血清microRNA 在多形性胶质母细胞瘤术预后评估中的研究

目的:筛选多形性胶质母细胞瘤（glioblastoma multiform,GBM）患者术前、术后血清中差异表达的microRNAs（miRNAs）,并探讨差异表达的miRNAs与患者术后预后的相关性。方法:收集2006年1 月至2009年6 月48例北京天坛医院经临床病理诊断为GBM患者的术前术后血清样本。采用Solexa 测序的方法初步筛选出术前术后表达量有差异的 miRNA ,用实时荧光定量PCR（quantitative real-time PCR ,RT-qPCR）的方法对每个样本进行逐一验证,应用t 检验的方法筛选出满足条件的miRNA（两组之间的平均值差异在2 倍以上,且P < 0.05）,对48例患者进行随访,统计生存时间,根据48例患者中位生存时间494 d,将所有标本分为长生存期组和短生存期组,应用Kaplan-Meier 法和Log-rank 检验,研究患者术后血清miRNAs的表达量与患者生存时间之间是否存在统计学意义的相关性。结果:Solexa 结果显示,有63个miRNA 表达量存在差异,基于本研究先前的研究成果和其他文献的报道,从中选出4 个miRNA（miR-26b,miR-30e ,miR-129- 3p,miR-206）进行逐一验证并进行统计学分析,结果只有1 个miRNAs（miR-30e）在术后患者血清中的表达水平有明显上调现象（术前与术后表达水平平均值差异≥ 2 倍且P < 0.05）,随访结果显示,生存时间> 494 d,患者术后血清miR-30e 的表达水平有降低的趋势（P < 0.05）,但生存分析显示,患者术后血清中miR-30e 的表达量与患者总生存时间之间差异无统计学意义（P = 0.101）。 结论:GBM患者术前术后血清中差异表达的miRNA 只有miR-30e ,且术后患者血清中的miR-30e 水平与肿瘤负荷成负相关关系。生存分析结果显示,术后患者血清miR-30e 的表达水平与患者的预后没有明显的相关性。      

卵巢癌血清相关miRNAs的筛选及其临床意义

背景与目的：卵巢癌预后较差,发现时通常是晚期,需找寻与卵巢癌发生、发展相关的诊治方法。该研究检测miRNA在上皮性卵巢癌患者术前外周血清及良性卵巢肿瘤患者外周血清中表达情况的差异,筛选差异有统计学意义的miRNA并分析其与上皮性卵巢癌患者临床病理特征、预后等的关系。方法：定制研究相关的48种miRNA表达谱芯片,通过TaqMan低密度微阵列芯片,筛选出有统计学意义的miRNA。采用实时荧光定量聚合酶链反应(real-time fluorescent quantitative polymerase chain reaction,RTFQ-PCR)法验证筛选的miRNA在卵巢良、恶性肿瘤患者血清中的表达情况,选择具有统计学意义的miRNA行大样本验证并分析其与肿瘤分期、组织病理及预后等的关系。结果：通过TaqMan低密度微阵列芯片筛选和RTFQ-PCR验证,发现miR-125b在上皮性卵巢癌患者血清中的表达高于良性肿瘤患者(P=0.039),miR-125b在早期患者中的表达量高于晚期患者(P=0.003),术后无残余肿瘤患者表达量高于术后有残余肿瘤患者(P=0.013)。血清miR-125b高表达有利于卵巢癌患者无进展生存期(progression-free survival,PFS)延长(P=0.003),但对总生存期(overall survival,OS)无明显影响(P=0.069)。结论：miR-125b在上皮性卵巢癌的发生、发展中起着关键作用,与患者预后相关,是预测卵巢癌复发的潜在基因,但在肿瘤不同期别的表达情况发生变化,在早期作用比较明显,在晚期或肿瘤残余较多的患者表达较不明显,其作用机制有待进一步研究。     

miR-10a-5p基因甲基化对卵巢癌铂类耐药的影响

目的 卵巢癌是死亡率最高的妇科肿瘤,较强的化疗耐药性是其预后差的主要原因之一,为了阐明卵巢癌对铂类药物的耐药机制,本研究探讨miRNA基因的甲基化水平对卵巢癌铂类耐药的影响.方法 将卵巢癌组织分为敏感组和耐药组,每组各3例;采用基因芯片技术,对比分析了两组微RNA(microRNA,miRNA)的表达差异;采用实时荧光定量PCR,分别在6例敏感和3例耐药组织、铂类药物敏感(CoC1)和耐药的卵巢癌细胞系(CoC1/DDP),检测了候选miRNA的表达差异;应用Massarray技术,检测敏感组织(15例)与耐药组织(6例)中miRNA基因启动子的甲基化差异;应用生物信息学分析,鉴定目标miRNA的潜在靶基因.结果 以铂类药物敏感的卵巢癌组织样本为对照,利用基因芯片筛选,鉴定了6条在耐药组织样本中出现表达上调的miRNA(miR-493-3p、miR-10a-5 p、miR-16-2-3p、miR-1248、miR-451a、miR-628-3p)和6条表达下调的miRNA(miR-509-3p、miR-1197、miR-376a-3p、miR-1273a、miR-550a-3p、miR-19b-3p).组织验证发现,miR-509-3p、miR-493-3p、miR-10a-5p、miR-16-2-3p和miR-451a,与芯片结果一致;培养细胞研究发现,4条miRNA的表达调控方式与组织芯片结果一致,miR-10a-5p、miR-16-2-3p、miR-1248和miR-628-3p在耐药细胞系中高表达.进一步研究发现,与敏感肿瘤组织相比,耐药组织中miR-10a-5p基因启动子的甲基化水平出现显著降低,P=0.04.结合生物信息学预测HOXA1和USF2为miR-10a-5p与耐药相关的靶基因.结论 与敏感组相比,耐药组miR-10a-5p基因启动子甲基化水平显著降低,miR-10a-5p表达升高,通过抑制 HOXA1和USF2,抑制细胞凋亡,导致铂类化疗药物耐受.     

Circulating Cell-free miRNA Expression and its Association with Clinicopathologic Features in Inflammatory and Non- Inflammatory Breast Cancer

Recent discovery showing the presence of microRNAs (miRNAs) in the circulation sparked interest in their use as potential biomarkers. Our previous studies showed the diagnostic potential of miR-451 as a serological marker for inflammatory breast cancer (IBC), miR-337- 5p and miR-30b for non-inflammatory breast cancer (non-IBC). The aim of this study is to investigate the prognostic values of circulating miRNAs by comparing the amounts of 12 circulating miRNAs in the serum of IBC and non-IBC from Tunisian breast cancer patients, and by determinating whether correlated pairs of miRNAs could provide useful information in the diagnosis of IBC and non-IBC patients. TaqMan qPCR was performed to detect circulating expression of miRNAs in serum of 20 IBC, 20 non-IBC and 20 healthy controls. Nonparametric rank Spearman rho correlation coefficient was used to examine the prognostic value of miRNAs and to assess the correlation profile between miRNAs expression. Further, a large number of miRNAs were highly correlated (rho>0.5) in both patients groups and controls. Also, the correlations profiles were different between IBC, non-IBC and healthy controls indicating important changes in molecular pathways in cancer cells. Our results showed that miR-335 was significantly overexpressed in premenopausal non-IBC patients; miR-24 was significantly overexpressed in non-IBC postmenopausal patients. Patients with previous parity had higher serum of miR-342-5p levels than those without. Furthermore, patients with HER2 IBC present lower serum levels of miR-15a than patients with HER2- disease. Together, these results underline the potential of miRNAs to function as diagnostic and prognostic markers for IBC and non-IBC, with links to the menopausal state, Her2 status and parity.     

miR-203a-3p 在胰腺癌组织与细胞中的表达及其对 BxPC-3 细胞增殖、 迁移和侵袭的影响

目的：探讨miR-203a-3p对胰腺癌BxPC-3细胞增殖、迁移和侵袭能力的影响。方法：运用癌症基因组图谱（TCGA） 数据库筛选胰腺癌组织和癌旁组织中差异表达的miRNA,分析miRNA高表达与低表达时胰腺癌患者的生存率和临床分期;利用 TarBase数据库分析miRNA与癌症相关的GO功能与KEGG通路,利用DIANA Tools、miRDB和TargetScan网站预测miR-203a-3p 的靶基因。将miR-203a-3p mimic及NC mimic、miR-203a-3p inhibitor及NC inhibitor转染至BxPC-3细胞,用qPCR法检测胰腺癌 细胞和胰腺正常上皮细胞HPNE中miR-203a-3p、miR-192-5p和miR-451a表达水平,以CCK-8法、Transwell小室法和克隆形成实 验分别检测BxPC-3细胞的增殖、迁移、侵袭和集落形成能力。结果：通过TCGA数据库筛选出18个胰腺癌组织中差异表达的 miRNA,其中miR-203a-3p、miR-192-5p、miR-451a具有物种保守性 ,且其与胰腺癌临床癌症分期、细胞周期和患者生存率相 关（均P<0.05）;生物信息学网站预测显示miR-203a-3p的候选靶基因是PPM1A,PPM1A与多基因存在相互作用。miR-203a-3p、 miR-192-5p和miR-451a在BxPC-3和Aspc-1细胞中均高表达（均P<0.01）。miR-203a-3p mimic组BxPC-3细胞中miR-203a-3p表 达水平以及细胞增殖、迁移和侵袭能力均显著提高（均P<0.01）：miR-203a-3p inhibitor组细胞中miR-203a-3p表达水平以及细胞 增殖、迁移和侵袭能力均显著降低（均P<0.01）。结论：miR-203a-3p在胰腺癌组织及细胞中均高表达,其表达与患者生存和临床 分期相关,可调控BxPC-3细胞的增殖、迁移和侵袭能力。     

Identification of 9 serum microRNAs as potential noninvasive biomarkers of human astrocytoma

BackgroundCirculating microRNAs (miRNAs) are emerging as promising biomarkers for human cancer. In the current study, we investigated the potential use of serum miRNAs as biomarkers for diagnosis and prognosis in a cohort of Chinese astrocytoma patients.MethodsAn initial screening of the circulating miRNA expression profile was performed on pooled serum samples from 10 preoperative patients and 10 healthy controls using a TaqMan low-density array. The selected serum miRNAs were then validated in 90 preoperative patients and 110 healthy controls who were randomly divided into a training set and a validation set. An additional double-blind test was performed in 50 astrocytomas and 50 controls to assess the serum miRNA-based biomarker accuracy in predicting astrocytoma. The differentially expressed miRNAs were evaluated in paired preoperative and postoperative serum samples from 73 astrocytoma patients. The correlation of the miRNA levels with survival in astrocytoma samples was estimated.ResultsNine serum miRNAs were significantly increased in the astrocytoma patients. The biomarker composed of these 9 miRNAs had high sensitivity, specificity, and accuracy. These 9 miRNAs were markedly decreased in the serum after operation. The upregulation of miR-20a-5p, miR-106a-5p, and miR-181b-5p was associated with advanced clinical stages of astrocytoma. Kaplan-Meier survival analysis showed that the high expression of miR-19a-3p, miR-106a-5p, and miR-181b-5p was significantly associated with poor patient survival. Finally, the combined 3-miRNAs panel was an important prognostic predictor, independent of other clinicopathological factors.ConclusionsThe results indicated the potential of serum miRNAs as novel diagnostic and prognostic biomarkers for human astrocytoma.     

Association of miR-193b Down-regulation and miR-196a up-Regulation with Clinicopathological Features and Prognosis in Gastric Cancer

Dysregulated expression of microRNAs (miRNAs) has been shown to be closely associated with tumordevelopment, progression, and carcinogenesis. However, their clinical implications for gastric cancer remainelusive. To investigate the hypothesis that genome-wide alternations of miRNAs differentiate gastric cancer tissuesfrom those matched adjacent non-tumor tissues (ANTTs), miRNA arrays were employed to examine miRNAexpression profiles for the 5-pair discovery stage, and the quantitative real-time polymerase chain reaction (qRTPCR)was applied to validate candidate miRNAs for 48-pair validation stage. Furthermore, the relationshipbetween altered miRNA and clinicopathological features and prognosis of gastric cancer was explored. Amonga total of 1,146 miRNAs analyzed, 16 miRNAs were found to be significantly different expressed in tissues fromgastric cancer compared to ANTTs (p<0.05). qRT-PCR further confirmed the variation in expression of miR-193band miR-196a in the validation stage. Down-expression of miR-193b was significantly correlated with Laurentype, differentiation, UICC stage, invasion, and metastasis of gastric cancer (p<0.05), while over-expression ofmiR-196a was significantly associated with poor differentiation (p=0.022). Moreover, binary logistic regressionanalysis demonstrated that the UICC stage was a significant risk factor for down-expression of miR-193b (adjustedOR=8.69; 95%CI=1.06-56.91; p=0.043). Additionally, Kaplan-Meier survival curves indicated that patientswith a high fold-change of down-regulated miR-193b had a significantly shorter survival time (n=19; mediansurvival=29 months) compared to patients with a low fold-change of down-regulated miR-193b (n=29; mediansurvival=54 months) (p=0.001). Overall survival time of patients with a low fold-change of up-regulated miR-196a (n=27; median survival=52 months) was significantly longer than that of patients with a high fold-changeof up-regulated miR-196a (n=21; median survival=46 months) (p=0.003). Hence, miR-193b and miR-196a maybe applied as novel and promising prognostic markers in gastric cancer.     

Circulating miRNA expression over the course of colorectal cancer treatment

Colorectal cancer (CRC) is the third-most common cancer type in males and the second-most common cancer type in females, and has the second-highest overall mortality rate worldwide. Approximately 50% of patients in stage I–III develop metastases, mostly localized to the liver. All physiological conditions occurring in the organism are also reflected in the levels of circulating microRNAs (miRNAs/miRs) in patients. miRNAs are a class of small, non-coding, single-stranded RNAs consisting of 18–25 nucleotides, which have important roles in various cellular processes. The aim of the present study was to evaluate a panel of seven circulating miRNAs (miR-106a-5p, miR-210-5p, miR-155-5p, miR-21-5p, miR-103a-3p, miR-191-5p and miR-16-5p) as biomarkers for monitoring patients undergoing adjuvant treatment of CRC. Total RNA was extracted from the plasma of patients with CRC prior to surgery, in the early post-operative period (n=60) and 3 months after surgery (n=14). The levels of the selected circulating miRNAs were measured with the miRCURY LNA miRNA PCR system and fold changes were calculated using the standard ∆∆Cq method. DIANA-miRPath analysis was used to evaluate the role of significantly deregulated miRNAs. The results indicated significant upregulation of miR-155-5p, miR-21-5p and miR-191-5p, and downregulation of miR-16-5p directly after the surgery. In paired follow-up samples, the most significant upregulation was detected for miR-106a-5p and miR-16-5p, and the most significant downregulation was for miR-21-5p. Pathway analysis outlined the role of the differentially expressed miRNAs in cancer development, but the same pathways are also involved in wound healing and regeneration of intestinal epithelium. It may be suggested that these processes should also be considered in studies investigating sensitive and easily detectable circulating biomarkers for recurrence in patients.     

结肠癌患者血清中miR-30a、miR-106b的表达及其临床意义

目的:探讨结肠癌患者血清miR-30a、miR-106b的表达及其在结肠癌进展及预测患者预后中的价值。方法:选取2012年06月至2013年02月本院收治的接受结肠癌手术治疗的患者80例为研究对象,20例同期健康人群作为对照组。逆转录 PCR法检测外周血血清中 miR-30a、miR-106b的表达水平,分析患者血清miR-30a、miR-106b表达与结肠癌临床病理及预后的关系。Spearman分析miR-30a、miR-106b表达的相关性。采用多因素logistic回归模型对结肠癌患者的预后影响因素进行分析,采用Kaplan-Meier生存分析miR-30a低表达组、miR-30a高表达组,miR-106b低表达组、miR-106b高表达组患者中位生存时间。结果:结肠癌患者血清miR-30a表达水平（1.03±0.02）低于正常人群（5.15±0.06）,差异具有统计学意义（t=16.38,P=0.01）;结肠癌患者血清miR-106b表达水平（2.62±0.35）高于正常人群（1.15±0.06）,差异具有统计学意义（t=10.17,P=0.03）。miR-30a、miR-106b表达与患者的组织分化类型、浆膜侵犯、淋巴结转移、远处转移、TNM分期有显著相关性(P＜0.05)。多因素logistic回归模型对结肠癌患者预后影响因素进行分析,TNM分期（Ⅲ+Ⅳ）、miR-106b高表达及miR-30a低表达是结肠癌患者预后不良的危险因素。miR-30a低表达组患者总体生存时间（55.3±4.1）个月低于miR-30a高表达组（76.4±2.4）个月（P=0.000）;miR-106b低表达组患者的总体生存时间（75.1±7.3）个月高于miR-106b高表达组（51.8±3.1）个月（P=0.000）。结论:miR-30a在结肠癌患者血清中低表达,miR-106b在结肠癌患者血清中高表达,二者的表达与肿瘤的进展、预后不良有关。     

Identification of Differentially Expressed Mirna by Next Generation Sequencing in Locally Advanced Breast Cancer Patients of South Indian Origin

Purpose: miRNAs are known to be aberrantly expressed in the serum, tissue, and Peripheral Blood Mononuclear Cells (PBMC) of cancer patients and could serve as potential noninvasive diagnostic markers for breast cancer. The aim of this study was to identify the differentially expressed miRNA using next-generation sequencing (NGS) from the paired PBMC samples from breast cancer patients and age-matched healthy individuals and explore their functional significance. Methods: In this study, PBMCs were employed for the detection of miRNAs by NGS in locally advanced breast cancer (LABC) women of South Indian origin who were divided into three age groups, (a) 40yrs-50yrs (b) 50yrs-60yrs and (c) 60yrs-70yrs, compared with age-matched control groups. Results: Four miRNAs (hsa-miR-192-5p, hsa-miR-24-2-2p, hsa-miR-3609, and hsa-miR-664b-3p) were found to be differentially expressed among LABC patients compared with age matched healthy women of the South Indian population. While miR-24-2-5p, miR3609, and miR-664b-3p were down-regulated, miR-192-5p was up-regulated. Gene Ontology (GO) annotations implicated miRNA with signaling pathways in peripheral nerve synapses, glutamatergic synapse, and cell morphogenesis, all of which play a pivotal role in the manifestation of cancer. Conclusion: Four miRNAs- 3 (While miR-24-2-5p, miR3609, and miR-664b-3p) downregulated and one upregulated (miR-192-5p) were identified as potential biomarkers for patients with locally advanced breast cancer. These markers could be validated in studies with a larger sample size.     

Circulating microRNAs from the miR-106a–363 cluster on chromosome X as novel diagnostic biomarkers for breast cancer

Purpose

Novel noninvasive biomarkers with high sensitivity and specificity for the diagnosis of breast cancer (BC) are urgently needed in clinics. The aim of this study was to explore whether miRNAs from the miR-106a–363 cluster can be detected in the circulation of BC patients and whether these miRNAs can serve as potential diagnostic biomarkers.

Methods

The expression of 12 miRNAs from the miR-106a–363 cluster was evaluated using qRT-PCR in 400 plasma samples (from 200 BC patients and 200 healthy controls (HCs)) and 406 serum samples (from 204 BC patients and 202 HCs) via a three-phase study. The identified miRNAs were further examined in tissues (32 paired breast tissues), plasma exosomes (from 32 BC patients and 32 HCs), and serum exosomes (from 32 BC patients and 32 HCs).

Results

Upregulated levels of four plasma miRNAs (miR-106a-3p, miR-106a-5p, miR-20b-5p, and miR-92a-2-5p) and four serum miRNAs (miR-106a-5p, miR-19b-3p, miR-20b-5p, and miR-92a-3p) were identified and validated in BC. A plasma 4-miRNA panel and a serum 4-miRNA panel were constructed to discriminate BC patients from HCs. The areas under the receiver-operating characteristic curves of the plasma panel were 0.880, 0.902, and 0.858, and those of the serum panel were 0.910, 0.974, and 0.949 for the training, testing, and external validation phases, respectively. Two overlapping miRNAs (miR-106a-5p and miR-20b-5p) were consistently upregulated in BC tissues. Except for the expression of the plasma-derived exosomal miR-20b-5p, the expression patterns of exosomal miRNAs were concordant between plasma and serum, indicating the potential use of exosomal miRNAs as biomarkers.

Conclusion

We identified four plasma miRNAs and four serum miRNAs from the miR-106a–363 cluster as promising novel biomarkers for the diagnosis of BC.

     

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。      

Identification of serum miRNAs as novel non-invasive biomarkers for detection of high risk for early gastric cancer

Background:

Many micro-RNAs (miRNAs) are differentially expressed in Helicobacter pylori-infected gastric mucosa and in gastric cancer tissue and previous reports have suggested the possibility of serum miRNAs as complementary tumour markers. The aim of the study was to investigate serum miRNAs and pepsinogen levels in individuals at high risk for gastric cancer both before and after H. pylori eradication.

Methods:

Patients with recent history of endoscopic resection for early gastric cancer and the sex- and age-matched controls were enrolled. Serum was collected from subjects before or after eradication and total RNA was extracted to analyse serum levels of 24 miRNAs. Serum pepsinogen (PG) I and II levels were measured using enzyme-linked immunosorbent assay kits.

Results:

Using miR-16 as an endogenous control, the relative levels of miR-106 and let-7d before and after H. pylori eradication and miR-21 after eradication were significantly higher in the high-risk group than in the controls. H. pylori eradication significantly decreased miR-106b levels and increased let-7d only in the control group. After eradication, the combination MiR-106b with miR-21 was superior to serum pepsinogen and the most valuable biomarker for the differentiating high-risk group from controls.

Conclusion:

Serum miR-106b and miR-21 may provide a novel and stable marker of increased risk for early gastric cancer after H. pylori eradication.     

Down Regulated Expression Levels of miR-27b and miR-451a as a Potential Biomarker for Triple Negative Breast Cancer Patients Undergoing TAC Chemotherapy

Background: Triple negative breast cancer (TNBC) is associated with poor prognosis, aggressive phenotype(s) of tumours, partial chemotherapy response, and lack of clinically proven therapies. MicroRNAs (miRNAs) can target and modulate key genes that are involved in TNBC chemotherapy. Deregulated miRNA expression is highly involved in anti-cancer drug resistance phenotype and thus, miRNAs tend to be promising candidates for prediction of chemotherapy response and recurrence. Aim: This study aimed to investigate the expression levels of selected miRNAs (miR-21, miR-27b, miR-34a, miR-182, miR-200c and miR-451a) in cancerous and normal adjacent tissues of TNBC patients and to correlate with the clinicopathological data. Methods: Forty-one (41) FFPE tissue block of histopathologically confirmed TNBC patients was collected. Total RNA from the cancerous and adjacent non-cancerous tissues were isolated, transcribed, and pre-amplified. The relative expression level of miRNAs in tumour and normal adjacent tissues of TNBC patients was analysed using qRT-PCR. Results: Out of six miRNAs studied, the relative expression of miR-27b and miR-451a were found to be significantly lower in cancerous as compared to normal adjacent tissues of TNBC patients. In addition, a significant down regulation of miR-451a was also observed in infiltrating ductal carcinoma subtype, stages I and II, in both grade II and III, premenopausal and postmenopausal as well as in those with positive axillary lymph node metastases. Conclusion: The results suggest the possible utilization of miR-27b and miR-451a expression levels as potential predictive risk markers for TNBC patients undergoing TAC chemotherapy.     

微RNA-106b～25基因簇在肿瘤中的研究进展

微RNA(miRNA)-106b～25基因簇由miR-106b、miR-93及miR-25组成,与miR-17～92原癌基因簇有着极高的同源性.已有的研究表明,该簇miRNA成员在多种肿瘤中高表达.这些miR通过抑制细胞周期抑制因子p21、p57及凋亡抑制因子Bim来促进肿瘤细胞增殖,抑制细胞凋亡.在转化生长因子β(T...     

人脑胶质瘤中miR-106b~25基因簇表达的研究

  目的  检测胶质瘤细胞系及组织中miR-106b~25基因簇成员miR-106b、miR-93、miR-25的表达情况。  方法  运用实时定量PCR的方法检测不同人脑胶质母细胞瘤细胞系及不同病理级别胶质瘤标本中miR-106b~25基因簇成员miR-106b、miR-93和miR-25的表达水平。原位杂交技术检测含不同病理级别及瘤旁正常脑组织的胶质瘤组织芯片中miR-106b~25基因簇成员的表达情况,进而分析该簇miRNAs的表达与胶质瘤病理级别的相关性。  结果  以正常脑组织表达量为基准,3种miRNA在所有检测细胞系中均有增高的趋势。收集43例胶质瘤标本中,RT-PCR结果显示,随着肿瘤病理级别的升高,3种miRNA在各组中的平均表达量也逐步升高。其中,miR-106b（F=4.479,P=0.018）和miR-93（F=3.493,P=0.040）各组间的表达差异均有统计学意义。而miR-25表达无明显的统计学差异（F=2.766,P=0.075）。原位杂交结果显示3种miRNA的表达在高级别胶质瘤中的表达量明显高于低级别肿瘤。Spearman等级相关分析表明3种miRNA的表达信号强度分布均与胶质瘤的WHO病理分级呈正相关,在miR-106b、-93、-25中的相关系数分别为r_s=0.617（P＜0.001）、r_s=0.438（P＜0.001）、r_s=0.463（P＜40.001）。  结论  miR-106b~25在胶质瘤中表达呈不同程度增高,并与肿瘤分级呈正相关。      

Analysis of miR-205 and miR-155 expression in the blood of breast cancer patients

The purpose of this study was to identify and validate circulating microRNAs (miRNAs) in human plasma for use as breast cancer (BC) biomarkers and to analyze their relationship to clinicopathologic features and its preliminary biological function. Genome-wide expression profiling of miRNAs in BC was investigated by microarray analysis. miR-155 was up-regulated greater than two-fold in BC compared with Normal Adjacent Tissue (NAT), whereas let-7b, miR-381, miR-10b, miR-125a-5p, miR-335, miR-205 and miR-145 were down- regulated greater than two-fold. Our hypothesis was that circulating miRNAs are also present and differentially expressed in the serum of BC patients compared to controls. Using real-time PCR (RT-PCR), we analyzed miR-205 and miR-155 in archived serum from 30 participants, 20 with breast cancer and 10 healthy people. miR-205 was down-regulated in BC patient serum while miR-155 was up-regulated. Furthermore, we analyzed the relationship between the expression levels of these two miRNAs and the clinicopathologic parameters of BC patients. High expression of miR155 was associated with clinical stage, molecular type, Ki-67 and p53 in BC patients (P<0.05). By contrast, we found no significant correlation between miR-205 and BC patient clinicopathologic parameters. Functional analysis showed that ectopic expression of miR-205 significantly inhibits cell proliferation and promotes apoptosis. miR-205 was down-regulated and miR-155 was up-regulated in BC patient serum. miR-155 was positive correlated with clinical stage and ki-67 and negatively correlated with p53 status.