宫颈脱落细胞miR34a检测在HR-HPV阳性患者 分流中的作用分析

目的 探讨宫颈脱落细胞miR34a检测在HR-HPV阳性患者分流中的作用。方法 采用第二代杂交捕获技术（HC2）进行宫颈癌初筛,HPV阳性者行TCT及miRNA检查。TCT结果异常者（≥意义不明确的非典型鳞状细胞）阴道镜下活检行病理学检查。TCT阴性者行阴道镜检查,阴道镜异常者阴道镜下活检取材,比较不同级别宫颈病变中miR34a水平,探讨miR34a、Pap在宫颈病变中的诊断作用。结果 ①Pap对宫颈高度鳞状上皮内病变预测值的灵敏度为64.90%,特异度为80.70%,AUC为0.728（95%CI:0.657~0.779,P＜0.001）。②miRNA34a对宫颈高度鳞状上皮内病变预测值的灵敏度为77.00%,特异度为62.30%,AUC为0.824（95%CI:0.756~0.892,P＜0.001）,cut off值为0.623。③miRNA34a对宫颈高度鳞状上皮内病变预测值的灵敏度高于Pap检测,特异度低于Pap。④宫颈脱落细胞miRAN34a值随着宫颈病变加重逐渐降低。结论 检测宫颈脱落细胞miR34a水平可有效分流HR-HPV阳性患者,敏感性高于Pap检查。     

非小细胞肺癌组织miRNA-34b/c甲基化及预后相关性

目的探讨miRNA-34b/c在可手术的非小细胞肺癌组织中的甲基化及其与预后的相关性。方法回顾性分析中国医科大学附属盛京医院2009年12月-2015年12月期间行手术治疗的53例非小细胞肺癌患者的临床资料。采用定量RT-PCR检测上述病例肺癌标本中miRNA-34b/c的甲基化水平,应用Kaplan-Meier方法进行单因素生存曲线分析miRNA-34b/c甲基化水平与非小细胞肺癌生存的相关性,Log-rank法进行检验,采用Cox风险比例模型分析预后的独立影响因素。结果非小细胞肺癌miRNA-34b/c甲基化水平较高,生存分析显示miRNA-34b/c甲基化组生存期明显低于非甲基化组(P0.01),Cox风险比例模型分析显示miRNA-34b/c甲基化可作为非小细胞肺癌患者预后的独立影响因素。结论非小细胞肺癌miRNA-34b/c甲基化水平高,miRNA-34b/c甲基化提示非小细胞肺癌患者预后不良。     

miR-34a 通过Snail 诱导肺癌EMT 及促进其转移的分子机制

目的:探讨miR-34a 在肺癌组织中的表达情况以及miR-34a 在肺癌细胞侵袭和迁移过程中的作用及其机制。方法:qPCR 检测肺癌和正常肺组织中miR-34a 的表达情况;使用miR-34a-mimic 和miR-34a-inhibitor 过表达和沉默miR-34a,qPCR 检测沉默和过表达效果;Western blot 检测沉默和过表达miR-34a 后Snail 蛋白的表达情况;荧光素酶报告基因检测miR-34a 与Snail 的相互作用;Transwell 侵袭实验检测miR-34a 的表达对肺癌细胞侵袭能力的影响;划痕实验检测miR-34a 的表达对肺癌细胞迁移能力的影响;Western blot 检测E-Cadherin、Vimentin 和Twist 蛋白的表达情况。结果:与正常肺组织相比,肺癌组织中miR-34a 表达明显降低;且晚期、低分化和有淋巴结转移的肺癌组织miR-34a 表达明显较早期、高分化和无淋巴结转移的肺癌组织低;miR-34a-mimic miR-34a-inhibitor 可以有效抑制和过表达miR-34a 的表达;miR-34a 能与Snail 的3忆UTR 特异性结合;miR-34a 可以调控肺癌H1650 细胞的侵袭迁移能力;过表达miR-34a 上调E-Cadherin,同时下调Vimentin 和Twist 蛋白的表达,沉默miR-34a 则相反。结论:miR-34a 在肺癌中表达明显降低,且跟肺癌分期分级以及淋巴结转移与否密切相关,同时miR-34a 可以通过上皮间质转化调节肺癌细胞侵袭和迁移能力。     

miR-34a通过Snail诱导肺癌EMT及促进其转移的分子机制北大核心CSCD

目的:探讨miR-34a在肺癌组织中的表达情况以及miR-34a在肺癌细胞侵袭和迁移过程中的作用及其机制。方法:q PCR检测肺癌和正常肺组织中miR-34a的表达情况;使用miR-34a-mimic和miR-34a-inhibitor过表达和沉默miR-34a,q PCR检测沉默和过表达效果;Western blot检测沉默和过表达miR-34a后Snail蛋白的表达情况;荧光素酶报告基因检测miR-34a与Snail的相互作用;Transwell侵袭实验检测miR-34a的表达对肺癌细胞侵袭能力的影响;划痕实验检测miR-34a的表达对肺癌细胞迁移能力的影响;Western blot检测E-Cadherin、Vimentin和Twist蛋白的表达情况。结果:与正常肺组织相比,肺癌组织中miR-34a表达明显降低;且晚期、低分化和有淋巴结转移的肺癌组织miR-34a表达明显较早期、高分化和无淋巴结转移的肺癌组织低;miR-34a-mimic和miR-34a-inhibitor可以有效抑制和过表达miR-34a的表达;miR-34a能与Snail的3'UTR特异性结合;miR-34a可以调控肺癌H1650细胞的侵袭迁移能力;过表达miR-34a上调E-Cadherin,同时下调Vimentin和Twist蛋白的表达,沉默miR-34a则相反。结论:miR-34a在肺癌中表达明显降低,且跟肺癌分期分级以及淋巴结转移与否密切相关,同时miR-34a可以通过上皮间质转化调节肺癌细胞侵袭和迁移能力。     

miR-34a-sirt1-NRF2信号通路对阿霉素心脏损伤的机制研究

目的 探讨mi R-34a-sirt1-NRF2信号通路对阿霉素心脏损伤的影响。方法 构建阿霉素心脏损伤C57BL/6小鼠模型,构建模型前转染mi R-34a调控小鼠体内mi R-34a的水平。比较各组小鼠sirt1、NRF2蛋白表达水平,检测各组小鼠心脏血流动力学指标(+dp/dt_max、-dp/dt_max、LVEF、FS)水平。随后处死小鼠,检测外周血CD4⁺、CD8⁺、CD4⁺/CD8⁺水平以及血清CK-MB、LDH及心肌组织MDA、SOD水平。生物信息学预测mi R-34a的靶基因并用双荧光素酶报告基因进一步验证,检测心肌细胞（H9C2）mi R-34a过表达或沉默sirt1表达时凋亡蛋白（Bax、Bcl-2）及sirt1、NRF2蛋白的水平。结果 相较于正常对照小鼠,阿霉素干预的小鼠CD8⁺、血清CK-MB、LDH及心肌组织MDA水平显著上升,而CD4⁺、CD4⁺/CD8^...     

LncRNAs在肾癌中的作用机制及免疫治疗发展趋势

随着生物信息学技术的进步、高通量测序的应用,发现了大量的功能性的lncRNAs.它可以表现为抑癌基因、致癌基因甚至是同一LncRNAs在不同的环境及组织中表现出抑癌、致癌双重功能,并参与肿瘤的发生发展.研究表明,LncRNAs在肿瘤细胞的增殖、凋亡、侵袭、迁移以及肿瘤的转移及耐药等多种生物学进程中起着关键性的作用,因而有望成为肿瘤生物标记物以及药物治疗新靶点用于肿瘤诊断、治疗、预后预测.     

microRNA-155在免疫系统中的作用

microRNA( miRNAs,miRs)是一组近几年发现的、不编码蛋白质的单链小分子RNA.其在疾病发生发展过程中的作用是目前研究的热点.miRNA在免疫系统中的作用已经被广泛的研究,其不仅参与免疫系统的发生发展,而且在介导免疫系统抵抗微生物入侵机体方面起重要的作用.miR-155是免疫系统最主要的miRNA之一,其在活化的各种免疫细胞中表达普遍升高,是参与天然免疫应答和特异性免疫应答的最重要的miRNA.现就miR-155在免疫系统中的作用做一综述.     

microRNA-30a调控Foxo-1表达在子宫内膜异位症发病中的作用

目的 探讨microRNA-30a调控Foxo-1表达在子宫内膜异位症发病中的作用。方法 qRT-PCR检测microRNA-30a和Foxo-1 mRNA水平,将分离纯化和培养后的异位内膜基质细胞进行分组,分别为对照组、microRNA-30a组和microRNA-30a抑制剂组,CCK-8检测细胞增殖能力,Transwell法检测细胞迁移和侵袭,酶联免疫吸附实验（ELISA）检测细胞炎症反应,Western blot检测各组细胞Foxo-1、p53、p65和VEGF蛋白水平。结果 与对照组相比,在位内膜组和异位内膜组中的microRNA-30a和Foxo-1 mRNA水平显著降低,其中异位内膜组与对照组相比,microRNA-30a和Foxo-1 mRNA水平明显更低;microRNA-30a组细胞增殖、迁移和侵袭能力显著低于对照组,microRNA-30a抑制剂组细胞增殖、迁移和侵袭能力明显高于对照组和microRNA-30a组;microRNA-30a组中炎症因子TNF-a、IL-1β、IL-16和IL-18水平显著低于对照组,microRNA-30a抑制剂组中炎症因子TNF-...     

Upregulation of MicroRNA-34a Sensitizes Ovarian Cancer Cells to Resveratrol by Targeting Bcl-2

PurposeResveratrol (REV), a natural compound found in red wine, exhibits antitumor activity in various cancers, including ovarian cancer (OC). However, its potential anti-tumor mechanisms in OC are not well characterized. Here, we tried to elucidate the underlying mechanisms of REV in OC cells.Materials and MethodsThe anti-proliferative effects of REV against OC cells were measured using CCK-8 assay. Apoptosis was measured using an Annexin V-FITC/PI apoptosis detection kit. The anti-metastasis effects of REV were evaluated by invasion assay and wound healing assay. The miRNA profiles in REV-treated cells were determined by microarray assay.ResultsOur results showed that REV treatment suppresses the proliferation, induces the apoptosis, and inhibits the invasion and migration of OV-90 and SKOV-3 cells. miR-34a was selected for further study due to its tumor suppressive roles in various human cancers. We found miR-34a overexpression enhanced the inhibitory effects of REV on OC cells, whereas miR-34a inhibition had the opposite effect in OC cells. In addition, we verified that BCL2, an anti-apoptotic gene, was found directly targeted by miR-34a. We also found that REV reduced the expression of Bcl-2 in OC cells. Further investigations revealed that overexpression of Bcl-2 significantly abolished the anti-tumor effects of REV on OC cells.ConclusionOverall, these results demonstrated that REV exerts anti-cancer effects on OC cells through an miR-34a/Bcl-2 axis, highlighting the therapeutic potential of REV for treatment of OC.     

miR-34a在结直肠癌中的表达及其临床意义

目的 探讨miR-34a在结直肠癌中的表达及其与临床病理特征的关系.方法 应用实时荧光定量PCR法对43例结直肠癌及其癌旁正常组织中miR-34a进行定量检测.结果 结直肠癌组织中miR-34a表达水平明显高于癌旁正常组织,差异有统计学意义(P＞0.05);miR-34a在结直肠癌组织中的表达与患者性别、年龄、肿瘤部位、肿块大小、淋巴结转移均无相关性(P＞0.05),而与浸润深度、TNM分期密切相关(P＜0.05).结论 miR-34a在结直肠癌的发生、发展过程可能发挥重要作用,检测miR-34a有望成为结直肠癌新的肿瘤标记物或预后因子.     

MicroRNA-34a negatively regulates anesthesia-induced hippocampal apoptosis and memory impairment through FGFR1

Background: Mounting evidence has shown the toxic effects of anesthesia to neonatal hippocampus. We used an in vivo mouse model to explore the role of microRNA 34a (miR-34a) in regulating anesthesia-induced hippocampal neurotoxicity. Methods: One-month old C57/BL6 mice received daily intraperitoneal injection of anesthesia (ketamine, 50 mg/kg) for 7 days. One day after, apoptosis was evaluated by TUNEL staining in hippocampal CA1 region, and expression level of miR-34a assessed by real-time quantitative PCR (qPCR). Hippocampal miR-34a was then down-regulated through lentivirus mediated cortical injection prior to anesthesia. The effects of inhibiting hippocampal miR-34a on anesthesia-induced hippocampal apoptosis and memory impairment were further investigated by TUNEL staining and Morris water maze (MWM) test. The predicted molecular target of miR-34a, fibroblast growth factor receptor 1 (FGFR1) was down-regulated in hippocampus through siRNA-mediated cortical injection and its effect on hippocampal apoptosis was also examined. Results: Anesthesia caused severe apoptosis among hippocampal CA1 neurons and upregulated hippocampal miR-34a. On the other hand, lentivirual inhibition of miR-34a protected anesthesia-induced hippocampal apoptosis and memory impairment. Luciferase essay demonstrated FGFR1 was directly regulated by miR-34a in hippocampus. siRNA-induced FGFR1 downregulation further exaggerated anesthesia-induced apoptosis in hippocampus. Conclusions: Overall, we showed that miR-34a negatively modulated anesthesia-induced hippocampal neurotoxicity.     

白藜芦醇与奥沙利铂联合应用抑制结直肠癌细胞增殖的机制

目的:探讨联合应用白藜芦醇与奥沙利铂抑制结直肠癌细胞系增殖的作用机制。方法:单独或联合应用奥沙利铂和白藜芦醇处理人结肠癌细胞系HCT-116和HT-29,CCK-8实验检测肿瘤细胞活力,流式细胞术检测细胞周期;应用慢病毒介导使HCT-116细胞过表达miR-34c,再用奥沙利铂处理,检测肿瘤细胞的活力和凋亡;建立结直肠癌皮下荷瘤裸鼠模型,分别给予二甲基亚砜、奥沙利铂、白藜芦醇及白藜芦醇+奥沙利铂,制作肿瘤组织石蜡切片进行Ki67染色并计数。结果:与单独应用奥沙利铂或白藜芦醇相比,联合应用奥沙利铂和白藜芦醇更显著抑制HCT-116细胞的活力,细胞阻滞在G1期。荷瘤裸鼠实验同样显示奥沙利铂和白藜芦醇联合应用较单独应用时Ki67阳性率减少。结论:联合应用奥沙利铂和白藜芦醇具有协同抑制结直肠癌细胞增殖的作用,其可能机制与白藜芦醇促进结直肠癌细胞抑癌基因miR--34c表达有关。     

MiR-34a inhibits oral cancer progression partially by repression of interleukin-6-receptor

Previous reports revealed that a significant decrease of miR-34a in oral cancer. But the role of miR-34a in oral cancer needs further research. In the present study, we will investigate the effect of miR-34a on oral cancer cell phenotypes. First, it was verified that miR-34a expression was lower in oral cancer tissues compared with their normal controls, so did the oral cancer cells. Next, it was showed that miR-34a overexpression in oral cancer cells could inhibit cell proliferation, G1 phase arrest, metastasis and epithelial mesenchymal transition. It was predicted that interleukin-6-receptor (IL6R) was a potential target gene of miR-34a by bioinformatics analysis and identified by luciferase assay. It was further showed that miR-34a inhibited oral cancer progression via IL6R. Collectively, our findings suggested that miR-34a may function as a tumor suppressor in oral cancer by targeting IL6R.     

MicroRNA-34a靶向SIRT1参与阿霉素诱导的心肌细胞凋亡

目的:研究微小RNA-34a(microRNA-34a,miR-34a)在阿霉素诱导的心肌细胞凋亡中的作用及其作用靶基因。方法:建立阿霉素(doxorubicin,Dox)诱导的大鼠H9c2心肌细胞凋亡模型;TUNEL染色观察H9c2细胞凋亡;双萤光素酶报告实验检测miR-34a与潜在靶基因沉默信息调节因子1(silent information regulator 1,SIRT1)3'端非翻译区(3'-untranslated region,3'UTR)的结合作用;实时荧光定量PCR检测miR-34a和SIRT1 mRNA表达水平,Western blot检测SIRT1和凋亡相关蛋白表达水平。结果:阿霉素处理H9c2细胞之后,细胞发生凋亡,miR-34a的表达显著增强;双萤光素酶报告实验提示miR-34a与SIRT1 3'UTR可相互作用,并证实miR-34a可在转录后水平抑制SIRT1的表达,SIRT1蛋白水平在阿霉素处理的心肌细胞中显著下调;过表达miR-34a及沉默SIRT1均能一致性抑制Bcl-2表达,促进Bax和p66shc的表达,而过表达SIRT1能有效抑制阿霉素诱导的H9c2细胞凋亡。结论:SIRT1是miR-34a的靶基因,并介导了miR-34a在阿霉素诱导的心肌细胞凋亡中的作用。     

MicroRNA-34a inhibits tumor invasion and metastasis in gastric cancer by targeting Tgif2

To study how miR-34a acts as a tumor suppressor in inhibiting the invasion and metastasis of the gastric cancer cells. First, real-time polymerase chain reaction (PCR) and western blot analysis were used to analyze the expression of miR-34a and Tgif2 in gastric cancer tissues and the adjacent normal tissues. Next, gastric cancer cells were transfected with miR-34a mimic and Tgif2 siRNA, respectively. After transfection, real-time PCR and western blot analysis were used to detect the relative Tgif2 expression level. Cell proliferation was monitored by the colorimetric water-soluble tetrazolium salt and apoptosis analysis was performed with Annexin-V-FITC Apoptosis Detection Kit I. The expression of miR-34a in the adjacent non-tumor tissues was higher than that in gastric cancer tissues, but Tgif2 was opposite. In gastric cancer cells transfected with miR-34a mimic/Tgif siRNA, Tgif2 expression was remarkably down-regulated. Cells transfected with miR-34a mimic/Tgif2 siRNA grew more slowly than the control groups. The percentage of apoptotic cells in gastric cancer cells transfected with miR-34a mimic/Tgif siRNA was much higher compared to the controls. Therefore, we concluded that miR-34a could inhibit tumor invasion and metastasis in gastric cancer by targeting Tgif2 and may be a novel therapeutic candidate for gastric cancer.     

PABPC1 exerts carcinogenesis in gastric carcinoma by targeting miR-34c

As one of the common malignant tumors that threaten human health severely, gastric carcinoma is the second highest cause of cancer death and the fourth most common cancer globally. However, the mechanism underling gastric cancer is still not fully understood. PABPC1 plays an important role in translation, control the rate of mRNA deadenylation and participates in mRNA decay, which is involved in carcinogenesis. Here in present study, we reported that PABPC1 is an oncogenic protein in gastric carcinoma. The results showed that PABPC1 is upregulated in gastric carcinoma tissues, and high PABPC1 expression predicts poor survival. PABPC1 regulates proliferation and transformation of gastric cancer cells in vitro and in vivo. PABPC1 knockdown induces apoptosis by upregulating pro-apoptotic proteins and downregulating anti-apoptotic proteins. In addition, miR-34c is a target of PABPC1, and miR-34c is critically essential for the function of PABPC1. In summary, PABPC1 exerts carcinogenesis and promotes growth and survival of gastric cancer cells by regulating miR-34c.     

miR-34a调控的M2型巨噬细胞极化对食管癌细胞迁移和凋亡的影响

目的探究miR-34a在调控M2型巨噬细胞极化中的作用及其对食管癌细胞迁移和凋亡的影响。方法实时荧光定量PCR检测不同分化状态下的巨噬细胞miR-34a的表达水平;在M2型巨噬细胞中转染对照microRNA（miR-NC）和miR-34a mimics,使用荧光定量PCR检测CD163、CD206、CCL18和CCL22 m RNA的表达水平,使用酶联免疫吸附实验检测培养基中IL-10和TGF-β的表达水平;细胞划痕实验检测EC109细胞迁移能力;流式细胞术检测5-氟尿嘧啶（5-FU）诱导的EC109细胞凋亡水平;MTT法检测5-FU的半数抑制计量(IC₅₀)。结果与THP-1和M0型巨噬细胞相比,M2型巨噬细胞极化过程中miR-34a表达水平显著下降;与对照组细胞相比,转染miR-34a mimics的M2巨噬细胞CD163、CD206、CCL18和CCL22 mRNA表达水平显著下降,分泌到培养基中的IL-10和TGF-β含量显著下降。与对照组相比,使用过表达miR-34a的M2巨噬细胞条件培养基培养的食管癌EC109细胞迁移水平显著下降,5-FU诱导的细胞凋...     

Identification of microRNA expression signature for the diagnosis and prognosis of cervical squamous cell carcinoma

Cervical cancer is the fourth leading cause of cancer death among women globally. The prognosis of cervical cancer patients differs considerably, and clinical outcomes are difficult to predict. Given the significant roles of miRNAs in human cancers, identification of novel and reliable miRNA biomarkers is important for targeted cervical cancer therapy. In the present study, we aimed to reveal biological significance of miR-200a, miR-423, miR-34a, miR-193a, and miR-455 for the prognosis and diagnosis of cervical cancer and their association with the clinical outcomes of patients. Distinct expression profiles of miRNAs in formalin-fixed paraffin-embedded tissue samples of patients and healthy controls were evaluated using qRT-PCR. We identified miR-200a, miR-455, and miR-34a were significantly downregulated in cervical squamous cell carcinoma tissues compared to normal cervix tissue from healthy controls. Both miR-455 and miR-34a confer a promising diagnostic factor for the cervical cancer while miR-200a showed no significance in ROC analysis. Notably, low expression of miR-34a was markedly associated with the poor overall survival of cervical cancer patients as revealed by Kaplan-Meier survival analysis. Also, univariate and multivariate analysis indicated miR-34a expression as an independent prognostic factor. Consequently, our results underline the importance of distinct expression miRNAs in cervical squamous cell carcinoma.