Profilin 1, negatively regulated by microRNA-19a-3p,serves as a tumor suppressor in human hepatocellular carcinoma

Profilin 1 (PFN1) is a critical actin-regulatory protein; however, its functional role in hepatocellular carcinoma (HCC) progression remains to be further elucidated. In the present study, we observed that the expression levels of PFN1 were significantly decreased in HCC tissues and cell lines. Low PFN1 expression was significantly correlated with aggressive clinicopathological characteristics and poor prognosis of HCC patients. Further in vitro experiments demonstrated that overexpression of PFN1 remarkably inhibited the proliferation, migration, invasion and EMT of HCC cells. Moreover, we also found that PFN1 was a direct target gene of miR-19a-3p, and in HCC tissues, and there was a significantly inverse correlation between PFN1 mRNA and miR-19a-3p expression. Collectively, our results showed that PFN1 functions as a tumor suppressor in HCC, and might serve as a diagnostic and therapeutic target for HCC patients.     

胰腺导管腺癌中microRNA-21与TGF-β1信号通路相关

目的：对胰腺导管腺癌样本进行基因分析,筛选与胰腺导管腺癌相关的microRNA(miRNA),并初步分析目标miRNA与胰腺癌转化生长因子-β1(TGF-β1)信号通路的相关性。方法：收集在海南省中医院住院的胰腺导管癌患者术前外周静脉血血清标本19份,健康体检人群的外周静脉血血清标本21份作为非胰腺导管癌对照组。利用GCBI(Gene-Cloud Biotechnology Information)数据平台筛选与胰腺导管腺癌样本有关的基因并对其进行生物信息学分析,将筛选出的基因作为研究对象,用real-time PCR和蛋白质印迹法验证该基因在胰腺导管腺癌中的表达。通过改变胰腺导管腺癌细胞中该基因的表达水平观察其与TGF-β1之间的关系。结果：通过聚类分析和基因功能富集分析筛选出miRNA-21为胰腺导管癌相关基因。MiRNA-21在胰腺导管腺癌患者体内高表达。在miRNA-21过表达的PANC-1细胞中,TGF-β1表达受到抑制;但当miRNA-21的表达受到抑制时,TGF-β1的表达明显上升。结论：MiRNA-21在胰腺导管腺癌患者体内高表达,可调控TGF-β1的表达,从而参与胰腺导管腺癌的发生发展。     

MiR-30a-5p通过泛素水解酶22抑制人宫颈癌细胞上皮-间质转化功能

【目的】研究microRNA-30a-5p（miR-30a-5p）对人宫颈癌Hela细胞上皮-间质转化功能的影响及其相关机制。【方法】宫颈癌Hela细胞株分别转染目的mir的模拟物和阴性对照模拟物,分别以30a-5p组、NC组命名并标记细胞。同时,以未经过处理的Hela细胞作为对照（Control组）。分别用逆转录-聚合酶链反应法检测各组宫颈癌细胞的miR-30a-5p含量。Transwell实验检测3组细胞迁移能力和侵袭能力。Western-blot法检测3组细胞神经-钙粘素（N-cadherin）、α-连环蛋白（α-Catenin）和泛素水解酶22（USP22）表达水平。运用生物信息学方法预测miR-30a-5p的靶基因。采用Western blot法检测USP22过表达对miR-30a-5p抑制EMT的拮抗作用。双荧光素酶实验检测miR-30a-5p与USP22的关系。建立皮下移植瘤模型观察miR-30a-5p的体内作用。【结果】30a-5p组宫颈癌细胞miR-30a-5p的表达水平明显上调,表达水平为Control组的853.82（862.26~843.11）倍（P＜0.01）。30a-5p组侵袭细胞数量8.17（8.32~8.03）明显低于Control组（P＜0.01）。30a-5p组细胞N-cadherin蛋白的细胞内含量明显下降,α-Catenin蛋白的细胞内含量明显上升,USP22蛋白表达量明显降低。合并USP22过表达处理的30a-5p组宫颈癌细胞中N-cadherin蛋白表达量明显升高,α-Catenin蛋白表达量明显降低。双荧光素酶检验结果显示USP22为miR-30a-5p的下游靶基因（P＜0.01）。30a-5p组皮下移植瘤明显小于Control组（P＜0.01）。与Control组肿瘤组织相比,30a-5p组肿瘤组织miR-30a-5p的相对含量升高,USP22蛋白含量降低,N-cadherin蛋白的含量降低,α-Catenin蛋白含量升高。【结论】miR-30a-5p在宫颈癌Hela细胞中,可能通过靶向识别下游靶基因USP22,进而抑制其翻译。最终实现对宫颈癌细胞EMT过程的抑制。     

Dysregulation of microRNA-106a-5p-RUNX1 axis associates with clinical progression and prognosis of osteosarcoma patients

MicroRNA-106a-5p (miR-106a-5p) functions as a tumor suppressor in osteosarcoma cells. Here, we aimed to identify novel target genes of miR-106a-5p in osteosarcoma, as well as to investigate their prognostic value and the biological functions. At first, the mammalian runt-related factor 1 (RUNX1) was identified as one of the target genes of miR-106a-5p in osteosarcoma cells by luciferase reporter gene assay, real-time quantitative RT-PCR and Western blot analysis. Then, the expression levels of miR-106a-5p and RUNX1 in osteosarcoma tissues were detected, and their associations with clinicopathological features and patients' prognosis were statistically analyzed. Compared with adjacent non-cancerous tissues, miR-106a-5p and RUNX1 mRNA/protein expression in osteosarcoma tissues were significantly decreased and increased, respectively (all P < 0.01). Low miR-106a-5p, high RUNX1 and miR-106a-5p-low/RUNX1-high expression in osteosarcoma tissues were all significantly associated with advanced Enneking stage, positive metastasis and shorter overall survival (all P < 0.05). Moreover, miR-106a-5p and RUNX1 expression, alone or in combination, were identified as independent prognostic factors for osteosarcoma patients' overall survival. Functionally, the enforced expression of miR-106a-5p significantly suppressed proliferation and invasion of osteosarcoma cells, while the overexpression of RUNX1 effectively reversed its suppressive roles. In conclusion, our findings show the dysregulation of miR-106a-5p-RUNX1 axis in human osteosarcoma tissues and suggest its crucial roles in cancer progression and patients' prognosis. More interestingly, miR-106a-5p may function as a tumor suppressor in osteosarcoma cells via regulating its target gene RUNX1.     

血液中microRNA-21对肺癌诊断价值的Meta分析

目的 通过Meta分析已报道文献中的微小RNA-21(microRNA-21,miR-21)对肺癌的诊断价值,探讨血液中的miR-21对肺癌的诊断价值.方法 检索中国知网、维普、万方、PubMed和Embsase等数据库,按照已定的纳入和排出标准筛选文献.采用Quadas量表评估纳入文献质量.通过Stata12和Meta-Disc软件合并纳入文献的诊断值并检测评估异质性和发表偏倚.结果 共检出994篇相关文献,通过筛选,7篇文献(含1 088例样本)纳入本次Meta分析.通过合并文献得出,miR-21对肺癌的诊断灵敏度、特异度、阳性似然比、和阴性似然分别为0.68(95％CI:0.55～0.79),0.82(95％CI:0.76～0.86),3.70(95％CI:2.60～5.25)和0.39(95％CI:0.27～0.58);综合诊断指标综合曲线下面积(SROC)和诊断比值比(DOR)分别为0.84(95％CI:0.80～0.87)和9.90 (95％CI:4.75～20.63).结论 目前研究证据表明,miR-21对肺癌具有较好诊断价值,可作为肺癌诊断的潜在指标.     

急性缺血性脑卒中患者血清微小RNA-590水平变化及意义

目的]探究急性缺血性脑卒中(AIS)患者血清微小RNA-590(miR-590)水平变化及与预后的关系。[方法]选取2019年2月—2020年10月接受静脉溶栓和动脉取栓治疗的176例AIS患者作为研究对象,根据治疗后90天的改良Rankin量表(mRS)评分将其分为预后良好组(n＝123;mRS评分0～2分)、预后不良组(n＝53;mRS评分3～6分)。用实时荧光定量PCR法检测AIS患者血清miR-590水平;用受试者工作特征(ROC)曲线评价miR-590预判AIS预后的价值;用Logistic回归分析AIS预后的风险因素。[结果]预后不良组的年龄、男性占比、糖尿病史占比、空腹血糖(FPG)、总胆固醇(TC)、美国国立卫生研究院卒中量表(NIHSS)评分和入院至溶栓时间(DNT)均高于预后良好组(P＜0.05)。按1∶1进行倾向性评分匹配后,治疗前预后不良组的miR-590相对水平低于预后良好组,预后不良组和预后良好组的治疗后miR-590相对水平均高于治疗前,治疗后预后不良组的miR-590相对水平升高程度高于预后良好组,差异均有统计学意义(P＜0.05)。治疗前miR-590预判AIS预后的ROC曲线下面积为0.793,高于miR-590差值(治疗前和治疗后血清miR-590相对水平的差值)(P＜0.001)。Logistic回归分析结果显示FPG、TC、NIHSS评分和DNT是AIS预后的独立危险因素(P＜0.05),治疗前血清miR-590水平是AIS预后的独立保护因素(P＜0.001)。[结论]AIS患者静脉溶栓和动脉取栓治疗前血清miR-590水平低与AIS预后不良有关。     

过表达microRNA-144通过靶向调节泛素样PHD和环指结构域1基因诱导非小细胞肺癌凋亡

目的探讨microRNA-144 (miR-144)对非小细胞肺癌(Non-small-cell lung cancer,NSCLC)细胞增殖以及预后的影响。方法通过癌症基因组图谱(The cancer genome atlas,TCGA)检测NSCLC患者组织中miR-144以及泛素样PHD和环指结构域包含1(Ubiquitin like PHD and ring finger domain 1,UHRF1)的表达预后以及两者表达的相关性。采用CCK-8、AO/EB以及γH2A检测不同表达miR-144对NSCLC细胞活性、细胞凋亡、增殖以及DNA损伤的影响,Western blot检测PCNA、Bax、Bcl-2及UHRF1的表达。荧光素酶报告证明miR-144以及UHRF1的靶向关系。结果在NSCLC患者组织中,miR-144表达水平较低,低表达miR-144 NSCLC患者生存时间明显低于高表达miR-144者。UHRF1在NSCLC患者组织中明显升高,且在不同年龄、性别、TNM分期和种族有显著差异。低表达UHRF1非小细胞肺癌患者生存率明显高于高表达UHRF1的NSCLC患者。过表达miR-144能够明显降低A549细胞活性,诱导A549细胞凋亡,增加γH2ax表达;抑制PCNA以及Bax蛋白表达,上调Bcl-2表达。荧光素酶报告证明UHRF1是miR-144的靶基因。同时,低表达miR-144能够使UHRF1表达增加,过表达miR-144能够抑制UHRF1的表达。结论过表达miR-144通过靶向UHRF1诱导A549细胞凋亡以及DNA损伤,参与NSCLC的病理生理过程,进而影响预后。