血清外泌体miRNAs在晚期胃癌患者化疗疗效预测中的价值

  目的  通过筛选晚期胃癌患者血清外泌体miRNAs，探讨外泌体miRNAs在化疗疗效预测中的价值。  方法  收集苏州大学附属第一医院2018年4月至2020年11月确诊为初治Ⅳ期胃腺癌患者36例病例资料，均接受不少于2个周期的Xelox或SOX方案化疗，根据实体肿瘤疗效评价标准（RECIST标准）分为敏感组和耐药组，收集患者首次化疗前的血清标本，提取血清外泌体RNA，行miRNAs测序，筛选出差异表达显著的miRNAs，并采用实时荧光定量PCR对筛选的miRNAs进行验证，使用受试者工作特征曲线（receiver operating characteristic curve，ROC），判断验证后的外泌体miRNAs在疗效预测中的价值。  结果  36例Ⅳ期胃癌患者按照RECIST标准，分为敏感组15例，耐药组21例。两组患者血清外泌体中共筛选出527个差异表达的miRNAs，经qRT-PCR验证后，仅3个外泌体miRNAs差异具有统计学意义，ROC曲线预测miRNA-106a-5p、miRNA-1323、miRNA-202-3p与胃癌化疗疗效的曲线下面积分别为0.952、0.949、0.946，灵敏度分别为90.5%、90.1%、85.7%，特异度分别为:93.3%、86.7%、93.1%。  结论  检测晚期胃癌患者血清外泌体miRNA-106a-5p、miRNA-1323、miRNA-202-3p，可能对化疗疗效具有一定的预测价值。      

胃癌患者血清miR-26a/b的表达水平及诊断价值

  目的  探讨胃癌患者血清miR-26a/b诊断胃癌的价值及其与临床病理特征的关系。  方法  采用qRT-PCR法检测121例胃癌患者和116例正常人(对照组)血清miR-26a/b的表达量,分析miR-26a/b与胃癌临床病理特征的关系,构建受试者工作特征曲线(ROC),分析miR-26a/b诊断胃癌的价值。  结果  胃癌患者血清miR-26a和miR-26b相对表达水平均低于对照组(1.60±1.02 vs.5.35±0.44;1.44±0.71 vs.5.35±0.71;P < 0.05),miR-26a/b的表达水平与患者TNM分期相关,且miR-26a的相对含量还与浸润深度、有无淋巴结转移相关(P < 0.05)、性别、分化程度、组织学类型等其他临床病理特征无显著相关(P>0.05);miR-26a的ROC曲线下面积为0.828(95% CI:0.776~0.881),灵敏度为73.3%,特异度为81.0%;miR-26b的ROC曲线下面积为0.853(95%CI:0.801~0.906),灵敏度为68.1%,特异度为99.2%。  结论  miR-26a/b在胃癌患者血清中表达显著降低,并且其表达水平与胃癌的临床分期、浸润深度及有无淋巴结转移相关,有着较高的诊断价值。      

血清外泌体miR-148a联合miR-106a检测用于胃癌筛查的研究探讨

  目的  寻找合适的外泌体miRNA作为肿瘤生物标志物，辅助临床进行胃癌筛查。  方法  将miR-148a及miR-106a作为潜在标志物，研究两种miRNA在癌组织及癌旁组织表达的差异性。选取2017年9月至2018年9月在福建省立医院诊治的初诊胃癌术后标本共37对（胃癌组织及癌旁组织）进行组织学miRNA检测。选取初诊胃癌患者83例为胃癌组，良性病变的患者78例为对照组，全部进行血清外泌体miRNA检测。  结果  与癌旁组织相比，癌组织中miR-148a表达水平降低，miR-106a表达水平升高，联合检测2-△△CP[△CP=CP_{（miR-148a）}-CP_{（miR-106a）}]值降低。与对照组相比，胃癌患者中血清外泌体miR-106a表达水平降低，联合检测2-△△CP值升高。血清外泌体联合检测2-△△CP值对胃癌组和对照组鉴别的曲线下面积[AUC为0.844（95%CI:0.782~0.905，P < 0.001）]，大于miR-148a及miR-106a单独检测，其cut-off值为0.315 3，敏感度为91，6%，特异度为64.1%。  结论  血清外泌体miR-148a联合miR-106a检测的2-△△CP值可以作为胃癌筛查的手段。      

胃癌发生过程中风险miRNAs筛选及其诊断效能评价的多中心研究

目的 探索胃癌发生过程的风险miRNAs,为上消化道机会性筛查中早期胃癌识别提供依据。方法 纳入2021年6月至2023年8月在广西医科大学附属肿瘤医院、右江民族医学院附属医院、桂林市人民医院3个中心进行上消化道癌机会性筛查的人群。选取健康体检者107例、早期胃癌患者71例、进展期胃癌患者97例。首先采用转录组测序筛选差异表达miRNAs,然后在3组前瞻性人群的血浆样本中通过RT-qPCR验证差异表达的miRNAs,最后采用受试者工作特征（receiver operating characteristic,ROC）曲线评估miRNAs的诊断效能。结果 转录组测序的差异基因分析共筛选出胃癌发生过程中的6个差异表达miRNAs,包括miR-3176、miR-885-5p、miR-203a-3p、miR-452-5p、miR-223-3p、miR-219a-2-3p。RT-qPCR结果显示,miR-452-5p在早期胃癌及进展期胃癌患者中表达上调（均P<0.001）。ROC曲线显示,miR-452-5p诊断早期胃癌和进展期胃癌的曲线下面积（area under the curve,AU...     

循环血清 miR -17 -92 簇: 一种新的胃癌诊断标志物

目的：探索胃癌患者血清miR-17-92簇的表达水平,评价其对胃癌诊断和检测的临床应用价值.方法：qRT-PCR法检测79名胃癌患者及38名正常对照组中血清miR-17-92簇的表达水平.结果：miR-NAs可以在血清中稳定存在.血清miR-17-3p、miR-17-5p、miR-18a-5p、miR-19a-3p、miR-19b-3p、miR-20a-5p、miR-92a-3p的表达 水平在胃癌患者中均高于正常对照组（P=0.005、0.009、0.006、0.003、0.045、0.016）.miR-17-3p 在七个miRNA中的ROC曲线下面积最大,且诊断的特异度高达89.00%.miR-17-3p和miR-19a-3p 联合检测对胃癌诊断特异度有明显提 高,高达94.70%,灵敏度为63.16%.miR-17-92簇的表达与胃癌的分期及淋巴结转移有关（P＜0.01、=0.02）.结论：血清miR-17-92簇可以作为一种潜在的胃癌生物标志物,辅助胃癌诊断.     

透明细胞肾细胞癌外泌体miR-133a表达及临床意义

目的 探讨外泌体微小RNA(miR)-133a在透明细胞肾细胞癌(ccRCC)中的表达及临床意义。方法 收集2015年9月至2019年1月在我院行根治性肾切除术的293例ccRCC患者的血清样本，另外收集200例健康体检者的血清样本作为对照组。提取血清及组织外泌体总RNA,采用TaqMan低密度芯片(TLDA)鉴定差异表达的miRNA,并通过实时荧光定量PCR(qPCR)进行验证。随访患者的总生存期(OS)和无病生存期(DFS)。结果 与健康对照组血清和癌旁正常组织比较，TLDA筛选得到3个在ccRCC患者血清外泌体和肿瘤组织中差异表达的miRNAs(miR-133a、miR-210、miR-7-1)。qPCR证实外泌体miR-133a表达在ccRCC患者中显著下调(P<0.001)。ccRCC患者外泌体miR-133a表达与肿瘤组织表达呈正相关(r=0.467,P<0.001)。受试者工作特征(ROC)曲线证实血清外泌体miR-133a诊断ccRCC曲线下面积(AUC)为0.78(95%CI:0.747～0.729)。术前血清外泌体miR-133a表达与TNM分期、Fuh...     

血清miR-6861-5p检测在乳腺癌临床诊治中的价值探讨

  目的  研究miR-6861-5p在乳腺癌患者血清中的表达,并探讨miR-6861-5p表达在乳腺癌患者临床诊治中的价值。  方法  通过实时荧光定量PCR法（RT-qPCR）检测2012年1月至2015年6月山东省德州市第二人民医院112例乳腺癌患者、37例乳腺良性病变和53例健康女性血清miR-6861-5p相对表达量,分析血清miR-6861-5p表达与乳腺癌患者临床病理和术后复发之间的关系,并探讨其表达对乳腺癌的临床诊断价值。  结果  乳腺癌患者血清miR-6861-5p相对表达量为（7.99±1.63）,显著高于良性病变患者（6.45±1.06）（P＜0.05）和健康研究对象（6.43±1.28）（P＜0.05）;血清miR-6861-5p表达与乳腺癌患者淋巴结转移、肿瘤组织ER、PR和HER-2表达、TNM分期、分子分型以及组织学分期显著相关（均P＜0.05）;肿瘤切除后乳腺癌患者血清miR-6861-5p表达量显著降低;治疗前血清miR-6861-5p诊断乳腺癌的敏感度和特异度分别为69.6%和77.8%,且与乳腺癌患者术后复发率有关。  结论  miR-6861-5p在乳腺癌患者血清中表达上调,是术前乳腺癌诊断、肿瘤分期、癌细胞转移和术后监测乳腺癌复发的潜在生物标志物。      

MicroRNA-100在胃癌患者血清中的表达及临床意义

  目的  探讨MicroRNA-100（miR-100）在胃癌患者血清中的表达及临床意义。  方法  选择蚌埠医学院第一附属医院手术及随访资料完整的胃癌患者（40例）和健康对照者（40例）为研究对象,提取血清总miRNA,在建立了稳定、敏感的血清miR-100绝对定量检测方法（qRT-PCR）的基础上,检测胃癌和健康对照者血清中miR-100表达水平。分析胃癌和健康对照者血清中miR-100表达差异及血清中miR-100表达水平与胃癌临床病理参数的关系。  结果  胃癌患者血清中miR-100的表达水平为（2.78±1.92）fmol/L,显著高于健康对照者[（0.19±0.15）fmol/L,P < 0.01],同时miR-100的受试者工作特征曲线显示,血清miR-100对胃癌诊断具有良好的特异度和敏感度（曲线下面积0.985）;进一步分析发现:血清中miR-100的表达与性别、年龄、肿瘤直径大小、浸润深度、分化程度、淋巴结转移数量、TNM分期等无明显差异（P > 0.05）。  结论  血清miR-100的检测可能有助于胃癌的诊断。      

胃癌患者血清外泌体粒径的临床意义

目的 探讨胃癌患者血清外泌体粒径的变化及其在胃癌患者预后评估方面的临床意义。方法 收集2021年5月至2022年2月皖南医学院收治的胃癌病例，同时收集同期健康体检者作为对照组。采用PEG沉淀法提取32例胃癌患者及26例健康志愿者的血清外泌体，运用纳米颗粒跟踪技术(NTA)检测胃癌血清外泌体的粒径和浓度；分析血清外泌体粒径大小与胃癌临床病理特征的关系；采用受试者工作特征曲线(ROC)分析血清外泌体粒径和浓度检测对胃癌分期的预测价值。结果 胃癌患者血清外泌体的浓度为(9.53±6.24)×10⁷particals/ml,高于健康对照组的(6.14±3.43)×10⁷particals/ml,差异有统计学意义(P<0.05);且胃癌患者血清外泌体粒径为(108.35±16.63)nm,显著小于健康对照组的(128.23±20.24)nm,差异有统计学意义(P<0.001)。血清外泌体粒径与胃癌患者的分期相关，即随着胃癌分期的增加，血清外泌体的粒径显著下降(P<0.05);ROC曲线结果显示，血清外泌体粒径诊断胃癌的灵敏度、特异度分...     

肝细胞肝癌患者血清microRNA-224水平变化及其临床诊断意义的研究

  目的  探讨肝细胞肝癌患者血清microRNA-224水平变化及其临床诊断意义。  方法  采用实时荧光定量PCR检测42例肝细胞肝癌患者（hepatocellular carcinoma,HCC）、36例肝硬化患者（LC）、55例慢性乙型肝炎患者（CHB）以及40例健康体检者（NC）血清miR-224表达量。计算miR-224相对表达量。通过分析受试者工作特征曲线（ROC）判断miR-224表达水平在肝癌诊断中的灵敏度和特异度。  结果  HCC组患者血清miR-224相对表达量均高于CHB组、LC组和NC组,差异有统计学差异（P < 0.05或P < 0.01）。HCC患者血清miR-224相对表达量与甲胎蛋白（AFP）呈正相关（P < 0.05）。与肿瘤大小、TNM分期、肿瘤分化程度以及淋巴结转移无相关性（P>0.05）。ROC分析确定了最佳的miR-224相对表达量的临界值为3.47,灵敏度被确定为82.2%,特异性为92.8%,曲线下面积（AUC）为0.935。  结论  肝细胞肝癌患者血清中miR-224特异性高表达,将使其可能成为一种新的血清学指标用于肝细胞肝癌的诊断。      

Circulating exosomal miR-125a-5p as a novel biomarker for cervical cancer

Exosomal microRNAs (miRs/miRNAs) have been reported to be associated with cervical cancer. The aim of the present study was to investigate circulating exosomal miRNA as a biomarker for cervical cancer diagnosis. In the present study, samples from 6 patients with cervical cancer and 6 healthy control subjects were retrieved for exosomal RNA-sequencing. The results revealed that a total of 39 miRNAs were differentially expressed between patients with cervical cancer and healthy controls (P<0.001; fold-change >2.0). Exosomal miR-125a-5p was further quantified in plasma from 60 subjects, which included 22 healthy individuals and 38 patients with cervical cancer. miR-16a-5p served as the reference miRNA for quantitative PCR analysis of exosomal miR-125a-5p in patients with cervical cancer and healthy individuals. The results revealed that exosomal miR-125a-5p expression levels in the patients with cervical cancer were significantly lower than those in the healthy controls (P<0.001). Receiver operating characteristic (ROC) curve analyses were performed and the results revealed that the level of plasma exosomal miR-125a-5p was a potential marker for differentiating between non-cervical cancer and cervical cancer, with an ROC area under the curve of 0.7129. At the cut-off value of 2.537 for miR-125a-5p, cervical cancer diagnostic sensitivities and specificities were 59.1 and 84.2%, respectively. The present study provides confirmation that exosomal miR-125a-5p could potentially serve as a biomarker for cervical cancer diagnosis. The present study involved only a small number of clinical samples; more samples are required to support the conclusions of the present study.     

胃癌细胞侵犯骨髓患者临床实验室检查特点分析

  目的  分析胃癌骨髓侵犯患者临床实验室检测结果的特点，筛选对胃癌细胞骨髓侵犯具有提示意义的实验室指标。  方法  回顾性分析2013年1月至2021年3月河北医科大学第四医院收治的30例胃癌发生骨髓侵犯患者，收集血常规、凝血功能、免疫、生化及骨髓等临床指标检测结果资料。分析上述患者与未发生骨髓侵犯的Ⅳ期胃癌患者临床实验室检测结果的差异，并制作受试者工作特征（receiver operating characteristic，ROC）曲线评价各指标在提示胃癌细胞骨髓侵犯中的意义。  结果  与未发生骨髓侵犯的Ⅳ期胃癌患者相比较，发生骨髓侵犯的患者血小板（Plt）计数、凝血酶原时间（PT）、凝血酶时间（TT）、乳酸脱氢酶（LDH）、D二聚体（D-DIMER）、纤维蛋白原降解产物（FDP）、CEA及CA72-4等指标的检测结果差异均具有统计学意义（均P<0.05）。其中FDP的ROC曲线下面积最大（AUC=0.988），且阳性似然比最高。同时，两组患者外周血幼红、幼粒细胞检出率差异具有统计学意义（P<0.01）。  结论  胃癌骨髓侵犯患者的部分临床实验室检测指标较未发生骨髓侵犯患者异常检出率更高且变化更为明显，上述指标对骨髓侵犯具有提示意义。      

非小细胞肺癌吉非替尼耐药相关miRNAs的筛选鉴定

  目的  miRNA是一类通过结合mRNA调节基因表达的非编码单链小分子RNA,本研究目的是探讨非小细胞肺癌（NSCLC）中miRNA与吉非替尼耐药的关系。  方法  CCK8法检测NSCLC吉非替尼耐药细胞PC9/GR相对于亲本细胞PC9的耐药倍数;miRNA芯片检测PC9/GR与PC9中miRNA的表达差异;RT-PCR验证miRNA芯片结果。将差异表达的miRNA模拟物/抑制剂转染至PC9/GR中,观察其对吉非替尼敏感性的影响。  结果  吉非替尼对PC9和PC9/GR的IC₅₀值分别为42.89 nmoL/L和3.87 μ moL/L,耐药倍数为90.23倍。miRNA芯片结果显示,PC9/GR与PC9比较55条有差异表达miRNAs（P < 0.01）,其中在PC9/GR上调的miRNAs有21条,包括miRNA-1 246、miRNA-125b等;下调的miRNAs有34条,包括miRNA-224、miRNA-125a~5p等。RT-PCR进一步验证其中9条miRNAs,有8条与芯片结果趋势一致。将上述8条miRNAs的模拟物/抑制剂转染至PC9/GR中,发现miRNA-125a~5p模拟物可降低吉非替尼敏感性。  结论  PC9/GR与PC9的miRNA表达存在差异,miRNA可能与NSCLC吉非替尼耐药相关,miRNA-125a~5p可促进PC9/GR对吉非替尼产生耐药。      

Pap Smear miR-92a-5p and miR-155-5p as potential diagnostic biomarkers of squamous intraepithelial cervical cancer

Background: one of the female-specific diseases with a high incidence and mortality is cervical cancer. The main cause of cervical cancer is infection with Human papilloma virus (HPV). Low-grade squamous intraepithelial lesions (LSIL) and High-grade squamous intraepithelial lesions (HSIL) usually is caused by an HPV infection. Considering the role of microRNAs (miRNAs) as diagnostic biomarkers for a variety of cancers, the aim of this study was to determine miR-92a-5p and miR-155-5p expression levels in LSIL and HSIL Pap Smear samples. Methods: After initial bioinformatic studies, A total of 75 samples (25 samples of patients with LSIL, 25 patients with HSIL and 25 healthy individuals) were subjected to RNA extraction and cDNA synthesis. The expressions levels of confirmed miRNAs in samples of patients with LSIL, HSIL and healthy individuals were evaluated by Real time PCR analysis. To demonstration the role of predicted miRNAs as novel biomarkers in diagnosis of LSIL and HSIL, ROC curve analysis was done. Results: Bioinformatics results showed that miR-92a-5p and miR-155-5p target the HPV E6 and E7 genes. The expression levels of these miRNAs were strikingly higher in Pap smear of patients with LSIL than in the healthy individuals (35.36, P = 0.001) (62.23, P = 0.001). Similarity, expression levels of miR-92a-5p and miR-155-5p were amazingly higher in patients with HSIL than in the healthy individuals (33.62, P= 0.001) (69.07, P= 0.001). Although, the levels of miR-92a-5p (0.95, P = 0. 85) and miR-155-5p (1.11, P = 0.84) exhibited no statistical differences between patients with LSIL and HSIL. Also, ROC curve analyses verified that miR-92a-5p and miR-155-5p are specific and sensitive and may serve as new biomarkers for the early detection of cervical cancer. Conclusion: These data suggest miR-92a-5p and miR-155-5p, which are upregulated in LSIL and HSIL, can be consider as predictive biomarkers for the prognosis of cervical cancer patients.     

长链非编码RNA FAM201A靶向miR-488-3p对胃癌细胞生物学行为及放射敏感性的影响

  目的  探讨长链非编码RNA（lncRNA）序列相似性家族201-成员A（family with sequence similarity 201-member A，FAM201A）靶向miR-488-3p对胃癌细胞生物学行为及放射敏感性的影响。  方法  选取2014年1月至2017年1月间于莱阳中心医院确诊并进行胃癌根治性切除手术（术前均未经过放疗和化疗治疗）的胃癌患者的肿瘤组织标本和配对的非癌性黏膜标本63例，采用qRT-PCR检测63例胃癌组织和与其对应的癌旁组织中FAM201A及miR-488-3p的表达水平。构建抑制FAM201A表达的MGC803胃癌细胞株。四甲基偶氮唑蓝（methyl thiazolyl tetrazolium，MTT）法检测细胞增殖活力，流式细胞术检测细胞凋亡，Transwell法检测细胞的迁移和侵袭能力。用不同辐射强度（0、2、4、6和8 Gy）射线照射抑制FAM201A表达的MGC803细胞，克隆形成实验检测细胞存活分数，绘制单击多靶模型拟合曲线。双荧光素酶报告基因实验和qRT-PCR验证FAM201A和miR-488-3p的靶向关系。  结果  与癌旁组织相比，胃癌组织中FAM201A的表达水平显著升高，miR-488-3p的表达水平显著降低。抑制FAM201A可显著抑制MGC803细胞增殖、迁移和侵袭，促进其凋亡，增加MGC803细胞的放射敏感性。FAM201A可靶向负性调控miR-488-3p表达。抑制miR-488-3p能够逆转抑制FAM201A对细胞MGC803增殖、凋亡、迁移侵袭和放射敏感性的影响。  结论  抑制lncRNA FAM201A通过靶向miR-488-3p可抑制胃癌细胞增殖、迁移和侵袭，促进胃癌细胞凋亡，并增加其放射敏感性。FAM201A有望成为胃癌的新型分子靶点。      

Plasma Circulating Mirnas Profiling for Identification of Potential Breast Cancer Early Detection Biomarkers

Objective: This study aimed to characterize the miRNA expression profiles from plasma samples of our local breast cancer patients in comparison to healthy control by using miRNA PCR Array. Methods: In this study, plasma miRNA profiles from eight early-stage breast cancer patients and nine age-matched (± 2 years) healthy controls were characterized by miRNA array-based approach, followed by differential gene expression analysis, Independent T-test and construction of Receiver Operating Characteristic (ROC) curve to determine the capability of the assays to discriminate between breast cancer and the healthy control. Results: Based on the 372-miRNAs microarray profiling, a set of 40 differential miRNAs was extracted regarding to the fold change value at 2 and above. We further sub grouped 40 miRNAs of breast cancer patients that were significantly expressed at 2-fold change and higher. In this set, we discovered that 24 miRNAs were significantly upregulated and 16 miRNAs were significantly downregulated in breast cancer patients, as compared to the miRNA expression of healthy subjects. ROC curve analysis revealed that seven miRNAs (miR-125b-5p, miR-142-3p, miR-145-5p, miR-193a-5p, miR-27b-3p, miR-22-5p and miR-423-5p) had area under curve (AUC) value > 0.7 (AUC p-value < 0.05). Overlapping findings from differential gene expression analysis, ROC analysis, and Independent T-Test resulted in three miRNAs (miR-27b-3p, miR-22-5p, miR-145-5p). Cohen’s effect size for these three miRNAs was large with d value are more than 0.95. Conclusion: miR-27b-3p, miR-22-5p, miR-145-5p could be potential biomarkers to distinguish breast cancer patients from healthy controls. A validation study for these three miRNAs in an external set of samples is ongoing.