胃癌患者血清miR-135与miR-601水平变化及临床意义

目的探讨胃癌患者血清miR-135与miR-601水平变化及临床意义。方法选择2019-2020年唐山市第三医院收治的90例胃癌患者作为观察组,另选择同期84例体检健康者作为对照组,检测并比较2组血清miR-135、miR-601、糖类抗原72-4(CA72-4)、糖类抗原19-9(CA19-9)水平,采用受试者工作特征(ROC)曲线评估血清miR-135、miR-601、CA72-4、CA19-9对胃癌的早期诊断及预后评估效能。结果观察组血清miR-135、miR-601、CA72-4、CA19-9水平高于对照组(P<0.05),进展期胃癌患者血清miR-135、miR-601、CA72-4、CA19-9水平高于早期胃癌患者(P<0.05)。血清miR-135、miR-601水平与胃癌临床分期及肿瘤分化程度有关(P<0.05),与年龄、性别、肿瘤最大径、组织学类型以及肿瘤位置无关(P>0.05)。ROC曲线结果显示,miR-135、miR-601、CA72-4、CA19-9联合检测诊断早期胃癌的曲线下面积(AUC)为0.961,灵敏度为0.872,特异度为0.835,优于单独检测的AUC(0.952、0.943、0.557、0.677)、灵敏度(0.820、0.845、0.799、0.812)和特异度(0.796、0.803、0.748、0.790);miR-135、miR-601、CA72-4、CA19-9联合检测评估胃癌预后的AUC为0.953,灵敏度为0.890,特异度为0.843,优于单独检测的AUC(0.948、0.675、0.566、0.625)、灵敏度(0.834、0.825、0.785、0.811)和特异度(0.810、0.832、0.767、0.807)。结论血清miR-135和miR-601水平在胃癌患者中明显升高,二者联合CA72-4、CA19-9水平检测对胃癌的早期诊断和预后评估具有较高价值,有望成为胃癌早期诊断和预后评估的新型血清标志物。     

TMPRSS4通过上皮间质转化促进乳腺癌细胞的增殖、侵袭和转移

目的通过检测Ⅱ型跨膜丝氨酸蛋白酶4（TMPRSS4）在乳腺癌细胞中的表达情况,分析其在乳腺癌细胞增殖、侵袭和转移过程中的作用及其与上皮间质转化（EMT）的关系。 方法采用实时荧光定量PCR（qRT-PCR）和Western blot法检测5种不同乳腺癌细胞系中TMPRSS4 mRNA和蛋白水平的表达情况。将过表达质粒转染至乳腺癌细胞中,采用qRT-PCR方法检测转染效率,并通过MTS和EdU细胞增殖实验、Transwell和Matrigel细胞迁移和侵袭实验,研究过表达TMPRSS4对乳腺癌细胞增殖、侵袭和迁移的作用。采用qRT-PCR和Western blot法检测过表达TMPRSS4后细胞EMT相关基因E-cadherin、Vimentin、Claudin-1、Slug、ZEB1的表达变化。 结果TMPRSS4在乳腺癌MDA-MB-468和MDA-MB-231细胞系中高表达。MTS实验显示TMPRSS4过表达组乳腺癌细胞MDA-MB-468和MDA-MB-231的增殖能力与阴性对照组相比明显增加,差异具有统计学意义（P=0.039和0.038）,EdU实验进一步证实过表达TMPRSS4促进乳腺癌细胞的增殖,MDA-MB-468细胞和MDA-MB-231细胞的增殖率与阴性对照组比较差异具有统计学意义（P=0.001和0.008）。Transwell实验显示,TMPRSS4过表达组MDA-MB-468细胞和MDA-MB-231细胞的迁移能力与阴性对照组比较明显提高,差异具有统计学意义（p均=0.001）。Matrigel实验显示,TMPRSS4过表达组MDA-MB-468细胞和MDA-MB-231细胞的侵袭浸润能力与阴性对照组比较明显提高,差异具有统计学意义（P=0.012和0.000）。与阴性对照组比较,过表达TMPRSS4后,EMT相关基因包括上皮性标记E-cadherin和Claudin-1的表达均显著下降,而间质标记Vimentin和Slug的表达均明显提高,差异具有统计学意义（P分别=0.024,0.003,0.002和0.012）。 结论TMPRSS4参与了乳腺癌EMT的发生过程,并通过EMT促进乳腺癌细胞的增殖、侵袭和转移。     

血清miR-135、miR-601、CA72-4及CA19-9的表达水平与胃癌诊断的相关性

目的 探究血清miR-135、miR-601、糖类抗原72-4(CA72-4)及糖类抗原19-9(CA19-9)的表达水平与胃癌(GC)诊断的相关性.方法 选取2015年1月至2020年10月本院106例GC患者作为观察组,健康人群45例作为对照组,检测并比较两组血清miR-135、miR-601、CA72-4及CA1...     

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...     

LncRNA PTPRG-AS1通过靶向miR-5590-3p调控乳腺癌细胞增殖、迁移和侵袭的研究

目的 探讨LncRNA PTPRG-AS1对乳腺癌细胞增殖、迁移和侵袭的影响及可能机制.方法 以正常乳腺上皮细胞MCF-10A为对照,qRT-PCR法检测乳腺癌细胞系(MCF-7、T47D和BT549)中LncRNA PTPRG-AS1和miR-5590-3p表达.分别转染si-LncRNA PTPRG-AS1、miR...     

斯钙素-1在乳腺癌中的表达及临床意义

目的：从mRNA和蛋白水平检测斯钙素-1（STC-1）在乳腺癌细胞系及乳腺癌患者外周血中的表达,并探讨其临床意义。方法2012年7月至2013年1月期间,应用RT-PCR检测3个乳腺癌细胞系、沈阳军区总医院收治的56例乳腺癌患者及10例健康女性外周血STC-1 mRNA的表达;应用酶联免疫吸附实验法检测STC-1蛋白的表达。结果在mRNA和蛋白水平,乳腺癌细胞系及患者外周血中STC-1表达均明显高于健康女性;且在不同TNM分期表达有显著差异（P<0.05）,Ⅳ期显著高于Ⅰ、Ⅱ、Ⅲ期（P<0.05）。STC-1 mRNA在小叶癌中的表达较导管癌及混合癌高（P=0.03）。STC-1 mRNA 和蛋白在CEA>5 ng/ml时表达较CEA≤5 ng/ml时明显增高（P<0.05）。结论 STC-1有望成为乳腺癌复发转移的标志物,其表达水平对于乳腺癌预后预测具有重要价值。     

miR-451a suppresses the development of breast cancer via targeted inhibition of CCND2

Extensive research has indicated that miRNAs are crucial for the occurrence and progression of cancers. miR-451a, involved in breast cancer (BC), is one of the miRNAs. This study focused on the mechanism by which miR-451a regulates BC. The levels of miR-451a in BC tissues and cell lines were examined using quantitative real-time polymerase chain reaction (qRT-PCR). Kaplan‒Meier analysis showed that this was intimately related to the patient's overall survival rate. Functional experiments revealed the negative effects of miR-451a on the abilities of BC cells to multiply (tested by Cell Counting Kit-8), migrate (tested by wound healing assay), and invade (tested by Transwell assay) and its positive effects on apoptosis (tested by flow cytometry). Western blotting indicated that the expression of tumor-related proteins was affected by miR-451a. Moreover, in vivo experiments suggested that tumor growth was clearly restrained by an miR-451a agonist in a xenograft tumor model. Bioinformatic analysis indicated that miR-451a directly targeted Cyclin D2 (CCND2), as demonstrated by the luciferase reporter assay. An opposite change in the level of CCND2 and miR-451a in BC was indicated by qRT-PCR, western blotting, and immunohistochemistry. Subsequently, functional experiments and western blotting analysis confirmed that CCND2 accelerated BC progression, which was regulated by miR-451a. Cumulatively, research on miR-451a may be valuable for BC treatment.     

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.     

miR-421靶向PDCD4促进前列腺癌DU145细胞增殖的实验研究

目的研究MicroRNA-421(miR-421)促进前列腺癌DU145细胞增殖的作用及潜在的分子机制。方法培养前列腺癌DU145细胞,分为对照组、miR-阴性对照(NC)组、miR-421组、miR-421+pcDNA组、miR-421+pcDNA-细胞程序性死亡基因4(PDCD4),miR-NC组转染miR-NC、miR-421组转染miR-421、miR-421+pcDNA组转染miR-421及pcDNA质粒、miR-421+pcDNA-PDCD4组转染miR-421及pcDNA-PDCD4质粒,MTS法测定细胞增殖活力,荧光定量PCR测定PDCD4的mRNA表达水平,western blot测定PDCD4的蛋白表达水平,双荧光素酶报告基因实验验证miR-421与PDCD4的靶向结合。结果与对照组及miR-NC组比较,miR-421组的增殖活力增加,PDCD4的表达水平及野生型PDCD4双荧光素酶报告基因的荧光活力均降低(P<0.05);与miR-421+pcDNA组比较,miR-421+pcDNAPDCD4组的增殖活力降低,PDCD4的表达水平增加(P<0.05)。结论 miR-421对前列腺癌DU145细胞的增殖具有促进作用,靶向抑制PDCD4是miR-421发挥这一作用的潜在分子机制。     

miR-195 inhibits tumor growth and angiogenesis through modulating IRS1 in breast cancer

Angiogenesis has been found as an attractive target for drug therapy as it is necessary for tumor growth. Accumulating evidences show that microRNAs (miRNAs), which are a group of highly conserved, single-stranded, short non-coding RNAs, play important roles through directly targeting angiogenic factors and protein kinases. The purpose of this study is to investigate the role of miR-195 in breast cancer development and angiogenesis through targeting IRS1. We show that miR-195 is inversely related with Insulin receptor substrate 1 (IRS1) in both breast cancer cells and breast cancer tissues. Induction of miR-195 could suppress IRS1 protein expression through binding to its 3′UTR regions either by transfection with miR-195 oligo or by infection with lentivirus encoding miR-195 gene. Moreover, re-expression of IRS1 reverses miR-195-mediated repression of tumor cell growth and miR-195 inhibits tumor angiogenesis through suppressing IRS1-VEGF axis. These data suggest that miR-195 mimics are potential therapeutic agents for breast cancer diagnose.     

Brachyury promotes tamoxifen resistance in breast cancer by targeting SIRT1

Tamoxifen is effective for treating estrogen receptor-alpha (ERα)-positive breast cancers. However, few molecular mediators of tamoxifen resistance have been elucidated. In the present study, we determine the underlying roles of Brachyury in tamoxifen resistance. Loss- and gain-of-function assay are utilized to confirm the oncogenic roles of Brachyury in breast cancer. Compared with the normal MCF10A cells, Brachyury is commonly overexpressed in breast cancer cell lines. Knockdown of Brachyury inhibits tamoxifen resistance, whereas overexpression of Brachyury enhances tamoxifen resistance as demonstrated increased cell viability and reduced cell apoptosis. Mechanistically, we demonstrate for the first time that Brachyury mediates tamoxifen resistance by regulating Sirtuin-1 (SIRT1). Collectively, our data, as a proof of principle, indicate that Brachyury is a candidate marker for predicting the clinical efficacy of tamoxifen and targeting SIRT1 could overcome resistance to tamoxifen in breast cancer cells.     

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。     

LATS2 as a poor prognostic marker regulates non-small cell lung cancer invasion by modulating MMPs expression

Large tumor suppressor 2 (LATS2) plays significant roles in tumorigenesis and cancer progression. This study was aimed to analyze the correlation between LATS2 expression and clinicopathologic features and its prognostic significance in non-small cell lung cancer (NSCLC). LATS2 expression was examined in 73 NSCLC clinical specimens and 22 normal lung tissues using immunohistochemistry. Low levels of LATS2 protein were inversely associated with the T classification (P = 0.001), N classification (P = 0.005) and clinical stage (P = 0.001) in NSCLC patients. Patients with lower LATS2 expression had a significantly shorter overall survival than patients with high LATS2 expression. Multivariate analysis suggested that low expression of LATS2 was an independent prognostic indicator (P = 0.002) for the survival of patients with NSCLC. Furthermore, overexpression of LATS2 resulted in mobility inhibition in NSCLC cell lines A549 and H1299, and reduced protein level of matrix metalloproteinase-2 (MMP-2) and MMP-9. On the contrary, LATS2 siRNA treatment enhanced cell mobility and increased MMP-2 and MMP-9 protein expression level. In conclusion, low expression of LATS2 is a potential unfavorable prognostic factor and promoted cell invasion and migration in NSCLC.     

MicroR-760 suppresses cancer stem cell subpopulation and breast cancer cell proliferation and metastasis: By down-regulating NANOG

Background and objectiveEmerging evidences suggest that cancer stem cells are responsible for tumor aggressive, metastasis and therapeutic resistance. To data, the mechanism underlying breast cancer stem cell (BCSC) population within tumor metastasis remains to be fully elucidated. The current study was to investigate the potential role of microRNA-760 (miR-760) and its associated target gene in population and metastasis of BCSC.MethodsCharacteristic BCSCs surface markers (CD44⁺/CD24^−/low) were determined by flow cytometry in breast cancer MCF-7 and BT-549 cells. Quantitative RT-PCR was used to evaluate miR-760 and NANOG mRNA expression. Expression of NANOG protein was determined using western blot. Cell proliferation was determined by MTT assay. The model of breast cancer cell xenograft was used to evaluate the effect of miR-760 on tumor growth.ResultsBT-549 cell has substantially more CD44⁺/CD24^−/low subpopulation than MCF-7 cell. Moreover, BT-549 cell expressed lower level of miR-760 and higher level of NANOG than MCF-7cell. By result from cellular miR-760 modulation, we found that miR-760 overexpression suppressed CD44⁺/CD24^−/low population as well as inhibited cell proliferation and migration of BT-549. On the contrary, knockdown of miR-760 promoted CD44⁺/CD24^−/low population and migration of MCF-7 cells. By luciferase reporter assay, miR-760 was proved to be functional associated with NANOG via regulating its expression. This functional interaction was showed to be involved in controlling proliferation and migration of MCF-7 and BT-549 cell.ConclusionThese data suggest that the target of miR-760/NANOG axis may represent a new therapeutic approach to suppress breast cancer stem cell subpopulation thereby prevent cancer metastasis.     

即刻早期反应基因-1在乳腺癌新辅助化疗后表达的变化及对预后的意义

目的探讨即刻早期反应基因-1(IEX-1)在乳腺癌新辅助化疗(NCT)后表达的变化及在预后判断中的意义。方法选取2012年1月至2017年1月首诊首治并接受NCT的110例乳腺癌女性患者作为研究对象进行回顾性研究,采用二步法免疫组化方法检测NCT前后乳腺癌组织中IEX-1表达的变化,采用Kaplan-Meier和Log-rank法分析其对无病生存期(DFS)及总生存期(OS)的影响。结果 NCT前乳腺癌组织中IEX-1蛋白均阴性;NCT后,IEX-1蛋白阳性表达45例(40. 9%),阴性65例。生存分析显示,IEX-1蛋白阳性表达组患者的DFS、OS均低于IEX-1蛋白阴性表达组(P均<0. 01)。多因素Cox比例风险回归模型分析发现,IEX-1蛋白阳性表达为乳腺癌预后的独立风险因素[HR=4. 921,95%CI=(1. 273~19. 018),P=0. 021]。结论乳腺癌NCT前后检测IEX-1有助于预后判断,IEX-1可能成为乳腺癌NCT疗效的预后指标。     

miR-663a inhibits hepatocellular carcinoma cell proliferation and invasion by targeting HMGA2

Hepatocellular carcinoma (HCC) is a highly aggressive solid malignancy throughout the world. Dysregulation of miRNAs play essential roles in HCC progression via aberrant regulation of cell proliferation, apoptosis, as well as metastasis. miR-663a is a poorly investigated miRNA. Whether miR-663a regulates HCC development remains unknown. The aim of the study was to explore the role of miR-663a in HCC development. To determine the expression level of miR-663a in HCC, we analyzed the data from GSE21362 and TCGA. The results showed that miR-663a was significantly down-regulated in HCC tissue compared with adjacent non-tumor tissue. Gain of function and loss of function assays revealed that miR-663a distinctly inhibited cell proliferation, migration and invasion. Mechanistic investigations demonstrated that miR-663a modulated cell functions through targeting and suppressing high mobility group A2 (HMGA2). In addition, overexpression of HMGA2 remarkably attenuated the tumor repressive effect of miR-663a. Taken together, miR-663a inhibits HCC cell proliferation and motility by targeting HMGA2.     

Evaluation of serum laminin as a tumor marker in breast cancer

Laminin is a noncollagenous constituent of the extracellular matrix (basement membrane). Increased serum concentrations were recorded in patients with a variety of cancers. The clinical usefulness of serum laminin as a marker for breast cancer was investigated in 60 female patients with malignant breast tumors (30 metastatic, 30 non-metastatic). Subjectively healthy age-matched women (n = 30) served as a control group. Laminin was significantly higher in breast cancer patients than in normal controls. Serum laminin levels were also significantly higher in patients with metastasis than in those without metastasis. A positive correlation was observed between serum laminin and the breast cancer-associated antigen CA 15-3 in the tumor patients. The sensitivity and specificity values of laminin for cancer detection at the optimum decision level [mean + 2 SD (1.4 U/ml)] were 75% and 97% respectively, with a 98% positive predictive value, 66% negative predictive value, and 82% diagnostic efficiency. For the detection of metastasis, serum laminin exhibited 77% sensitivity and 100% specificity [best decision level: mean + 2 SD (1.9 U/ml)], with a 100% positive predictive value, 81% negative predictive value, and 88% diagnostic efficiency. The latter specificity and positive predictive value were superior to those obtained with serum CA 15-3. These results suggest that serum determination of laminin could be a useful diagnostic tool in breast cancer and a valuable parameter in the prediction of metastasis.