卵巢癌耐药相关微小RNA的研究进展

越来越多的证据表明,微小RNA(miRNA)的异常表达与肿瘤发生发展、转移及预后等密切相关.最近研究发现,异常表达的miRNA与卵巢癌化疗抵抗相关,提示miRNA将可能成为卵巢癌治疗的新靶点.进一步阐明miRNA与卵巢癌耐药的相关机制将有利于miRNA靶向治疗的研究.     

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。     

卵巢上皮恶性肿瘤侵袭转移相关miRNA的筛选与鉴定

目的 探索与卵巢上皮恶性肿瘤侵袭转移可能相关的miRNA.方法 采用miRNA芯片,筛选SKOV-3ip和SKOV-3细胞差异表达miRNA.利用生物信息学软件TargetScan、MicroCosm、PicTar和GO,预测差异表达miRNA的靶基因及其功能.采用实时逆转录聚合酶链反应(real-time RT-PCR)技术,验证与卵巢癌侵袭转移可能相关的5种miRNA(let-7a、let-7e、let-7f、miR-22和miR-886-5p)在SKOV-3ip和SKOV-3细胞的表达.同时,检测这5种miRNA在另外一组侵袭转移能力不同的卵巢癌细胞株HO8910和HO-8910PM中的表达,并进行统计学分析.结果 基因芯片筛选显示,42种miRNA在SKOV-3ip和SKOV-3细胞株表达差异明显.进一步分析显示,let-7a、let-7e、let-7f、miR-22和miR-886-5p等5种miRNA可能与卵巢癌的侵袭转移密切相关.real-time RT-PCR结果证实,let-7f和miR-22在两组侵袭转移能力不同的卵巢癌细胞(SKOV-3和SKOV-3ip细胞、HO-8910和HO-8910PM细胞)表达差异有统计学意义(均P＜0.05).结论 let-7f和miR-22在侵袭转移能力强的卵巢癌细胞中低表达,可能具有抑癌基因的作用.     

miR-296在卵巢癌中的表达及其靶基因的预测研究

目的 观察miR-296在卵巢癌中的表达及其靶基因的预测.方法 收集卵巢癌患者组织样本22例和妇科活检为正常的样本22例,在提取RNA后进行RNA纯度检测,进而检测miR-296在卵巢癌中的表达;在卵巢癌细胞中稳定转入高表达miR-296的质粒,miRNA芯片进行靶基因检测.结果 卵巢癌组织中miR-296高表达,与正常对照组相比,差异有统计学意义(P＜0.001).在卵巢癌细胞系中,当高表达miR-296时,microRNA143和microRNA215在转染组中高表达,microRNA96、microRNA551b、microRNA502-3p低表达.结论 miR-296在卵巢癌中高表达,且其潜在的靶基因有可能作为临床卵巢癌诊断的标志.     

卵巢上皮癌原发耐药相关microRNA的初步研究

目的 探讨难治性卵巢上皮癌耐药相关microRNA(miRNA),并预测其调控的靶基因,为进一步进行卵巢上皮癌耐药相关生物学通路研究提供基础.方法 收集接受满意肿瘤细胞减灭术的100例Ⅱb~Ⅲc期卵巢上皮癌患者的一般资料和组织学标本,根据术后化疗效果将患者分为铂类敏感组(n=91)和难治性卵巢癌组(n=9),每组各选取3例患者使用TaqMan Real-time PCR microRNA Array进行分析,得出差异表达的miRNA.根据靶基因预测网站的分析、miRNA及其靶基因在肿瘤中的研究,筛选出目标miRNA作为区分难治性卵巢癌组与铂类敏感组的标志物以待进一步研究.结果 miRNA表达谱芯片得到两组差异表达的miRNA共118个,其中难治性卵巢癌组与铂类敏感组相比,表达降低的miRNA有42个,表达升高的miRNA有76个.综合现有文献及靶基因预测网站分析,选取6个miRNA作为区分难治性卵巢癌组与铂类敏感组的目标miRNA.与铂类敏感组相比,miRNA-21和miRNA-27a在难治性卵巢癌组中表达明显升高(P﹤0.001),miRNA-100、miRNA-770-5p、miR-NA-200c和miRNA-497在难治性卵巢癌组中表达降低(P﹤0.05).结论 miRNA-21等118个miRNA可能与卵巢上皮癌耐药相关,miRNA-21、miRNA-27a的升高及miRNA-100、miRNA-770-5p、miRNA-200c、miRNA-497的降低可能提示卵巢上皮癌原发耐药.     

微小RNA与卵巢癌

微小RNA (miRNA)的异常表达与卵巢癌的发生发展密切相关,其参与调控卵巢癌的发生、侵袭和转移以及抗癌药物耐药性形成等过程.作为一种重要的生物学标志物,miRNA有望成为卵巢癌早期诊断与预后判断的新靶点,为卵巢癌的治疗提供新的途径.     

基于功能组学数据识别卵巢癌风险microRNA

目的通过生物信息学与免疫组化实验相结合的方法,探讨新的卵巢癌发病相关microRNA(miRNA)。方法选取2004年1月至2006年3月间哈尔滨医科大学附属第一医院及哈尔滨医科大学附属第三医院收治的70例上皮性卵巢癌手术病例石蜡包埋样本,采用SP免疫组化法进行检测,设阴性对照组及阳性对照组,阳性反应观察采用双盲法。以Gene Ontology为功能组学背景,度量不同基因之间的功能相似性得分,根据得分综合排序,寻找卵巢癌风险miRNA。应用该方法对2 588个miRNA进行优化排序,通过免疫组化实验方法对风险miRNA在卵巢癌中调控位点进行证实。结果优化后,卵巢癌相关miRNA如miR-200a/b/c排在前列,排序前10位miRNA中的大部分都在正常组织和卵巢癌组织中呈差异表达。miR-125a调控的人表皮生长因子-2(HER-2)呈高表达趋势。HER-2表达水平与临床分期,病理分级及存活时间显著相关,差异有统计学意义(P<0.05)。结论 miR-125a通过调控HER-2影响卵巢癌的临床病理特征,可成为新的卵巢癌相关风险因子。     

微小RNA与卵巢癌铂类耐药的研究进展

微小RNA(micmRNA,miBNA)是广泛存在于生物体内的一类非编码的小RNA,miRNA是细胞增殖、死亡、应激抗性和脂肪代谢的关键调节因素,体外实验已经证实miRNA在卵巢癌耐药中起重要作用.卵巢癌铂类耐药的产生与发展是十分复杂的过程,研究结果显示,miRNA的表达对卵巢癌细胞株的铂类药物敏感性有着重要作用,能够...     

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.     

经阴道多普勒超声联合血清HE4检测对卵巢癌术后复发的监测价值

目的 探讨阴道多普勒超声联合HE4对卵巢癌术后复发的监测的价值.方法 选取健康体检健康人群、卵巢癌患者和卵巢良性肿瘤患者各60例,对所有受试者行阴道多普勒超声检查以及血清HE4水平检测.60例卵巢癌患者行卵巢癌术后接受联合化疗6～8个疗程,随访过程中监测各项指标以及观察卵巢癌是否复发.结果 血清HE4的表达量和表达阳性率在健康人群、卵巢良性肿瘤以及卵巢癌中差异显著,具有统计学意义(P＜0.05).阴道多普勒超声联合血清HE4对卵巢肿瘤的诊断灵敏度、特异度以及准确度分别为94.9％、93.7％和94.4％,均优于阴道多普勒超声或血清HE4单独检测,差异显著(P＜0.05).卵巢癌术后复发者血清HE4含量平均为(227.14±53.79) pmol·L-1,卵巢癌术后未复发者血清HE4含量平均为(53.77±23.58)pmol·L-1,差异具统计学意义(P＜0.05),多普勒超声联合血清HE4检测对卵巢癌术后复发诊断的灵敏度、特异度以及准确度分别为92.3％、90.5％和91.7％,优于阴道多普勒超声或血清HE4单独检测卵巢癌复发,差异具统计学意义(P＜0.05).结论 经阴道超声多普勒联合血清HE4检测可显著提高对卵巢癌术后复发的敏感性、特异性以及准确性,具重要的临床应用价值.     

Impact of pattern of recurrence on clinical outcome of ovarian cancer patients: clinical considerations

Recurrence of disease represents a clinical challenge in ovarian cancer patients. The aim of this study was to analyse the distribution and pattern of recurrence and their association with clinical outcome in a large series of ovarian cancer patients. This study was conducted on 328 primary untreated ovarian cancer patients. For each relapse, information on date of clinical/instrumental recurrence, and pattern of disease presentation were retrieved. In stage III-IV cases (n = 270), diffuse abdominal carcinomatosis occurred in 62.1% of cases, while recurrences presented as a single lesion or multiple nodules occurred in 9.9% and 26.7% of cases, respectively. Pattern of recurrence as carcinomatosis was shown to be associated with unfavourable outcome even when stratified according to platinum free interval (PFI) duration. In multivariate analysis, pattern of recurrence and PFI duration retained an independent prognostic role for post-relapse survival. Duration of PFI and type of recurrence may independently influence post-relapse survival in ovarian cancer patients.     

Ovarian cancer stem cells and inflammation

Epithelial ovarian cancer (EOC) is the fourth leading cause of cancer-related deaths in women in the United States and the leading cause of gynecologic cancer deaths. The major limiting factor in the treatment of ovarian cancer is recurrence and chemoresistance. Individuals who succumb to advanced-stage ovarian cancer inevitably become refractory to chemotherapy, resulting in disease progression and death. The source of recurrence and lack of response to chemotherapy is unknown. The focus of this review is to evaluate the question of recurrence and chemoresistance based on the concept of the cancer stem cells and inflammation.     

Solitary gastric recurrence from ovarian carcinoma: a case report and literature review

BACKGROUND: Isolated gastric recurrence due to ovarian cancer is a rare event and is usually associated with gastrointestinal symptoms. CASE REPORT: We report a case of an isolated gastric recurrence of ovarian carcinoma in an otherwise asymptomatic 42-year-old woman in whom diagnosis was made using the FDG-PET/CT scan followed by laparoscopy. CONCLUSIONS: In rare cases, ovarian cancer can directly recur on the stomach without any symptoms. FDG-PET/CT scan and diagnostic laparoscopy are effective in leading to an early diagnosis of disease recurrence.     

上皮性卵巢癌复发的影响因素分析

背景与目的:中晚期上皮性卵巢癌复发率较高,生存率低。在没有可靠的早期诊断技术的情况下,了解其复发的影响因素对于预后的判断和指导治疗具有非常重要的意义。本文旨在探讨上皮性卵巢癌复发的影响因素。方法:109例上皮性卵巢癌,对可能导致病人复发的影响因素:年龄,病理类型,临床分期,新辅助化疗,术后化疗方案和化疗疗程,初次手术治疗情况;以及手术残留癌灶大小,用Logistic回归方法进行回顾性总结和分析。结果:109例患者中复发36例,复发率为33.0％,中位复发时间19个月;109例病人总的5年生存率为62.7％,复发病人5年生存率43.3％;盆腔复发病人的5年生存率为50.1％,盆腔外复发病人的5年生存率36.1％,二者相比较无统计学意义(P>0.05)。单因素分析分别显示粘液性腺癌和临床分期为Ⅰ期的复发风险较低(回归系数β=1.565和-1.799,P=0.0120和0.026);而化疗疗程>8个疗程的病人复发风险明显高于1～4疗程和6～8疗程 的病人(回归系数β=-3.591和-1.500,P<0.001和=0.038)。多因素分析显示病理类型、临床分期和术后化疗疗程数是影响卵巢癌治疗后复发的独立危险因素(RR=3.473,4.713和6.140,P<0.05)。结论:临床分期是影响复发的显著因素,早期诊断对于降低卵巢癌复发率十分重要。合理的化疗计划对于卵巢癌的治疗是必要的,     

Modern management of recurrent ovarian carcinoma. A systematic approach to a chronic disease

The management of ovarian cancer entails a complex blend of medical and surgical interventions. Managing patients with recurrent ovarian cancer increases the complexity of therapies and adds palliative interventions. The presence of recurrent ovarian cancer is both emotionally and physically taxing for patients as well as their caregivers. With an increasingly informed patient population, a balance must be achieved between easily accessible information enabling patients to know that they now have an incurable disease and support for their hopes and desires to still overcome their cancer. The decision tree in the management of recurrent ovarian cancer blends many different factors. This discussion will separate those factors as if they are pure elements. We will address management based on response to primary therapy and time to recurrence, the location of recurrence, symptoms of recurrence, the patient's histopathology, and the patient's primary stage as it relates to the extent of disease present at the start of chemotherapy.     

18F-FDG PET/CT在卵巢癌复发和转移上的临床价值

在妇科肿瘤患者中,卵巢癌具有较高的发病率以及复发率。早期发现卵巢癌局部复发或远处转移,对临床治疗方案的制定、患者的生存期和生存质量有重要的意义。与CT和MRI相比,¹⁸F-脱氧葡萄糖(FDG)PET/CT在卵巢癌复发和转移的探测方面具有较高的敏感性和特异性,而且¹⁸F-FDG PET/CT可以早期监测和评估放疗和化疗后的效果,对于卵巢癌的后续治疗意义较大。本文就¹⁸F-FDG PET/CT在卵巢癌复发、转移、评价疗效和预后判断等方面的临床价值作一综述。     

ZNF671 DNA methylation as a molecular predictor for the early recurrence of serous ovarian cancer

Serous ovarian cancer is the most frequent type of epithelial ovarian cancer. Despite the use of surgery and platinum‐based chemotherapy, many patients suffer from recurrence within 6 months, termed platinum resistance. Currently, the lack of relevant molecular biomarkers for the prediction of the early recurrence of serous ovarian cancers is linked to the poor prognosis. To identify an effective biomarker for early recurrence, we analyzed the genome‐wide DNA methylation status characteristic of early recurrence after treatment. The patients in The Cancer Genome Atlas (TCGA) dataset who showed a complete response after the first therapy were categorized into 2 groups: early recurrence serous ovarian cancer (ERS, recurrence ≤12 months, n = 51) and late recurrence serous ovarian cancer (LRS, recurrence >12 months, n = 158). Among the 12 differently methylated probes identified between the 2 groups, we found that ZNF671 was the most significantly methylated gene in the early recurrence group. A validation cohort of 78 serous ovarian cancers showed that patients with ZNF671 DNA methylation had a worse prognosis (P < .05). The multivariate analysis revealed that the methylation status of ZNF671 was an independent factor for predicting the recurrence of serous ovarian cancer patients both in the TCGA dataset and our cohort (P = .049 and P = .021, respectively). Functional analysis revealed that the depletion of ZNF671 expression conferred a more migratory and invasive phenotype to the ovarian cancer cells. Our data indicate that ZNF671 functions as a tumor suppressor in ovarian cancer and that the DNA methylation status of ZNF671 might be an effective biomarker for the recurrence of serous ovarian cancer after platinum‐based adjuvant chemotherapy.     

铂类化疗敏感型卵巢上皮癌复发的影响因素

背景与目的:卵巢癌患者中对铂类化疗敏感者较耐药者预后好,但是铂类化疗敏感型卵巢上皮癌患者中仍有较高的复发率,从而影响此类患者的预后。本研究旨在总结对铂类化疗敏感型卵巢上皮癌患者的临床特点,探讨影响其复发的因素。方法:回顾性分析和总结1993～1999年中山大学肿瘤防治中心收治的90例临床完全缓解超过6个月以上的对铂类化疗敏感型卵巢上皮癌患者复发的影响因素。复发相关的单因素分析采用χ2检验,多因素分析采用Cox模型。结果:90例卵巢上皮癌患者中出现复发者36例,复发率为40.0%,中位复发时间20个月。复发的部位以盆腔最多,占50.0%(18/36)。90例患者总的3年、5年生存率分别为79.6%、69.5%。36例复发患者3年、5年生存率分别为62.3%、39.6%。单因素分析显示铂类化疗敏感型卵巢上皮癌中FIGO分期早、无新辅助化疗、粘液性癌者复发风险低(P=0.001,P=0.002和P=0.025)。经Cox多因素分析显示仅FIGO分期是肿瘤复发的独立危险因子(RR=1.771,P=0.003)。术后采用CBP和铂类其它组合的方案化疗对铂类化疗敏感型卵巢上皮癌复发的影响差异无显著性,过多疗程的术后化疗并不能减少复发。结论:FIGO分期是影响铂类化疗敏感型卵巢上皮癌复发的显著因素,早期诊断对于降低复发十分重要。     

Up-regulation of CD44 in the development of metastasis,recurrence and drug resistance of ovarian cancer

The clinical significance of Cluster of Differentiation 44 (CD44) remains controversial in human ovarian cancer. The aim of this study is to evaluate the clinical significance of CD44 expression by using a unique tissue microarray, and then to determine the biological functions of CD44 in ovarian cancer. In this study, a unique ovarian cancer tissue microarray (TMA) was constructed with paired primary, metastatic, and recurrent tumor tissues from 26 individual patients. CD44 expression in TMA was assessed by immunohistochemistry. Both the metastatic and recurrent ovarian cancer tissues expressed higher level of CD44 than the patient-matched primary tumor. A significant association has been shown between CD44 expression and both the disease free survival and overall survival. A strong increase of CD44 was found in the tumor recurrence of mouse model. Finally, when CD44 was knocked down, proliferation, migration/invasion activity, and spheroid formation were significantly suppressed, while drug sensitivity was enhanced. Thus, up-regulation of CD44 represents a crucial event in the development of metastasis, recurrence, and drug resistance to current treatments in ovarian cancer. Developing strategies to target CD44 may prevent metastasis, recurrence, and drug resistance in ovarian cancer.     

The role of bevacizumab in recurrent,platinum-sensitive ovarian cancer

The majority of women with ovarian cancer will experience a recurrence of their disease despite aggressive primary cytoreduction and adjuvant cytotoxic chemotherapy. Notwithstanding the high rate of recurrence, targeted and biologic agents have helped to decrease the dependence on cytotoxic chemotherapy. Bevacizumab, a vascular endothelial growth factor inhibitor, has been shown to cause regression in tumor vasculature, inhibition of angiogenesis and prevention of progenitor cell recruitment. Phase III clinical trials of bevacizumab in patients with primary epithelial ovarian cancer and in patients with platinum-sensitive ovarian cancer have shown an improvement in progression free survival without an appreciable difference in overall survival. The addition of bevacizumab to standard cytotoxic chemotherapy regimens has demonstrated improved response rates, and improved progression free survival. These results have stimulated research in additional angiogenesis inhibitors and trials to further incorporate bevacizumab into the treatment schema for patients with recurrent ovarian cancer.