.546±0.083 vs 0.940±0.097),差异显著(P<0.05);并且,miR-34a和PD-L1的表达呈负相关性(r=-0.403,P<0.05)。结论PD-L1在TNBC细胞中的表达阳性率高,miR-34a低,且miR-34a与PD-L1的表达呈负相关性,提示在TNBC中,miR-34a可能参与调控PD-L1的免疫应答反应。     

miR-1301 promotes prostate cancer proliferation through directly targeting PPP2R2C

Prostate cancer is the leading cause of cancer deaths among men in the worldwide, it’s important to find new prognostic factors and therapeutic targets. microRNAs play critical roles in the development and progression of prostate cancer. Here we revealed miR-1301 promoted prostate cancer progression. miR-1301 was upregulated in prostate cancer tissues and cells, overexpression of miR-1301 promoted anchorage-dependent and -independent growth using MTT analysis, colony formation analysis and soft agar growth analysis, whereas knockdown of miR-1301 suppressed anchorage-dependent and -independent growth. We also found overexpression of miR-1301 inhibited p27 expression and promoted Cyclin D1 expression, whereas knockdown of miR-1301 reduced this effect, suggesting miR-1301 promoted the G1/S transition. These results suggested miR-1301 promoted cell proliferation of prostate cancer. microRNAs can inhibit target mRNA translation or/and induce mRNA degradation, we found tumor suppresser PPP2R2C was the target of miR-1301, simultaneous downregualtion of PPP2R2C and miR-1301 promoted anchorage-dependent and -independent growth. These findings suggested miR-1301 promoted prostate cancer proliferation by inhibiting PPP2R2C, and might a therapeutic target for prostate cancer.     

miR-34a调控下游基因FUT8表达介导乳腺癌多药耐药的实验验证

目的探究miR-34a及FUT8的异常表达对乳腺癌多药耐药性的调控机制,明确乳腺癌耐药的诊疗靶点。方法采用Real-time PCR及western blot技术检测MCF-7和MCF-7/ADR中miR-34a和FUT8的表达水平; Real-time PCR技术检测乳腺癌细胞系中miR-34a的转染效率;通过生物信息学方法预测并采用双荧光素酶报告实验验证FUT8与miR-34a之间的靶向关系;特异性调控miR-34a的表达后,分别通过Real-time PCR、western blot以及免疫荧光染色检测转染细胞系中FUT8基因及蛋白质水平变化;采用CCK8实验、免疫荧光实验检测其对MCF-7和MCF-7/ADR细胞增殖、耐药性的调控作用。结果 miR-34a在MCF-7中的表达水平明显高于MCF-7/ADR(P=0.002 6);FUT8基因在MCF-7/ADR中的表达水平明显高于MCF-7(P=0.001 6);FUT8是miR-34a调控乳腺癌耐药性的靶点(P=0.001 9);特异性上调MCF-7/ADR细胞中miR-34a的水平可明显抑制FUT8的表达,并抑制该细胞的多药耐药性及增殖性;而下调MCF-7细胞中miR-34a表达后,FUT8的表达水平明显升高,同时增强了细胞多药耐药及增殖能力。结论 miR-34a通过调控下游基因FUT8的表达介导乳腺癌多药耐药。     

miR-34a与人乳腺癌细胞系MCF-7多柔比星耐药的关系

目的探讨miR-34a对人乳腺癌细胞系MCF-7多柔比星(Adr)耐药性的影响及其潜在分子机制。方法用低浓度逐步加量诱导法,建立MCF-7的Adr耐药细胞系(MCF-7/Adr),用miRNA芯片结合RT-qPCR筛选并验证miRNA在MCF-7/Adr及MCF-7细胞中的表达差异;用生物信息学软件对miR-34a进行基因靶向预测;通过miR-34a模拟物(mimic)和抑制物(inhibitor)转染实验结合MTT、实时荧光定量PCR(RT-qPCR)观察miR-34a的表达变化对细胞耐药性的影响,western blot检测转染后Notch1蛋白表达水平。结果成功构建MCF-7/Adr耐药亚系;与MCF-7细胞相比,MCF-7/Adr中有156个差异表达miRNA;其中miR-34a的表达水平明显降低(t=11.597,P=0.000)。与阴性对照组相比,转染miR-34a mimic后MCF-7/Adr细胞miR-34a表达水平增高(t=8.013,P=0.001),且增加了对Adr的敏感性(t=18.160,P=0.000);转染miR-34a inhibitor后MCF-7细胞miR-34a的表达水平降低(t=9.979,P=0.000),且降低了对Adr的敏感性(t=4.130,P=0.009)。靶基因预测结果显示,Notch1为miR-34a的特异性靶基因。western blot结果表明,与空白对照组和阴性对照组相比,MCF-7/Adr细胞转染miR-34a mimic后,Notch1蛋白的表达水平下降(F=64.949,P=0.000)。MCF-7细胞转染miR-34a inhibitor后,Notch1蛋白的表达水平升高(F=10.938,P=0.010)。结论 MCF-7/Adr及MCF-7细胞miRNA表达谱存在差异;miR-34a参与调节细胞对Adr的耐药过程,Notch1基因可能是调控靶基因之一。     

Let-7a mimic attenuates CCL18 induced breast cancer cell metastasis through Lin 28 pathway

BackgroundMicroRNAs are believed to influence breast cancer cell tumorgenicity by interacting with the production of tumor associated macrophages. At this stage, this hypothesis lacks sufficient empirical evidence. Our study is an investigation of the effects of let-7a on the function of human breast cancer cell lines that had undergone chemokine ligand 18 (CCL18) stimulation.MethodsTwo breast cancer cell lines MDA-MB-231 and MCF-7 were transfected with let-7a mimics with or without CCL18 simulation. The expression level of let-7a was evaluated with qRT-PCR. Our study examined cell proliferation, migration and cell cycles following let-7a treatment. The predicted target of let-7a was identified and confirmed in vitro by a dual luciferase reporter system. The associations between let-7a, CCL18 and target gene expression were evaluated using RT-PCR and the Western blotting method.ResultsThe downregulated expression level of let-7a was observed in both breast cancer cell lines. When compared to the control and CCL18 stimulation groups, cell proliferation and migration in MDA-MB-231 and MCF-7 cells were significantly inhibited by let-7a. Furthermore, the cell cycle was dramatically blocked at the G2/M phase. The luciferase reporter identified Lin28 as the direct binding target of let-7a in both breast cancer cell lines.ConclusionUpregulation of let-7a carries the potential to reverse CCL18 induced cell proliferation and migration alteration in breast cancer cells by regulating Lin28 expression. Our results provided evidence which suggests the use of let-7a as a therapeutic agent in the treatment of breast cancer.     

miR-21和miR-93表达水平与三阴性乳腺癌发生发展的相关性研究

目的探讨miR-21和miR-93的表达水平与三阴性乳腺癌(TNBC)发生发展的相关性。方法选择行乳腺癌根治术的女性患者86例,收集癌组织与正常乳腺组织,采用逆转录聚合酶链反应(RT-PCR)检测miR-21和miR-93的相对表达量,并分析其与临床病理特征的相关性。结果 miR-21和miR-93在TNBC组织中相对表达量高于癌旁组织;TNBC组织中,miR21高表达54例,占62.79%,miR-93高表达50例,占58.14%;miR-21和miR-93在肿瘤直径5 cm、临床分期为Ⅲ~Ⅳ期以及淋巴结转移的患者中高表达率显著增高(P0.01);miR-21和miR-93的表达水平与患者年龄、月经状态、病理分级无明显相关性。癌组织中miR-21与miR-93表达水平呈显著正相关(r=0.761,P0.01)。结论 miR-21与miR-93在TNBC中表达异常增高,且与肿瘤直径、临床分期、淋巴结转移有关,可作为TNBC诊断及判断预后的辅助指标。     

miR-1184 regulates the proliferation and apoptosis of colon cancer cells via targeting CSNK2A1

MicroRNA (miRNA) exerts an important part in colon cancer cell proliferation and apoptosis. Meanwhile, the dysregulation of some miRNAs is detected in colon cancer cells. However, it remains unclear about the underlying mechanism of their effects on tumor pathogenesis. The current work aimed to examine the miR-1184 effect on colon cancer cells. The differentially expressed miRNAs (DEMs), including miR-9-3p, miR-1184, miR-492, miR-92a-1-5p and miR-20a-3p, were obtained from the GSE115108 and GSE132619 data sets using the ‘GEO2R’ online tool. Based on the findings, miR-1184 was significantly down-regulated within colon cancer cells and tissues. Moreover, the experimental results of CCK8, flow cytometry, colony formation and Western blotting assays showed that, miR-1184 over-expression suppressed colon cancer cell proliferation through inhibiting Ki67 expression and promoted their apoptosis through up-regulating cleaved caspase-3 and down-regulating Bcl-2 expression. By contrast, miR-1184 inhibition exerted the opposite effects. A total of 110 target genes of miR-1184 were predicted using the TargetScan and miRTarBase databases, which were then used to construct the protein-protein interaction (PPI) network based on the DAVID and STRING websites and to perform GO and KEGG pathway enrichment analyses. The MCODE plug-in of cytoscape was utilized to verify that CSNK2A1 was the target gene and key gene in significant modules. MiR-1184 directly targets CSNK2A1 via using RNA immunoprecipitation assay and luciferase reporter gene assay. According to the results, CSNK2A1 over-expression reversed the functions of miR‐1184 over-expression in suppressing colon cancer cell proliferation and enhancing their apoptosis. In conclusion, over-expression of miR-1184 inhibits colon cancer cell proliferation but promotes their apoptosis through down-regulating CSNK2A1 expression.     

乳腺癌HER-2靶向治疗的现状与发展前景

目的人表皮生长因子受体2(HER2)是表皮生长因子受体(EGFR或Er Bb)家族中扮演中心角色的一个成员。本文对HER2靶向治疗的现状与发展方向作一综述。方法查阅Pubmed、中国知网、万方、维普等数据库与乳腺癌HER2靶向治疗方面相关的文章,共纳入52篇文献。结果 HER2信号能够调节细胞增殖、存活和分化,与乳腺癌、胃癌、肺癌与卵巢癌等多种肿瘤的发生发展密切相关。目前使用的HER2靶向治疗有助于改善临床化疗药物疗效、提高患者无疾病生存期和5年存活率,但肿瘤的耐药性也越来越严重,已经引起人们的关注。结论 HER2靶向治疗对提高HER2阳性乳腺癌患者治疗效果起到重要作用,其耐药性需要进一步研究解决。